(344 days)
iClean Viral Transport System (VTM-RT) is intended for the collection and transport of clinical specimens containing respiratory viruses, Chlamydiae, or Mycoplasma hominis from the collection site to the testing laboratory. The collection system is a culture based media that is intended to be used with standard laboratory examination, culture or with other assays that utilize stable recoverable infectious viral particles or bacteria.
iClean Viral Transport System (VTM-RT) is intended for the collection and transport of clinical specimens containing respiratory viruses, Chlamydiae, or Mycoplasma hominis from the collection site to the testing laboratory. iClean Viral Transport System (VTM-RT) includes a plastic screw-cap tube with conical bottom containing 3mL transport medium. iClean Viral Transport System (VTM-RT) tubes can be supplied alone, or in a kit format with a flocked swab in a sterile peel pouch (or 100 kits packed together in a color box). iClean Viral Transport System (VTM-RT) medium is stable at room temperature and consists of: Hanks balanced salt solution (HBSS), Bovine Serum Albumin (BSA), Gentamicin sulfate, Amphotericin B, Colistin, L-glutamic acid, HEPES buffer and Phenol red.
Here's an analysis of the provided text, focusing on the acceptance criteria and the study demonstrating that the iClean Viral Transport System (VTM-RT kit) meets those criteria.
Please note that the document describes a transport medium, not an AI-powered device. Therefore, several sections of your request (e.g., sample size for the test set, number of experts, adjudication method, MRMC comparative effectiveness study, standalone performance, sample size for the training set, how ground truth for training set was established) are not applicable as they relate specifically to AI/ML device studies.
Acceptance Criteria and Device Performance for iClean Viral Transport System (VTM-RT kit)
The iClean VTM-RT kit is a transport medium, and its performance is assessed by its ability to safely preserve and transport various microorganisms while maintaining their viability. The acceptance criteria and reported performance are based on recovery studies for viral and bacterial test strains, and stability studies examining physical and chemical properties of the medium over time.
1. Table of Acceptance Criteria and Reported Device Performance
Acceptance Criteria for iClean Viral Transport System (VTM-RT kit)
| Category | Specific Criterion | Reported Device Performance |
|---|---|---|
| Shelf-Life Stability | Appearance: Clear (no turbidity, cloudiness, precipitation), pink color. | All results were acceptable; visually inspected media at T=0, 1, 3, 6, 9, 12 months maintained clear, pink appearance. |
| Volume: 3.0 mL per tube. | All results acceptable; media volume assessed at T=0, 1, 3, 6, 9, 12 months ensured each tube was filled to 3.0mL. | |
| pH: 7.4 ± 0.4. | All results acceptable; pH measured at T=0, 1, 3, 6, 9, 12 months across three lots remained within 7.4 ± 0.4. | |
| Antibiotic Stability: Inhibit bacterial/yeast growth (no growth at 24/48 hrs after inoculation). | Inoculated media with S. aureus, E. coli, C. albicans showed growth at T=0, but no growth at 24 and 48 hours for all plates, demonstrating effective antibiotic stability over time. | |
| Sterility: Absence of bacteria and fungi after filtration and aseptic filling. | Quality control process confirmed absence of growth of bacteria and fungi on nutrient agar and Sabouraud media plates after incubation, validating aseptic status. | |
| Microorganism Recovery | Viral Recovery: Acceptable preservation of Fluorescent Foci Count for Influenza A, Parainfluenza 3, Respiratory Syncytial Virus, Chlamydia pneumoniae, and C. trachomatis over 48 hours at 4°C and 25°C. | At 4°C: Declines in foci count range from -6% to 92% over 48 hours, with most below 60%. The document does not specify a numerical acceptance threshold for percent decline, but implies that these levels demonstrate "recovery." At 25°C: Declines in foci count range from -194% (likely an error or high variability) to 96% over 48 hours. Most declines are significantly higher than at 4°C, indicating sensitivity to higher temperatures, but still demonstrating recovery. The wide range and some negative declines suggest variability or specific strain behavior. |
| Bacterial Recovery (Roll-Plate): ≥ 10 CFU recovery after specified maintenance time. | All reported average CFU/roll-plate values for Mycoplasma hominis at 24 and 48 hours (both 2-8°C and 22-25°C) are well above 10 CFU, ranging from 72 to 289 CFU at 48 hours, thus meeting the criteria. | |
| Bacterial Recovery (Swab Elution): No more than a 3 log10 change in CFU count between 0-time and 48 hours. | For Mycoplasma hominis, the change in log10 CFU from 0 to 48 hours ranges from -0.5 to -1.0 at 2-8°C, and from -0.7 to -1.3 (or showing gains/inconsistent values for some logs that are likely typos, e.g., 2.1x10^6 and 1.1x10^6 in old lot) at 22-25°C. These values are all well within the acceptable limit of "no more than a 3 log10 change", with reported changes typically around 1 log10 or less. |
Study Proving Device Meets Acceptance Criteria
The study conducted is a series of in vitro performance and stability tests designed to assess the iClean VTM-RT kit's ability to maintain the viability of specified microorganisms and the integrity of the transport medium over its claimed shelf-life.
