(775 days)
The DiaSorin Molecular Simplexa™ Congenital CMV Direct is a real-time PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of cytomegalovirus (CMV) from saliva swabs and urine from infants less than 21 days of age. Positive results from saliva are presumptive and should be confirmed with urine. The results of the Simplexa™ Congenital CMV Direct assay should be used in conjunction with the results of other clinical findings as an aid in the diagnosis of congenital CMV infection.
This test has not been cleared for screening of blood products for the presence of CMV or for use with samples other than urine and saliva swabs.
DiaSorin Molecular's Simplexa™ Congenital CMV Positive Control Pack is intended to be used as a control with the Simplexa Congenital CMV Direct kit for use on the LIAISON MDX instrument. This control is not intended for use with other assays or systems.
The Simplexa™ Congenital CMV Direct assay is a real-time PCR system that enables the direct amplification and detection of CMV DNA from either saliva swab or urine specimens without nucleic acid extraction. The system consists of the Simplexa™ Congenital CMV Direct Reaction Mix, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories.
In the Simplexa™ Congenital CMV Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify CMV DNA. A well-conserved region of the CMV UL83 gene is targeted to identify CMV DNA. An internal control is used to detect PCR failure and/or inhibition.
Here's a breakdown of the acceptance criteria and the study proving the device meets them, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state quantitative acceptance criteria in a dedicated table format. However, the reported performance metrics imply the criteria for acceptance. For the purpose of this response, I'll infer the implicit acceptance criteria based on the demonstrated performance, generally implying "high agreement" for positive and negative cases.
| Metric (Implicit Acceptance Criteria) | Saliva Swab Performance (Retrospective) | Urine Performance (Retrospective) | Saliva Swab Performance (Prospective) | Urine Performance (Prospective) |
|---|---|---|---|---|
| Positive Percent Agreement (PPA) | 100.0% (95% CI: 93% - 100%) | 100.0% (95% CI: 93% - 100%) | 94.1% (95% CI: 73% - 99%) | 95.3% (95% CI: 85% - 99%) |
| Negative Percent Agreement (NPA) | 100.0% (95% CI: 97% - 100%) | 98.4% (95% CI: 94% - 100%) | 99.9% (95% CI: 100% - 100%) | 100.0% (95% CI: 100% - 100%) |
| Reproducibility (%CV) | Between 0.5% and 1.6% | Between 0.7% and 1.5% | N/A (not directly from this study) | N/A (not directly from this study) |
| Analytical Sensitivity (LoD) | 500 Copies/mL (AD-169, Towne, Merlin) in UTM | 400 Copies/mL (AD-169), 800 Copies/mL (Towne), 6400 IU/mL (Merlin) | N/A | N/A |
| Cross-Reactivity | 100% agreement (no cross-reactivity) | 100% agreement (no cross-reactivity) | N/A | N/A |
| Interference | 100% agreement (no interference) | 100% agreement (no interference) | N/A | N/A |
| Microbial Inhibition | 100% agreement (no inhibition) | 100% agreement (no inhibition) | N/A | N/A |
2. Sample Size and Data Provenance
-
Retrospective Study:
- Saliva Swab: 173 total specimens (3 removed due to indeterminate CRM result, 170 analyzed)
- Urine: 173 total specimens
- Provenance: "collected during the clinical study", "stored at a central site", then distributed to three (3) laboratories. The document doesn't explicitly state the country of origin for these retrospective samples, though the testing sites were in the USA. These were pre-selected positive and negative samples based on routine laboratory results.
-
Prospective Study:
- Saliva Swab: 1,859 initially collected, 6 deemed ineligible, resulting in 1,853 analyzed specimens.
- Urine: 1,656 initially collected, 32 deemed ineligible, resulting in 1,624 analyzed specimens.
- Provenance: Prospectively collected (frozen and/or fresh) from ten (10) collection sites across the USA and two (2) collection sites outside the USA. Testing was performed at six (6) testing sites located in the USA.
3. Number of Experts and Qualifications
The document states that a "Composite Reference Method (CRM)" was used, which involved "two (2) validated PCR followed by bi-directional sequencing assays." One (1) central laboratory performed these comparator assays.
- The document does not specify the number of experts used to establish the ground truth or their specific qualifications (e.g., radiologist with 10 years of experience). It relies on the validation of the PCR and bi-directional sequencing assays as the basis for ground truth, implying that these are established and reliable laboratory methods.
4. Adjudication Method
The ground truth for the clinical agreement studies (both retrospective and prospective) was established via a "Composite Reference Method (CRM)".
This CRM "utilized two (2) validated PCR followed by bi-directional sequencing assays. A sample had a final sequencing result of 'Detected' if one or both sequencing results were 'Detected'. Conversely a sample had a final sequencing result of 'Not Detected' if both results were 'Not Detected'." This implies a form of 2+0 or 1+1 adjudication model where if either reference method detects CMV, the sample is considered positive, and both must be negative for the sample to be considered negative.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No, an MRMC comparative effectiveness study was not done. This study focuses on the diagnostic performance of the device itself against a laboratory-based reference method, not on how human readers/clinicians improve with AI assistance.
6. Standalone (Algorithm Only) Performance
Yes, a standalone performance evaluation was done. The Simplexa™ Congenital CMV Direct assay is a real-time PCR assay and its performance was evaluated directly against the Composite Reference Method (CRM) without human-in-the-loop assistance. The reported PPA and NPA values represent the algorithm's standalone performance.
7. Type of Ground Truth Used
The ground truth used was expert consensus on laboratory results, specifically based on a "Composite Reference Method (CRM) utilized two (2) validated PCR followed by bi-directional sequencing assays." This is a highly robust and objective form of ground truth for nucleic acid detection devices, often considered a gold standard in molecular diagnostics.
8. Sample Size for the Training Set
The document does not provide information on the sample size used for the training set. This is typical for submissions of this nature, where the focus is on the validation of the final device/algorithm using a separate, independent test set, rather than details of the developmental (training) phase.
9. How the Ground Truth for the Training Set Was Established
The document does not provide information on how the ground truth for the training set was established, as details about the training phase are not included in this summary.
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Image /page/0/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo includes the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text.
November 5, 2022
DiaSorin Molecular LLC Sharon Young Principal Regulatory Affairs Specialist 11331 Valley View Street Cypress, California 90630
Re: K202755
Trade/Device Name: Simplexa Congenital CMV Direct and Simplexa Congenital CMV Positive Control Pack Regulation Number: 21 CFR 866.3181 Regulation Name: Cytomegalovirus Nucleic Acid Detection Device For Congenital Cytomegalovirus Infection Regulatory Class: Class II Product Code: ODZ Dated: September 17, 2020 Received: September 21, 2020
Dear Sharon Young:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal
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statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Maria I. Garcia -S
Maria Garcia, Ph.D. Assistant Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K202755
Device Name
Simplexa™ Congenital CMV Direct and Simplexa™ Congenital CMV Positive Control Pack
Indications for Use (Describe)
Simplexa™ Congenital CMV Direct Catalog Number MOL2250
The DiaSorin Molecular Simplexa™ Congenital CMV Direct is a real-time PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of cytomegalovirus (CMV) from saliva swabs and urine from infants less than 21 days of age. Positive results from saliva are presumptive and should be confirmed with urine. The results of the Simplexa™ Congenital CMV Direct assay should be used in conjunction with the results of other clinical findings as an aid in the diagnosis of congenital CMV infection.