The general conclusion, as stated in the document, is: "The iClean VTM demonstrated the recovery of tested viruses, Chlamydia pneumoniae, C. trachomatis and Mycoplasma hominis in all replicates at tested incubation times and storage conditions." This statement implies that all acceptance criteria were met, based on their internal definitions of "recovery" and "stability."
2. Sample Size Used for the Test Set and Data Provenance
- Sample Size (Test Set):
- Shelf-Life Studies: Three lots of iClean VTM-RT media were used for appearance, volume, pH, and antibiotic stability testing.
- Recovery Studies: Nine lots of media were used, categorized as "New" (newly manufactured), "Mid" (middle-aged, ~5 months old), and "Old" (about to expire or recently expired). The tables show 3 lots for each age category for a total of 9 lots. For each lot and condition (time point, temperature), multiple replicates were used (e.g., swabs in triplicate for bacterial studies, 15 tubes for pH).
- Data Provenance: The document does not explicitly state the country of origin for the data collection. However, the manufacturer is Huachenyang (Shenzhen) Technology Co. Ltd. (China), suggesting the studies were likely conducted in China or a related facility. The studies are retrospective in the sense that they are laboratory evaluations of manufactured product lots, not prospective patient studies.
3. Number of Experts and Qualifications (Not Applicable)
This information is relevant for AI/ML diagnostic devices requiring expert interpretation. For a transport medium, the assessment is based on quantifiable laboratory measures of microorganism viability and chemical/physical stability, not expert interpretation of results.
4. Adjudication Method (Not Applicable)
This is relevant for AI/ML diagnostic devices where multiple experts might disagree on ground truth. For a transport medium, the "ground truth" is based on objective laboratory measurements.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study (Not Applicable)
This is specific to AI/ML devices that assist human readers. The iClean VTM-RT is a laboratory instrument/reagent, not an AI system.
6. Standalone Performance Study (i.e., algorithm only without human-in-the-loop performance) (Not Applicable)
This is specific to AI/ML devices. The iClean VTM-RT is a transport medium, and its performance is inherently "standalone" in the sense that it functions based on its chemical and physical properties to preserve microorganisms.
7. Type of Ground Truth Used
The ground truth used for these studies is based on:
- Viral Recovery: Fluorescent Foci Count method. This is a quantitative measure of infectious viral particles after storage.
- Bacterial Recovery (Roll-Plate): Colony-Forming Units (CFU) counts directly from plates rolled with the swab.
- Bacterial Recovery (Swab Elution): CFU counts from serial dilutions of the eluate.
- Stability Studies: Objective measurements of pH, visual inspection for appearance, volume measurements, and microbial growth/no-growth for antibiotic efficacy.
These are all laboratory-based, quantitative measures of the viability of the target microorganisms and the integrity of the medium.
8. Sample Size for the Training Set (Not Applicable)
This is relevant for AI/ML models that require a training dataset. The iClean VTM-RT is a chemical/biological product, not an AI model.
9. How the Ground Truth for the Training Set Was Established (Not Applicable)
As there is no AI/ML training set, this question is not applicable.
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August 18, 2022
Huachenyang (Shenzhen) Technology Co. Ltd. % Prithul Bom Most Responsible Person Regulatory Technology Services, LLC 1000 Westgate Drive, Suite 510k Saint Paul, Minnesota 55114
Re: K212856
Trade/Device Name: iClean Viral Transport System (VTM-RT kit) Regulation Number: 21 CFR 866.2390 Regulation Name: Transport Culture Medium Regulatory Class: Class I, reserved Product Code: JSM Dated: September 7, 2021 Received: September 8, 2021
Dear Prithul Bom:
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.
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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 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 medical devices and radiation-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.
for Uwe Scherf, M.Sc., Ph.D. 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) K212856
Device Name iClean Viral Transport System (VTM-RT kit)
Indications for Use (Describe)
iClean Viral Transport System (VTM-RT) is intended for the collection and transport of clinical specimens containing respiratory viruses, Chlamydiae, or Mycoplasma hominis from the to the testing laboratory. The collection system is a culture based media that is intended to be used with standard laboratory examination, culture or with other assays that utilize stable recoverable infectious viral particles or bacteria.
| 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) |
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510(k) Summary - K212856
In accordance with the Food and Drug Administration Rule to implement provisions of the Safe Medical Devices Act of 1990 and in conformance with 21 CFR 807.92, this information serves as a Summary of Safety and Effectiveness for the iClean Viral Transport System (VTM- RT).
| Submitted by | Huachenyang (Shenzen) Technology Co., Ltd. |
|---|---|
| Contact Person | Gong Zanbin, Production & Regulation Manager |
| Huachenyang (Shenzen) Technology Co., Ltd.F8/Bldg. 4, Hengchangrong High Industrial Park, No.128 EastRoad, Shangnan, Shajing Street, Baoan, Shenzhen, China Email:info@chenyanglobal.com; c06@chenyanglobal.com TelephoneNumber: + 86-755-27393226Fax Number: + 86-755-27381080 | |
| Proprietary Name | iClean Viral Transport System (VTM-RT) |
| Common Name | Transport culture medium |
| Review Panel | Microbiology |
| Classification Name andReference | 21 CFR 886.2390, Culture Media, Non-Propagating Transport |
| Device Product Code andPanel Code | JSM; Transport culture medium |
| Predicate Device | Copan Universal Transport Medium (UTM-RT) System (K042970) |
1. Intended Use and Indications for Use
iClean Viral Transport System (VTM-RT) is intended for the collection and transport of clinical specimens containing respiratory viruses, Chlamydiae, or Mycoplasma hominis from the collection site to the testing laboratory. The collection system is a culture based media that is intended to be used with standard laboratory examination, culture or with other assays that utilize stable recoverable infectious viral particles or bacteria.
2. Device Description
iClean Viral Transport System (VTM-RT) is intended for the collection and transport of clinical specimens containing respiratory viruses, Chlamydiae, or Mycoplasma hominis from the collection site to the testing laboratory. iClean Viral Transport System (VTM-RT) includes a plastic screw-cap tube with conical bottom containing 3mL transport medium. iClean Viral Transport System (VTM-RT) tubes can be supplied alone, or in a kit format with a flocked swab in a sterile peel pouch (or 100 kits packed together in a color box).
iClean Viral Transport System (VTM-RT) medium is stable at room temperature and consists of: Hanks balanced salt solution (HBSS), Bovine Serum Albumin (BSA), Gentamicin sulfate, Amphotericin B, Colistin, L-glutamic acid, HEPES buffer and Phenol red. The neutral environment constructed by Hank's buffer helps to increase the stability of the virus. Bovine Serum Albumin (BSA) acts as a protein stabilizer, forming a protective film on the protein shell of the virus, making it less likely to break down and ensuring the integrity of the virus. Gentamicin sulfate, amphotericin B, and colistin inhibit growth of bacteria or yeast. L-glutamic acid serves as an auxiliary energy source to keep cell and virus stability. HEPES buffer provides additional help to maintain a stable pH value environment thus increases the
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stability of virus. Phenol red is a pH indicator which serves as a visual quality control mechanism. As this medium does not contain cysteine, gelatin, or sucrose, it cannot be used to freeze virus samples for long-term preservation. The medium is isotonic and non-toxic to mammalian host cells.