This test has not been cleared for screening of blood products for the presence of CMV or for use with samples other than urine and saliva swabs
Simplexa™ Congenital CMV Positive Control Pack Catalog Number MOL2260 DiaSorin Molecular's Simplexa™ Congental CMV Positive Control Pack is intended to be used as a control with the Simplexa Congenital CMV Direct kit for use on the LIAISON MDX instrument. This control is not intended for use with other assays or systems.
| Type of Use (Select one or both, as applicable) |
|---|
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) |
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
| CONTINUE ON A SEPARATE PAGE IF NEEDED. |
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510(k) Summarv
Simplexa™ Congenital CMV Direct MOL2250 Simplexa ™ Congenital CMV Positive Control Pack. MOL2260 Nov 2. 2022
Page 1 of 17
| Applicant | DiaSorin Molecular LLC.11331 Valley View StreetCypress, California 90630USA |
|---|---|
| EstablishmentRegistration No. | 2023365 |
| Contact Person | Tara Viviani, RACSr. Directory Molecular Requlatory AffairsTel. 562.240.6680Tara. Viviani@DiaSorin.com |
| Summary Date | November 2, 2022 |
| Proprietary Name | Simplexa™ Congenital CMV Direct andSimplexa™ Congenital CMV Positive Control Pack |
| US ProductCodes/Names andRegulation Numbers | QDZ - Cytomegalovirus nucleic acid detection device for congenitalcytomegalovirus infection 21 CFR § 866.3281OOI - Instrumentation for clinical multiplex test systems 21 CFR § 862.2570 |
| Classification | Class II |
| Predicate Devices | Alethia® CMV Assay Test System DEN180040 (for saliva swab). Currently,there is not a predicate device for urine as a compatible specimen type. |
Intended Use
Simplexa™ Congenital CMV Direct REF MOL2255
The DiaSorin Molecular Simplexa™ Congenital CMV Direct is a real-time PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of cytomegalovirus (CMV) from saliva swabs and urine from infants less than 21 days of age. Positive results from saliva are presumptive and should be confirmed with urine. The Simplexa™ Congenital CMV Direct assay should be used in conjunction with the results of other clinical findings as an aid in the diagnosis of congenital CMV infection.
This test has not been cleared for screening of blood or blood products for the presence of CMV or for use with samples other than urine and saliva swabs.
Simplexa™ Congenital CMVPositive Control Pack REF MOL2265
DiaSorin Molecular's Simplexa™ Congenital CMV Positive Control Pack is intended to be used as a control with the Simplexa™ Congenital CMV Direct kit for use on the LIAISON® MDX instrument. This control is not intended for use with other assays or systems.
Device Description
The Simplexa™ Congenital CMV Direct assay is a real-time PCR system that enables the direct amplification and detection of CMV DNA from either saliva swab or urine specimens without nucleic acid extraction. The system consists of the Simplexa™ Congenital CMV Direct Reaction Mix, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories.
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Simplexa™ Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022 Page 2 of 17
In the Simplexa™ Congenital CMV Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify CMV DNA. A well-conserved region of the CMV UL83 gene is targeted to identify CMV DNA. An internal control is used to detect PCR failure and/or inhibition.
Simplexa™ Congenital CMV Direct REF MOL2255
| Component Name | REF | EC Symbol on Label | Abbreviated Name | Cap Color | Number of Vials | Reactions per Vial/Kit | Volume per Vial | |
|---|---|---|---|---|---|---|---|---|
| Simplexa™ Congenital CMV Direct Reaction Mix | MOL2256 | REAG | C | RM | White | 24 | 1/24 | 50 µL |
Simplexa™ Congenital CMV Direct Components and Descriptions
| Kit Component | Contents | ||||
|---|---|---|---|---|---|
| DNA polymerase, buffer, dNTPs, template DNA (Internal Control), dye-labeled fluorescent probesand primers specific for detection of CMV and for the DNA Internal Control. | |||||
| Simplexa™Congenital CMVDirect ReactionMix (RM) | Target | ProbeFluorophore(Dye) | Excitation (nm) | Emission (nm) | Targeted Gene |
| CMV | FAM | 495 | 520 | UL83 gene | |
| InternalControl RNA(IC) | Q670 | 644 | 670 | N/A | |
| Simplexa™Congenital CMVDirect BarcodeCard | Assay specific parameters and lot information |
Simplexa™ Congenital CMV Positive Control Pack REF MOL2265 Component and Description
| Component Name | REF | Description | CapColor | Numberof Vial | ReactionsperVial/Kits | Volumeper Vial |
|---|---|---|---|---|---|---|
| Simplexa™Congenital CMVDirect PositiveControl | MOL2256 | Inactivated CMV | Red | 10 | 1/10 | 50µL |
Materials Supplied Separately
Direct Amplification Disc Kit (REF MOL1455) Direct Amplification Discs for use on the LIAISON® MDX
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510(K) Summary Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022 Page 3 of 17
Comparison to Predicate Device
| Comparison toPredicate Device | Predicate Device:Alethia CMV Assay Test System(DEN180040) | Candidate Device:Simplexa™ Congenital CMV Direct andSimplexa™ Congenital CMV PositiveControl Pack |
|---|---|---|
| Product Code | QDZ | Same |
| RegulationNumber andDescription | 21 CFR § 866.3381 – Cytomegalovirusnucleic acid detection device for congenitalcytomegalovirus infection. | Same |
| OrganismDetected | cytomegalovirus | Same |
| Measurand | gene | A well-conserved region of the CMV UL83 |
| Intended Use Kit | The Alethia CMV Assay Test Systemincludes separately provided test kits for theAlethia CMV DNA Amplification Assay andthe Alethia CMV External Control Reagents.The Alethia CMV DNA Amplification Assay,performed on the Alethia instrument, is aqualitative, in vitro diagnostic test system forthe direct detection of Cytomegalovirus(CMV) DNA in saliva samples from neonatesyounger than 21 days of age. The test is usedas an aid in the diagnosis of congenital CMVinfection. The results of this test should beused in conjunction with the results of otherclinical findings.Flocked swabs should be used to collectsaliva from neonates. The swab can becollected dry, without viral transport media(VTM), or placed in no more than 1 mL VTM.The Alethia CMV External Control Reagentsare used as part of a routine quality controlprogram to aid the user in detection ofunexpected conditions that may lead to testerrors. The external controls are intended foruse with the Alethia CMV DNA AmplificationAssay; the controls are not intended for usewith other assays or systems. | The DiaSorin Molecular Simplexa™Congenital CMV Direct is a real-time PCRintended for use on the LIAISON MDXinstrument for the in vitro qualitative detectionof cytomegalovirus (CMV) from saliva swabsand urine from infants less than 21 days ofage. The Simplexa™ Congenital CMV Directis an aid in the diagnosis of congenital CMVinfection. |
| Intended UseControl Pack | The Alethia CMV Assay Test Systemincludes separately provided test kits for theAlethia CMV DNA Amplification Assay andthe Alethia CMV External Control Reagents.The Alethia CMV External Control Reagentsare used as part of a routine quality controlprogram to aid the user in detection ofunexpected conditions that may lead to testerrors. The external controls are intended foruse with the Alethia CMV DNA AmplificationAssay; the controls are not intended for usewith other assays or systems. | The Simplexa™ Congenital CMV PositiveControl Pack is intended to be used as acontrol with the Simplexa™ Congenital CMVDirect kit. This control is not intended for usewith other assays or systems. |
| Comparison toPredicate Device | Predicate Device:Alethia CMV Assay Test System(DEN180040) | Candidate Device:Simplexa™ Congenital CMV Direct andSimplexa™ Congenital CMV PositiveControl Pack |
| AutomatedSystem (Sampleto Answer) | Yes | Same |
| Instrumentation | Alethia™ Instrument; Meridian Bioscience,Inc. | LIAISON® MDX |
| SampleTypes/MediaType | Dry flocked saliva swab or saliva swab in 1 mLVTM from infants < 21 days old. | Saliva swab in BD Universal Viral Transport(UVT), Copan UTM® (1mL or 3mL), RemelM4RT®, M4®, and M6® transport and urinefrom infants less than 21 days old. |
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Simplexa™ Congenital CMV Direct Simplexa™ Congenital CMV Positive Control Pack. M Nov 2. 2022
Page 4 of 17
CLINICAL AGREEMENT
The performance of the Simplexa™ Congenital CMV Direct assay was established in a clinical study that included two (2) cohorts based on sample status. Specifically, prospective (pre-selected positive and negative samples based on routine laboratory results) samples from infants less than twentyone (21) days of age, were tested in the clinical agreement study.
Retrospective Study
A total of 346 retrospective specimens were collected during the clinical study. Specimens were enrolled, aliquoted and shipped to a central site where they were distributed for Simplexa™ Congenital CMV Direct testing at three (3) laboratories. One (1) central laboratory performed both comparator PCR/bi-directional sequencing assays for the two (2) part Composite Reference Method (CRM). The Composite Reference Method (CRM) utilized two (2) validated PCR followed by bi-directional sequencing assays. A sample had a final sequencing result of 'Detected' if one or both sequencing results were 'Detected'. Conversely a sample had a final sequencing result of 'Not Detected' if both results were 'Not Detected'.
Of the 346 specimens tested, 170 results were generated for saliva swab specimens and 173 results were generated for urine specimens. The results are presented in Tables 3a and 3b.
The saliva swab study included 173 specimens in the analysis. Three (3) specimens were removed from analysis. The three (3) samples yielded an indeterminate final result with the Composite Reference Method (CRM) CMV-2 PCR/Bi-directional sequencing assay preventing the algorithm from producing a result for use as a comparator.
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510(k) Summarv
Simplexa™ Congenital CMV Direct MOL2250 Simplexa ™ Congenital CMV Positive Control Pack. MOL2260 Nov 2. 2022
Page 5 of 17
Table 3a. Simplexa™ Congenital CMV Direct Clinical Agreement - Saliva Swab (Retrospective)
| Clinical Agreement | PPA | NPA | |||
|---|---|---|---|---|---|
| Simplexa™ CongenitalCMV Direct Results | Composite Reference Method (CRM) | ||||
| Detected | Not Detected | Total | |||
| Detected | 53 | 0 | 53 | 100.0%(53/53)95% CI:93% - 100% | 100.0%(117/117)95% CI: 97% - 100% |
| Not Detected | 0 | 117 | 117 | ||
| Total | 53 | 117 | 170 |
PPA = Positive Percent Agreement, NPA = Negative Percent. 95% Cl = 95% Confidence Interval
The 95% confidence intervals (CI) were calculated following the Wilson Score method
Table 3b. Simplexa™ Congenital CMV Direct Clinical Agreement - Urine (Retrospective)
| Clinical Agreement | PPA | NPA | |||
|---|---|---|---|---|---|
| Simplexa™ CongenitalCMV Direct Results | Detected | Not Detected | Total | ||
| Detected | 49 | 2a | 51 | 100.0%(49/49)95% CI:93% - 100% | 98.4%(122/124)95% CI: 94%- 100% |
| Not Detected | 0 | 122 | 122 | ||
| Total | 49 | 124 | 173 |
a Two (2) urine samples were positive by routine methodology.
PPA = Positive Percent Agreement, NPA = Negative Percent Agreement. 95% Cl = 95% Confidence Interval
The 95% confidence intervals (CI) were calculated following the Wilson Score method.
Prospective Study
A total of one thousand eight hundred fifty-nine (1,859) saliva swab specimens and/or one thousand six hundred fifty-six (1,656) urine specimens were prospectively collected as frozen and/or fresh specimens. Of these collected specimens, six (6) saliva swab and thirty-two (32) urine specimens were deemed ineligible and removed from analysis. Specimens were collected from ten (10) collection sites across the USA and two (2) collection sites outside the USA. Testing was performed at six (6) testing sites located in the USA. One (1) central laboratory performed both comparator PCR/bi-directional sequencing assays for the two (2) part Composite Reference Method (CRM).
The Composite Reference Method (CRM) utilized two (2) validated PCR followed by bi-directional sequencing assays. A sample had a final sequencing result of 'Detected' if one or both sequencing results were 'Detected'. Conversely a sample had a final sequencing result of 'Not Detected' if both results were 'Not Detected'.
Prospective clinical agreement was based on a total of one thousand eight hundred fifty-three (1,853) saliva swab specimens and one thousand six hundred twenty-four (1,624) urine specimens. The results are presented in Tables 4a and 4b.
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510(k) Summarv
Simplexa™ Congenital CMV Direct MOL2250 Simplexa ™ Congenital CMV Positive Control Pack. MOL2260
Nov 2. 2022
Table 4a. Simplexa™ Congenital CMV Direct Clinical Agreement - Saliva Swab (Prospective)
| Clinical Agreement | PPA | NPA | |||
|---|---|---|---|---|---|
| Simplexa™ CongenitalCMV Direct Results | Detected | Not Detected | Total | ||
| Detected | 16 | 1 | 17 | 94.1%(16/17)95% CI:73% - 99% | 99.9%(1835/1836)95% CI:100% -100% |
| Not Detected | 1a | 1835 | 1836 | ||
| Total | 17 | 1836 | 1853 |
ª One (1) saliva swab specimen was negative by routine methodology.