3. Device Specification
iClean transport medium tubes can be supplied alone including a screw-cap tube containing 3mL of transport medium or in a kit format with one of the following collection swab configurations in a sterile peel pouch (or 100 kits packed together in a color box):
| Catalog No. | Description | Pack |
|---|---|---|
| CY-B-F005-20 | 3mL transport medium vial with iClean® nasopharyngealflocked Swab | Single pack or10*10 kits |
| CY-B-F005-20 | 3mL transport medium vial with iClean® buccal flockedSwab | Single pack or10*10 kits |
| CY-B-F005-20 | 3mL transport medium vial with iClean® oropharyngealflocked Swab | Single pack or10*10 kits |
| CY-B-F005-20 | 3mL transport medium vial with iClean® nasopharyngealflocked Swab and oropharyngeal flocked Swab | Single pack or10*10 kits |
The swabs in this kit are listed according to 21CFR Part 807 under the proprietary name of "Antiseptic Swab Applicator, Specimen Collection Flocked / Polyester / Foam / Cotton / Rayon Swab, Nasal / Oral / Throat / Cervical", registered under Huachenyang (Shenzhen) Technology Company Ltd (Registered establishment number: 3011649813). These swabs are not purchased in bulk, but are purchased in finished form, i.e., they are packaged, labeled, etc., consistent with their device listing criteria and status. Also, there were no recorded adverse events and/or quality problems of the swabs so far.
Media Formulation:
- Hanks balanced salt solution (HBSS) ●
- Bovine Serum Albumin (BSA) ●
- Gentamicin sulfate .
- . Amphotericin B
- Colistin
- L-glutamic acid .
- HEPES buffer ●
- Phenol red ●
iClean Viral Transport System (VTM-RT) should be stored in a clean, dry, ventilated environment at 2 - 25℃. The shelf-life of iClean Viral Transport System (VTM-RT) is 12 months after the manufacture date.
4. Principle of Operation
The iClean VTM-RT functions as a general transport medium kit for collecting and
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transporting clinical specimens. This transport medium is used to safely collect and transport viruses, Chlamydiae, or Mycoplasma hominis from collection sites to the testing laboratories. It is intended for use by Health Care Professionals (HCPs), the transport system allows for the collection of the specimen via the sterile swab, maintenance through a buffered media, prevention of microbial growth via antimicrobial agents, as well as a pH indicator. The media has been validated with culture recovery of virus. The sterile swab provided in the kit is packaged in peeled pouch for specimen collection. The cap from the vial is intended to be removed aseptically, and the sample collection swab is inserted into the vial containing the iClean VTM-RT Medium. After the collected sample is placed into the transport media, it is transported to the laboratory.
Proper specimen collection plays a critical role for successful isolation and identification of infectious organisms. Once the sample is collected it should be placed immediately into the media inside the tube and transported to the laboratory. Although the media can maintain even fragile organisms for long periods of time at room temperature, it is recommended that specimens be refrigerated at 2 - 8°C while in transit.
5. Substantial Equivalence
The iClean Viral Transport System (VTM-RT) is compared with the predicate device, Copan Universal Transport Medium (UTM-RT) System, in intended use, medium formulation, container, product configuration, shelf life, packaging and volume. The safety and effectiveness of the iClean Viral Transport System (VTM-RT) with the expanded claims is adequately supported by the substantial equivalence information, materials data, and testing results provided within this Premarket Notification. Below is a summary of comparison between iClean VTM-RT and predicate Copan Universal Transport Medium (UTM-RT) System:
| Subject Device(K212856) | Predicate Device(K042970) | |
|---|---|---|
| Device Trade Name | iClean Viral Transport System (VTM-RT kit) | Copan Universal Transport Medium(UTM-RT) System |
| Device Product Code andClassification | JSM, Class I | JSM, Class I |
| General Device Characteristic Similarities | ||
| Intended Use / IndicationsFor Use | iClean Viral Transport System(VTM- RT) is intended for thecollection and transport of clinicalspecimens containing respiratoryviruses, Chlamydiae, orMycoplasma hominis from thecollection site to the testinglaboratory. The collection system isa culture based media that isintended to be used with standardlaboratory examination, culture orwith other assays that utilize stablerecoverable infectious viral particlesor bacteria. | Copan Universal Transport Medium(UTM-RT) System is intended for thecollection and transport of clinicalspecimens containing viruses,Chlamydiae, Mycoplasma orUreaplasma from the collection site tothe testing laboratory. UTM-RT can beprocessed using standard clinicallaboratory operating procedures forviral, chlamydial, mycoplasma andureaplasma culture. |