PPA = Positive Percent Agreement, NPA = Negative Percent. 95% Cl = 95% Confidence Interval
The 95% confidence intervals (CI) were calculated following the Wilson Score method.
| Clinical Agreement | PPA | NPA | |||
|---|---|---|---|---|---|
| SimplexaTM CongenitalCMV Direct Results | Composite Reference Method (CRM) | ||||
| Detected | Not Detected | Total | |||
| Detected | 41 | 0 | 41 | 95.3%(41/43)95% CI:85% - 99% | 100%(1581/1581)95% CI:100% -100% |
| Not Detected | 2a | 1581 | 1583 | ||
| Total | 43 | 1581 | 1624 |
Table 4b. Simplexa™ Congenital CMV Direct Clinical Agreement - Urine (Prospective)
a Two (2) urine specimens were negative by routine methodology.
PPA = Positive Percent Agreement, NPA = Negative Percent Agreement. 95% Cl = 95% Confidence Interval
The 95% confidence intervals (CI) were calculated following the Wilson Score method.
REPRODUCIBILITY
Reproducibility for the Simplexa™ Congenital CMV Direct assay was evaluated. Three (3) investigative sites assessed the device's inter-site, inter-day and inter/intra-assay reproducibility. Each of the sites tested the Simplexa™ Congenital CMV Direct Positive Control, No Template Control (NTC), a negative urine sample, a negative saliva swab in UTM sample and eight (8) contrived samples spiked into a negative matrix of either saliva swabs in UTM or urine. The eight (8) contrived samples consisted of a low positive (LP) contrived at approximately 3X the limit of detection (LoD) and a medium positive (MP) contrived at approximately 10X LOD for each of the following: CMV strain AD169 in saliva swabs in UTM, CMV strain AD169 in urine, CMV strain Towne in saliva swabs in UTM, and CMV strain Towne in urine. Each contrived sample was prepared by spiking a specific concentration of the strain into CMV negative urine or a CMV neqative saliva swab in UTM. The samples were tested in quadruplicate on nine (9) different days. Each site had three (3) operators who each assayed the entire sample panel and Positive Control twice per day, for a total of two (2) sets of data per day on one (1) LIAISON® MDX instrument, per site. The combined results for all sites are presented in Table 5. The results show the reproducibility of the Simplexa™ Congenital CMV Direct % CV ranged between 0.4-1.6%.
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510(k) Summary
Simplexa™ Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022
Page 7 of 17
Table 5. Simplexa™ Congenital CMV Direct Reproducibility
| Site 1 | Site 2 | Site 3 | All Sites | |||||
|---|---|---|---|---|---|---|---|---|
| SampleCMV Strainand Matrix | Agreementwith expectedresults | Avg. Ct ±SD (%CV) | Agreementwith expectedresults | Avg. Ct ±SD (%CV) | Agreementwith expectedresults | Avg. Ct ±SD (%CV) | Agreementwithexpectedresults | Avg. Ct ±SD (%CV) |
| SalivaTowne_LP | 100.0%(36/36) | 34.3 ±0.32(0.9%) | 100.0%(36/36) | 34.5 ±0.26(0.7%) | 100.0%(36/36) | 34.3 ±0.32(0.9%) | 100.0%(108/108) | 34.4 ±0.32(0.9%) |
| SalivaTowne MP | 100.0%(36/36) | 32.2 ±0.25(0.8%) | 100.0%(36/36) | 32.5 ±0.21(0.6%) | 100.0%(36/36) | 32.2 ±0.22(0.7%) | 100.0%(108/108) | 32.3 ±0.25(0.8%) |
| SalivaAD-169_LP | 100.0%(36/36) | 33.4 ±0.34(1.0%) | 100.0%(36/36) | 33.8 ±0.23(0.7%) | 100.0%(36/36) | 33.5 ±0.25(0.8%) | 100.0%(108/108) | 33.6 ±0.32(1.0%) |
| SalivaAD-169 MP | 100.0%(36/36) | 32.2 ±0.22(0.7%) | 100.0%(36/36) | 32.3 ±0.27(0.8%) | 100.0%(36/36) | 32.1 ±0.15(0.5%) | 100.0%(108/108) | 32.2 ±0.23(0.7%) |
| UrineTowne_LP | 100.0%(36/36) | 33.9 ±0.51(1.5%) | 100.0%(36/36) | 34.0 ±0.48(1.4%) | 100.0%(36/36) | 33.9 ±0.45(1.3%) | 100.0%(108/108) | 34.0 ±0.48(1.4%) |
| UrineTowne MP | 100.0%(36/36) | 32.1 ±0.26(0.8%) | 100.0%(36/36) | 32.3 ±0.26(0.8%) | 100.0%(36/36) | 32.1 ±0.21(0.7%) | 100.0%(108/108) | 32.2 ±0.26(0.8%) |
| UrineAD-169 LP | 100.0%(36/36) | 35.9 ±0.57(1.6%) | 100.0%(36/36) | 36.1 ±0.59(1.6%) | 97.2%(35/36) | 35.9 ±0.46(1.3%) | 99.1%(107/108) | 36.0 ±0.54(1.5%) |
| UrineAD-169 MP | 100.0%(36/36) | 34.0 ±0.42(1.2%) | 100.0%(36/36) | 34.3 ±0.37(1.1%) | 100.0%(36/36) | 34.0 ±0.33(1.0%) | 100.0%(108/108) | 34.1 ±0.40(1.2%) |
| SalivaNegative* | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(108/108) | 0.0 ± 0.00(N/A%) |
| UrineNegative* | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(36/36) | 0.0 ±0.00(N/A%) | 100.0%(108/108) | 0.0 ±0.00(N/A%) |
| NTC (UTM) | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(36/36) | 0.0 ± 0.00(N/A%) | 100.0%(108/108) | 0.0 ±0.00(N/A%) |
| PC as is | 100.0%(36/36) | 29.7 ±0.14(0.5%) | 100.0%(36/36) | 29.9 ±0.13(0.4%) | 100.0%(36/36) | 29.6 ±0.22(0.7%) | 100.0%(108/108) | 29.7 ±0.20(0.7%) |
LP = Low Positive, MP = Moderate Positive, UTM = Universal Transport Media, NTC = No Template Control, SD = Standard Deviation, %CV = Percent Coefficient of Variation, Ct = Cycle Threshold
*Expected result for these samples is negative.
ANALYTICAL SENSITIVITY/LIMIT OF DETECTION
The limit of detection (LoD) was determined for the Simplexa™ Congenital CMV Direct assay using quantified stocks of three (3) CMV strains (AD169, Towne and Merlin) serially diluted in negative human saliva swab and urine matrices. The LoD was determined to be the lowest concentration that could be detected positive > 95% of the time. The LoD for each matrix is presented in Tables 6a and 6b.