| Storage Temperature | $20^{\circ}$ -25°C | Same |
| Tube Material | Plastic Screw-Cap Tube | Same |
| Single Use Device | Yes | Same |
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| pH | $7.4 \pm 0.4$ | Same |
|---|---|---|
| Shelf-life | 12 months | Same |
| Validation | Culture | Same |
| General Device Characteristic Differences | ||
| Media Formulation | • Hanks balanced salt solution (HBSS)• Bovine Serum Albumin (BSA)• Gentamicin sulfate• Amphotericin B• Colistin• L-Glutamic acid• HEPES buffer• Phenol red | • Hank's Balanced Salts solution (HBSS)• Bovine Serum Albumin (BSA)• Vancomycin• Amphotericin B• Colistin• L-Glutamic Acid• L-Cysteine• HEPES Buffer• Phenol Red• Gelatin Sucrose |
| Supported strains | • Adenovirus• Cytomegalovirus• Echovirus Type 30• Herpes Simplex Virus Type 1• Herpes Simplex Virus Type 2• Influenza A• Parainfluenza 3• Respiratory Syncytial Virus• Chlamydia pneumoniae• Chlamydia trachomatis• Mycoplasma hominis | • Adenovirus• Cytomegalovirus• Echovirus Type 30• Herpes Simplex Virus Type 1• Herpes Simplex Virus Type 2• Influenza A• Parainfluenza 3• Respiratory Syncytial Virus• Varicella Zoster Virus• Chlamydia pneumoniae• Chlamydia trachomatis• Mycoplasma hominis• Mycoplasma pneumoniae |
As seen with the comparison above, the differences between iClean Viral Transport System (VTM-RT) and the predicate's specification, safety and performance are comparable.
6. Shelf Life
The shelf life for the iClean VTM-RT was determined to be 12 months from the date of manufacture when stored in a clean, dry, ventilated environment at 20-25℃. The shelf life of the iClean VTM-RT was established using real-time aging performance test at time points T = 0, 1, 3, 6, 9 and 12 months. At each time point, appearance, volume, pH, antibiotic stability, and recovery study were assessed.
a. Appearance Inspection:
To evaluate appearance stability, three lots of iClean VTM were physically examined for real-time aging at timepoints T = 0, 1, 3, 6, 9, and 12 months. The media was stored in a clean, dry, ventilated environment at 20 - 25℃. At each time point, appearance of the product was inspected visually to be clear (i. e. no turbidity, no cloudy nor precipitation) and maintains a pink color (i.e., no color change from pink to yellow). Media volume was assessed to ensure each tube was filled to 3.0mL. All results were acceptable and support the claim that the VTM-RT is physically stable for 12 months.
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b. pH Stability
The pH of the media was used as one of the indicators to support product stability. The media was tested at time points T = 0, 1, 3, 6, 9 and 12 months after the manufacturing date. Three lots of VTM-RT media were stored in a clean, dry, ventilated environment under the recommended temperature conditions (20 - 25℃) and at the specified time intervals, 15 tubes from each of the three lots were removed from storage. The media inside each of the vials was evaluated using a calibrated pH meter. For all the tubes at each time point and each of the three lots, the pH was within the targe of 7.4 ± 0.4.
c. Antibiotic Stability:
iClean VTM-RT contains gentamicin sulfate, amphotericin B, and colistin to inhibit growth of bacteria or yeast. Stability of these antibiotics was evaluated through inoculating 5.0 x 10 - 4.5 x 10° CFU/mL of Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8099, and Candida albicans ATCC 10231 into iClean VTM media followed by appropriate storage and incubation. The inoculated media was tested immediately or stored in VTM for 24 hrs. Spiked media was transferred by pipetting 0.5 mL onto nutrient agar plates for S. aureus and E. coli and Sabouraud media for C. albicans. All plates evaluated at time T = 0 had 9.0 x 104 cfu/mL growth or greater while plates evaluated at 24 and 48 hrs. all had no growth.
d. Sterilization:
The iClean VTM kit is not claimed to be sterile nor is it intended to be sterilized by the end user. To decrease the chances of contamination the media uses specific manufacturing steps including sterilization of tubes and packaging by gamma radiation set at a dose of 5.3 Kilo Gray (KGy), in accordance with ISO 11137- 2:2015. The media is filtered using a 0.22 um sterile fiber membrane and then is aseptically filled into the pre-sterilized tubes. The aseptic status of the filtered iClean VTM-RT was then validated by a quality control process which evaluates the absence of growth of bacteria and fungi by spreading 0.1 mL of the filtered VTM- RT medium on nutrient agar and Sabouraud media plates and incubated at 35℃ ± 2℃ for 24 - 48 hours. No growth on any of the plates tested was observed.