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| CMV strain | LoD - Saliva in 1mL UTM | LoD - Saliva in 3mL UTM | LoD - Saliva in 3mL M4RT | |||
|---|---|---|---|---|---|---|
| Copies/mLin Saliva | Copies/mLin UTM | Copies/mLin saliva | Copies/mLin UTM | Copies/mLin saliva | Copies/mLin M4RT | |
| AD-169 | 6,750 | 500 | 19,250 | 500 | 19,250 | 500 |
| Towne | 6,750 | 500 | 19,250 | 500 | 19,250 | 500 |
| CMV strain | IU/mL insaliva | IU/mL inUTM | IU/mL insaliva | IU/mL inUTM | IU/mL insaliva | IU/mL inM4RT |
| Merlin | 6,750 | 500 | 19,250 | 500 | 19,250 | 500 |
| Table 6b. Simplexa™ Congenital CMV Direct Limit of Detection - Urine | ||
|---|---|---|
| CMV strain | Copies /mL |
|---|---|
| AD-169 | 400 Copies/mL |
| Towne | 800 Copies/mL |
| CMV strain | IU/mL |
| Merlin | 6,400 IU/mL |
ANALYTICAL REACTIVITY/CROSS REACTIVITY
Analytical Reactivity
The analytical reactivity of the Simplexa™ Congenital CMV Direct assay was evaluated using different strains/genotypes of CMV that were not used in the determination of the limit of detection (LoD) for the assay. Quantified CMV was spiked at 1x LoD into negative adult saliva swab and negative urine from neonates less than 21 days of age. For the saliva swabs the preparations were spiked onto a flocked swab and transferred to each tube of UTM. The results are presented in Table 7. No genotype 4 (gB4) strains were available for testing. In addition to the strains that were physically tested, in silico BLAST analysis demonstrated that the assay should detect at least 327 CMV sequences available in the NCBI database, including the four (4) CMV genotypes gB1, gB2, gB3 and gB4.
Table 7. Simplexa™ Congenital CMV Direct Analytical Reactivity
| CMV (gB3) Strain andMatrix | Agreement withExpected Results(#Detected/#Total) |
|---|---|
| CMV Toledo Strain(Saliva Swab) | 100% (10/10) |
| CMV Toledo Strain(Urine) | 100% (10/10) |
Cross-Reactivity (Analytical Specificity)
The Simplexa™ Congenital CMV Direct assay's analytical specificity was evaluated by testing the ability of the assay to exclusively identify CMV virus with no cross-reactivity to organisms that are closely related, cause similar clinical symptoms or may be present in saliva swabs and urine. Forty-one (41) potential crossreactants were spiked into negative saliva swab and 13 potential cross reactants were spiked into negative
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Simplexa™ Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022 Page 9 of 17
urine. The samples were assayed in triplicate. No cross-reactivity was observed. The results are presented in Tables 8a and 8b.
| No. | Organism | Tested Concentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|
| 1 | Acinetobacter baumannii | 1x106 CFU/mL | 100.0% (3/3) |
| 2 | Actinomyces odontolyticus | 1x106 CFU/mL | 100.0% (3/3) |
| 3 | Bordetella pertussis | 1x106 CFU/mL | 100.0% (3/3) |
| 4 | Coronavirus 229E | 1x105 TCID50/mL | 100.0% (3/3) |
| 5 | CoxsackievirusA9 | 1x105 TCID50/mL | 100.0% (3/3) |
| 6 | Epstein-Barr Virus | 1x105 IU/mL | 100.0% (3/3) |
| 7 | Enterovirus 71* | 1x104 TCID50/mL | 100.0% (3/3) |
| 8 | FLU A/ Michigan/45/2015 | 1x105 Cps/mL | 100.0% (3/3) |
| 9 | FLU B/ Phuket/3073/2013 | 1x105 Cps/mL | 100.0% (3/3) |
| 10 | Fusobacterium nucleatum | 1x106 CFU/mL | 100.0% (3/3) |
| 11 | Adenovirus (C1) | 1x105 Cps/mL | 100.0% (3/3) |
| 12 | Haemophilus influenza | 1x106 CFU/mL | 100.0% (3/3) |
| 13 | Haemophilus parainfluenzae | 1x106 CFU/mL | 100.0% (3/3) |
| 14 | Herpes Simplex Virus 1 | 1x105 IU/mL | 100.0% (3/3) |
| 15 | Human herpesvirus 6A | 1x105 Cps/mL | 100.0% (3/3) |
| 16 | Human herpesvirus 6B | 1x105 Cps/mL | 100.0% (3/3) |
| 17 | Human herpesvirus 7 | 1x105 Cps/mL | 100.0% (3/3) |
| 18 | Human herpesvirus 8 | 1x105 Cps/mL | 100.0% (3/3) |
| 19 | Human Genomic DNA | 1x106 Cps/mL | 100.0% (3/3) |
| 20 | Human metapneumovirus | 1x105 TCID50/mL | 100.0% (3/3) |
| 21 | Klebsiella oxytoca | 1x106 CFU/mL | 100.0% (3/3) |
| 22 | Klebsiella pneumoniae | 1x106 CFU/mL | 100.0% (3/3) |
| 23 | Moraxella catarrhalis | 1x106 CFU/mL | 100.0% (3/3) |
| 24 | Mycoplasma pneumoniae | 1x106 CCU/mL | 100.0% (3/3) |
| 25 | Parainfluenza virus 1 | 1x105 U/mL | 100.0% (3/3) |
| 26 | Parainfluenza virus 2 | 1x105 U/mL | 100.0% (3/3) |
| 27 | Parainfluenza virus 3 | 1x105 U/mL | 100.0% (3/3) |
| 28 | Porphyromonas gingivalis | 1x106 CFU/mL | 100.0% (3/3) |
| 29 | Pseudomonas aeruginosa | 1x106 CFU/mL | 100.0% (3/3) |
| 30 | Respiratory syncytial virus A | 1x105 IU/mL | 100.0% (3/3) |
| 31 | Respiratory syncytial virus B | 1x105 TCID50/mL | 100.0% (3/3) |
| 32 | Rhinovirus | 1x105 U/mL | 100.0% (3/3) |
| 33 | Staphylococcus aureus | 1x106 CFU/mL | 100.0% (3/3) |
| 34 | Staphylococcus epidermidis | 1x106 CFU/mL | 100.0% (3/3) |
| 35 | Streptococcus anginosus | 1x106 CFU/mL | 100.0% (3/3) |
| 36 | Streptococcus oralis | 1x106 CFU/mL | 100.0% (3/3) |
| No. | Organism | Tested Concentration | % Agreement**(# Expected Results/ #Tested) |
| 37 | Streptococcus mitis | 1x106 CFU/mL | 100.0% (3/3) |
| 38 | Streptococcus pneumoniae | 1x106 CFU/mL | 100.0% (3/3) |
| 39 | Streptococcus salivarius | 1x106 CFU/mL | 100.0% (3/3) |
| 40 | Streptococcus sanguinis | 1x106 CFU/mL | 100.0% (3/3) |
| 41 | Varicella Zoster Virus | 1x105 Cps/mL | 100.0% (3/3) |
Table 8a. Simplexa™ Congenital CMV Direct Cross-Reactivity (Analytical Specificity) - Saliva Swab
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** Expected result for all organisms is negative.
*Enterovirus 71 was tested at a concentration lower than 1x10 TCIDs/mL due to the low concentration of the virus stock that was available. In silico (BLAST) analysis was also performed. The BLAST analysis showed that no crossreactivity is expected with this microorganism.
| No. | Organism | Tested Concentration | % Agreement(# Expected Results/ #Tested) |
|---|---|---|---|
| 1 | Candida albicans | 1x106 CFU/mL | 100.0% (3/3) |
| 2 | Enterobacter aerogenes | 1x106 CFU/mL | 100.0% (3/3) |
| 3 | Enterobacter cloacae | 1x106 CFU/mL | 100.0% (3/3) |
| 4 | Enterococcus faecium | 1x106 CFU/mL | 100.0% (3/3) |
| 5 | Enterococcus faecalis | 1x106 CFU/mL | 100.0% (3/3) |
| 6 | Escherichia coli | 1x106 CFU/mL | 100.0% (3/3) |
| 7 | Herpes Simplex Virus 2 | 1x105 cps/mL | 100.0% (3/3) |
| 8 | Lactobacillus acidophilus | 1x106 CFU/mL | 100.0% (3/3) |
| 9 | Morganella morganii | 1x106 CFU/mL | 100.0% (3/3) |
| 10 | Proteus mirabilis | 1x106 CFU/mL | 100.0% (3/3) |
| 11 | Proteus vulgaris | 1x106 CFU/mL | 100.0% (3/3) |
| 12 | Streptococcus agalactiae (GBS) | 1x106 CFU/mL | 100.0% (3/3) |
| 13 | Enterovirus 71* | 1x104 TCID50/mL | 100.0% (3/3) |
Table 8b. Simplexa™ Congenital CMV Direct Cross-Reactivity (Analytical Specificity) - Urine
*Expected result for all organisms is negative.
*Enterovirus 71 was tested at a concentration lower than 1x10 TCIDs/mL due to the virus stock that was available. In silico (BLAST) analysis was also performed. The BLAST analysis showed that no cross-reactivity is expected with this microorganism.
INTERFERENCE
The performance of the Simplexa™ Congenital CMV Direct assay was evaluated with potentially interfering substances that may be present in saliva swabs and urine samples at the concentrations indicated in the table below. A total of 17 potentially interfering substances were tested for saliva swabs and seven (7) potentially interfering substances were tested for urine in a low positive CMV sample at approximately three times the limit of detection (3X LoD) in saliva swab and urine matrices and assayed in triplicate. No interference was observed. The results are presented in Tables 9a and 9b.
Table 9a. Simplexa™ Congenital CMV Direct Interference for saliva swab
| No. | Potential Interferent | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|---|
| 1 | Acetylsalicylic acid | AD169 | 0.65 mg/mL | 100.0% (3/3) |
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| No. | Potential Interferent | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|---|
| 1 | Towne | 0.65 mg/mL | 100.0% (3/3) | |
| 2 | Breast milk | AD169 | 10% (v/v) | 100.0% (3/3) |
| Towne | 10% (v/v) | 100.0% (3/3) | ||
| 3 | Caffeine | AD169 | 0.06 mg/mL | 100.0% (3/3) |
| Towne | 0.06 mg/mL | 100.0% (3/3) | ||
| 4 | Casein | AD169 | 10 mg/mL | 100.0% (3/3) |
| Towne | 10 mg/mL | 100.0% (3/3) | ||
| 5 | Enfamil Poly-vi-sol withIron | AD169 | 1.5 mg/mL | 100.0% (3/3) |
| Towne | 1.5 mg/mL | 100.0% (3/3) | ||
| 6 | Enfamil™ formula neuropro® | AD169 | 10% (v/v) | 100.0% (3/3) |
| Towne | 10% (v/v) | 100.0% (3/3) | ||
| 7 | Enfamil™ Tri-Vi-Sol® | AD169 | 8% (v/v) | 100.0% (3/3) |
| Towne | 8% (v/v) | 100.0% (3/3) | ||
| 8 | Gaviscon® (SodiumAlginate) | AD169 | 1.2 mg/mL | 100.0% (3/3) |
| Towne | 1.2 mg/mL | 100.0% (3/3) | ||
| 9 | Infants' Pain & Fever(Acetaminophen) | AD169 | 0.4 mg/mL | 100.0% (3/3) |
| Towne | 0.2 mg/mL | 100.0% (3/3) | ||
| 10 | Infants' Mylicon® GasRelief (Simethicone) | AD169 | 6.7 mg/mL | 100.0% (3/3) |
| Towne | 6.7 mg/mL | 100.0% (3/3) | ||
| 11 | Little Remedies SalineDrops | AD169 | 10% (v/v) | 100.0% (3/3) |
| Towne | 10% (v/v) | 100.0% (3/3) | ||
| 12 | Motrin Infant Drops(Infants' Ibuprofen) | AD169 | 0.5 mg/mL | 100.0% (3/3) |
| Towne | 0.5 mg/mL | 100.0% (3/3) | ||
| 13 | Mucin | AD169 | 25 mg/mL | 100.0% (3/3) |
| Towne | 25 mg/mL | 100.0% (3/3) | ||
| 14 | Nystatin | AD169 | 1,727 U/mL | 100.0% (3/3) |
| Towne | 1,727 U/mL | 100.0% (3/3) | ||
| 15 | Prednisone | AD169 | 0.0003 mg/mL | 100.0% (3/3) |
| Towne | 0.0003 mg/mL | 100.0% (3/3) | ||
| 16 | White Blood Cell | AD169 | 10% (v/v) | 100.0% (3/3) |
| Towne | 10% (v/v) | 100.0% (3/3) | ||
| 17 | Whole blood | AD169 | 10% (v/v) | 100.0% (3/3) |
| Towne | 10% (v/v) | 100.0% (3/3) |
**Expected result for all potential interferents is positive.
| Table 9b. Simplexa™ Congenital CMV Direct Interference for urine | |||||
|---|---|---|---|---|---|
| -- | -- | -- | -- | ------------------------------------------------------------------- | -- |
| No. | Organism | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|---|
| 1 | Baby powder | AD169 | 10% (w/v) | 100% (3/3) |
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Simplexa™ Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2. 2022
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| No. | Organism | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|---|
| Towne | 10% (w/v) | 100% (3/3) | ||
| 2 | Johnson's Baby Oil(Mineral Oil) | AD169 | 10% (v/v) | 100% (3/3) |
| Towne | 10% (v/v) | 100% (3/3) | ||
| Meconium | AD169 | 0.5% (w/v) | 100% (3/3) | |
| 3 | Towne | 1% (w/v) | 100% (3/3) | |
| Moist Towelettes withBenzalkonium Chloride | AD169 | 10% (w/v) | 100% (3/3) | |
| 4 | Towne | 10% (w/v) | 100% (3/3) | |
| 5 | Nystatin | AD169 | 0.3mg/mL | 100% (3/3) |
| Towne | 0.3mg/mL | 100% (3/3) | ||
| 6 | Stool | AD169 | 2% (w/v) | 100% (3/3) |
| Towne | 2% (w/v) | 100% (3/3) | ||
| 7 | Whole blood | AD169 | 5% (v/v) | 100% (3/3) |
| Towne | 10% (v/v) | 100% (3/3) |
**Expected result for all potential interferents is positive.
INHIBITION BY OTHER MICROORGANISMS
The Simplexa™ Congenital CMV Direct assay was evaluated by testing the ability to identify CMV virus when other potentially inhibitory organisms are present. A panel of forty-one (41) potentially inhibitory organisms was individually spiked into a pool with a low concentration of CMV at approximately three times the limit of detection (3X LoD) in saliva. Thirteen (13) potentially inhibitory organisms were individually spiked into a pool with a low concentration of CMV at approximately three the limit of detection (3X LoD) in urine. Potentially inhibiting organisms were tested at the concentrations specified in Tables 10a and 10b. No inhibition by other organisms was observed.
Table 10a Simplexa™ Congenital CMV Direct Microbial Interference – Saliva Swab
| No. | Organism | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|---|
| 1 | Acinetobacter baumannii | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 2 | Actinomyces odontolyticus | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 3 | Bordetella pertussis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 4 | Coronavirus 229E | AD169 | 1x105 TCID50/mL | 100.0% (3/3) |
| Towne | 1x105 TCID50/mL | 100.0% (3/3) | ||
| 5 | CoxsackievirusA9 | AD169 | 1x105 TCID50/mL | 100.0% (3/3) |
| Towne | 1x105 TCID50/mL | 100.0% (3/3) | ||
| 6 | Epstein-Barr Virus | AD169 | 1x105 IU/mL | 100.0% (3/3) |
| Towne | 1x105 IU/mL | 100.0% (3/3) | ||
| 7 | Enterovirus 71* | AD169 | 1x104 TCID50/mL | 100.0% (3/3) |
| No. | Organism | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
| 8 | FLU A/ Michigan/45/2015 | Towne | 1x104 TCID50/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 9 | FLU B/ Phuket/3073/2013 | Towne | 1x105 Cps/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 10 | Fusobacterium nucleatum | Towne | 1x106 CFU/mL | 100.0% (3/3) |
| AD169 | 1x106 CFU/mL | 100.0% (3/3) | ||
| 11 | Adenovirus (C1) | Towne | 1x105 Cps/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 12 | Haemophilus influenza | Towne | 1x106 CFU/mL | 100.0% (3/3) |
| AD169 | 1x106 CFU/mL | 100.0% (3/3) | ||
| 13 | Haemophilus parainfluen zae | Towne | 1x106 CFU/mL | 100.0% (3/3) |
| AD169 | 1x106 CFU/mL | 100.0% (3/3) | ||
| 14 | Herpes Simplex Virus 1 | Towne | 1x105 IU/mL | 100.0% (3/3) |
| AD169 | 1x105 IU/mL | 100.0% (3/3) | ||
| 15 | Human herpesvirus 6A | Towne | 1x105 Cps/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 16 | Human herpesvirus 6B | Towne | 1x105 Cps/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 17 | Human herpesvirus 7 | Towne | 1x105 Cps/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 18 | Human herpesvirus 8 | Towne | 1x105 Cps/mL | 100.0% (3/3) |
| AD169 | 1x105 Cps/mL | 100.0% (3/3) | ||
| 19 | Human Genomic DNA | Towne | 1x106 Cps/mL | 100.0% (3/3) |
| AD169 | 1x106 Cps/mL | 100.0% (3/3) | ||
| 20 | Human metapneumovirus | Towne | 1x105 TCID50/mL | 100.0% (3/3) |
| AD169 | 1x105 TCID50/mL | 100.0% (3/3) | ||
| 21 | Klebsiella oxytoca | Towne | 1x106 CFU/mL | 100.0% (3/3) |
| AD169 | 1x106 CFU/mL | 100.0% (3/3) | ||
| 22 | Klebsiella pneumoniae | Towne | 1x106 CFU/mL | 100.0% (3/3) |
| AD169 | 1x106 CFU/mL | 100.0% (3/3) | ||
| 23 | Moraxella catarrhalis | Towne | 1x106 CFU/mL | 100.0% (3/3) |
| AD169 | 1x106 CFU/mL | 100.0% (3/3) | ||
| 24 | Mycoplasma pneumoniae | Towne | 1x106 CCU/mL | 100.0% (3/3) |
| AD169 | 1x106 CCU/mL | 100.0% (3/3) | ||
| 25 | Parainfluenza virus 1 | Towne | 1x105 U/mL | 100.0% (3/3) |
| AD169 | 1x105 U/mL | 100.0% (3/3) | ||
| 26 | Parainfluenza virus 2 | Towne | 1x105 U/mL | 100.0% (3/3) |
| AD169 | 1x105 U/mL | 100.0% (3/3) | ||
| 27 | Parainfluenza virus 3 | Towne | 1x105 U/mL | 100.0% (3/3) |
| AD169 | 1x105 U/mL | 100.0% (3/3) | ||
| No. | Organism | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
| 28 | Porphyromonas gingivalis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 29 | Pseudomonas aeruginosa | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 30 | Respiratory syncytial virus A | AD169 | 1x105 IU/mL | 100.0% (3/3) |
| Towne | 1x105 IU/mL | 100.0% (3/3) | ||
| 31 | Respiratory syncytial virus B | AD169 | 1x105 TCID50/mL | 100.0% (3/3) |
| Towne | 1x105 TCID50/mL | 100.0% (3/3) | ||
| 32 | Rhinovirus | AD169 | 1x105 U/mL | 100.0% (3/3) |
| Towne | 1x105 U/mL | 100.0% (3/3) | ||
| 33 | Staphylococcus aureus | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 34 | Staphylococcus epidermidis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 35 | Streptococcus anginosus | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 36 | Streptococcus oralis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 37 | Streptococcus mitis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 38 | Streptococcus pneumoniae | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 39 | Streptococcus salivarius | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 40 | Streptococcus sanguinis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 41 | Varicella Zoster Virus | AD169 | 1x105 Cps/mL | 100.0% (3/3) |
| Towne | 1x105 Cps/mL | 100.0% (3/3) |
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510(k) Simplexa™ Congenital CMV Direct MOL2250
Simplexa™ Congenital CMV Direct MOL2250
Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022
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Image /page/16/Picture/1 description: The image contains the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color.
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*Expected result for all organisms is positive.
- Enterovirus 71 was tested at a concentration lower than the 1x105 TCID50/mL due to the virus stock that was available. In silico (BLAST)
analysis was also performed. The results of the BLAST analysis showed that no microbial inhibition is expected with this microorganism.
| ----------------------------------- | AMI OL----- | Tested | % Agreeme14 F ------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
|---|
Table 10b Simplexa™ Congenital CMV Direct Microbial Interference - Urine
| No. | Organism | CMV Strain | TestedConcentration | % Agreement(# Expected Results/ #Tested) |
|---|---|---|---|---|
| 1 | Candida albicans | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) |
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Simplexa™ Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. M 2260 Nov 2. 2022
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| No. | Organism | CMV Strain | TestedConcentration | % Agreement**(# Expected Results/ #Tested) |
|---|---|---|---|---|
| 2 | Enterobacter aerogenes | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 3 | Enterobacter cloacae | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 4 | Enterococcus faecium | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 5 | Enterococcus faecalis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 6 | Escherichia coli ATCC | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 7 | Herpes Simplex Virus 2 | AD169 | 1x105 Cps/mL | 100.0% (3/3) |
| Towne | 1x105 Cps/mL | 100.0% (3/3) | ||
| 8 | Lactobacillus acidophilus | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 9 | Morganella morganii | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 10 | Proteus mirabilis | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 11 | Proteus vulgaris | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 12 | Streptococcusagalactiae (GBS) | AD169 | 1x106 CFU/mL | 100.0% (3/3) |
| Towne | 1x106 CFU/mL | 100.0% (3/3) | ||
| 13 | Enterovirus 71* | AD169 | 1x104 TCID50/mL | 100.0% (3/3) |
| Towne | 1x104 TCID50/mL | 100.0% (3/3) |
** Expected result for all organisms is positive.
- Enterovirus 71 was tested at a concentration lower than the 1x10s TCIDs; mL due to the virus stock that was available. In silico (BLAST) analysis was also performed. The BLAST analysis showed that no microbial inhibition is expected with this microorganism.
CARRY-OVER CONTAMINATION
Amplification carry-over for the Simplexa™ assays has been assessed. The study was designed by alternately placing high positive and negative samples on each disc. No evidence of carry-over contamination was observed.
The performance of the Simplexa™ Congenital CMV Direct assay was established in a clinical study that included two (2) cohorts based on sample status. Specifically, prospective samples from infants less than twenty-one (21) days of age were tested in the clinical agreement study.
EXPECTED VALUES
The prevalence of CMV as determined by the Simplexa™ Congenital CMV Direct assay in a multi-site clinical study with prospectively collected specimens was 1.19% for saliva swabs and 2.59% for urine. Table
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Simplexa™ Congenital CMV Direct MOL2250 Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022 Page 16 of 17
8 shows the prevalence of saliva swabs by collection site and Table 9 shows the prevalence of urine by collection site.
Table 8. Prospective Results: Simplexa™ Congenital CMV Direct Expected Values for Saliva Swabs by Collection Site
| Site ID | Total Specimens | Simplexa™ CongenitalCMV Direct Detected | CMV Prevalence |
|---|---|---|---|
| 7 | 142 | 0 | 0.00% |
| 8 | 16 | 1 | 6.25% |
| 9 | 632 | 3 | 0.47% |
| 10 | 9 | 0 | 0.00% |
| 12 | 14 | 0 | 0.00% |
| 13 | 11 | 1 | 9.09% |
| 14 | 9 | 0 | 0.00% |
| 15 | 10 | 0 | 0.00% |
| 18 | 8 | 0 | 0.00% |
| 19 | 1002 | 12 | 1.20% |
| All | 1853 | 17 | 0.92% |
Table 9. Prospective Results: Simplexa™ Congenital CMV Direct Expected Values for Urine by Collection Site
| Site ID | All | Simplexa™ CongenitalCMV Direct Detected | CMV Prevalence |
|---|---|---|---|
| 5 | 171 | 2 | 1.17% |
| 6 | 1336 | 36 | 2.69% |
| 7 | 5 | 0 | 0.00% |
| 8 | 47 | 2 | 4.26% |
| 10 | 8 | 0 | 0.00% |
| 12 | 13 | 0 | 0.00% |
| 13 | 11 | 1 | 9.09% |
| 14 | 9 | 0 | 0.00% |
| 15 | 10 | 0 | 0.00% |
| 18 | 11 | 0 | 0.00% |
| 19 | 3 | 0 | 0.00% |
| All | 1624 | 41 | 2.52% |
Conclusion
The analytical and method comparison studies have demonstrated that the Simplexa™ Congenital CMV Direct is Substantially Equivalent to the predicate device (DEN180040).The device labeling is compliant with 21 CFR § 809.10.
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Simplexa™ Congenital CMV Positive Control Pack. MOL2260 Nov 2, 2022 Page 17 of 17
§ 866.3181 Cytomegalovirus nucleic acid detection device for congenital cytomegalovirus infection.
(a)
Identification. A cytomegalovirus nucleic acid detection device for congenital cytomegalovirus infection is an in vitro diagnostic device intended for the qualitative detection of cytomegalovirus DNA in clinical samples from newborn babies to aid in the diagnosis of congenital cytomegalovirus infection. Negative results do not preclude infection and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results should be interpreted with consideration of other clinical information and laboratory findings and should not be used as the sole basis for treatment or other patient management decisions.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The labeling required under § 809.10(b) of this chapter must include:
(i) An intended use with a detailed description of what the device detects, the type of results provided to the user, the clinical indications appropriate for test use, and the specific population(s) to be tested.
(ii) A detailed device description, including all device components, instrument requirements, ancillary reagents required but not provided, and an explanation of the methodology, including all pre-analytical methods for specimen processing.
(iii) Performance characteristics from analytical and clinical studies required under paragraphs (b)(2)(ii) and (iii) of this section.
(iv) A detailed explanation of the interpretation of results and criteria for validity of results.
(v) A limiting statement that device results are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions.
(vi) As applicable, a limiting statement and specific sample collection recommendations to indicate that breast milk can result in false positive results for saliva samples if samples are collected less than 1 hour after breastfeeding. Sample collection a minimum of 1 hour from breastfeeding must be recommended.
(vii) Detailed instructions for use that minimize the risk of generating a false result.
(2) Design verification and validation must include:
(i) Detailed device description documentation, including methodology from obtaining sample to result, design of primer/probe sequences, rationale for sequence selection, and computational path from collected raw data to reported result (
e.g., how collected raw signals are converted into a reported result).(ii) Detailed documentation of analytical studies including characterization of the cutoff, analytical sensitivity (limit of detection), inclusivity, reproducibility, interference, cross reactivity, instrument and method carryover/cross contamination, and sample stability and handling.
(iii) Detailed documentation from a clinical study documenting sensitivity and specificity of the device; if the number of positive samples in the clinical study is insufficient to properly estimate device sensitivity, additional pre-selected positive samples must be evaluated to supplement the study. Clinical study subjects must be consistent with the intended use population (
i.e., infants younger than 21 days of age), and device results must be compared to FDA-accepted comparator methods. Documentation from the clinical study must include the clinical study protocol, the clinical study report, testing results, and results of all statistical analyses.(iv) Detailed documentation for device software, including software applications and hardware-based devices that incorporate software.