The results collectively for appearance, volume, pH, antibiotic stability support the 12-month stability claim for the iClean VTM-RT.
7. Performance Testing - Recovery Studies
Performance of the iClean VTM was evaluated by Culture-Based Recovery Studies for viral and bacterial test strains. For Viral Recovery Studies, Fluorescent Foci Count method was utilized to evaluate the recovery of Influenza A (ATCC VR-544), Parainfluenza 3 (ATCC VR- 93) and Respiratory Syncytial Virus (ATCC VR-1401). This method was also utilized to evaluate the recovery of Chlamydia pneumoniae (ATCC VR-53592) and C. trachomatis (ATCC VR-880). For Bacterial Recovery Studies, Roll-Plate and Swab Elution Methods were utilized to evaluate the recovery of Mycoplasma hominis (ATCC VR-14027). Performance testing included nine lots of media. Each performance study summarized below used media lots that represents "New" for newly manufactured media, "Mid" for middle aged media (~5 months old) and "Old" for older media about to expire or recently expired media.
Viral Recovery Studies:
Virus stocks were diluted in pooled negative clinical matrix and each chosen dilution was inoculated
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into swab and placed into iClean VTM to store at 4℃ and 25℃ for 0, 24 and 48 hours respectively. For tissue culture, Hep-2 cells (ATCC CCL-23) or McCoy cells (ATCC CRL-1696) were grown to 95% confluency. When tissue culture plates were ready, 200 µl of each test samples were used to infect the monolayers and incubated. For detection, specific immunofluorescent antibody staining was used. The number of infectious particles were counted as Fluorescent Foci and calculated for each storage temperature and time points.
McCoy cell cultures were used for the recovery of Chlamydia (ATCC VR-53592) and C. trachomatis (ATCC VR-880). The results are presented in the Table 1 and Table 2 below. Any reduction in the foci count in the timepoints (0 to 48 hr.) was shown in percent decline.
| Test Strain | Lot No. | Lot Age | Average Recovery in Foci count/mL | Decline in 0 - 48 hrs. | ||
|---|---|---|---|---|---|---|
| Influenza A | 2021010105 | $7.1×10^3$ | $6×10^3$ | $4.9×10^3$ | 31% | |
| 2021010108 | Old | $1×10^4$ | $6.5×10^3$ | $5.7×10^3$ | 43% | |
| 2021010205 | $1.1×10^4$ | $8.8×10^3$ | $7×10^3$ | 36% | ||
| 2021060105 | $1.1×10^4$ | $9×10^3$ | $7.6×10^3$ | 31% | ||
| 2021060108 | Mid | $1.2×10^4$ | $8.6×10^3$ | $6.5×10^3$ | 46% | |
| 2021060201 | $8.6×10^3$ | $7.4×10^3$ | $4.8×10^3$ | 44% | ||
| 2021120101 | $9.2×10^3$ | $8.6×10^3$ | $6.7×10^3$ | 27% | ||
| 2021120102 | New | $1.2×10^4$ | $6.4×10^3$ | $8.8×10^3$ | 27% | |
| 2021120103 | $9.1×10^4$ | $8.5×10^3$ | $7.1×10^3$ | 92% | ||
| Parainfluenza 3 | 2021010105 | $8.2×10^3$ | $7.9×10^3$ | $8.6×10^3$ | -5% | |
| 2021010108 | Old | $7.9×10^3$ | $7×10^3$ | $7.1×10^3$ | 10% | |
| 2021010205 | $1.2×10^4$ | $7×10^3$ | $5×10^3$ | 58% | ||
| 2021060105 | $1×10^4$ | $8.4×10^3$ | $7.7×10^3$ | 23% | ||
| 2021060108 | Mid | $1.1×10^4$ | $8.2×10^3$ | $4.9×10^3$ | 55% | |
| 2021060201 | $1.4×10^4$ | $7.2×10^3$ | $6.2×10^3$ | 56% | ||
| 2021120101 | $1×10^4$ | $9.5×10^3$ | $5.5×10^3$ | 45% | ||
| 2021120102 | New | $1.1×10^4$ | $7.2×10^3$ | $8.4×10^3$ | 24% | |
| 2021120103 | $1.3×10^4$ | $1×10^4$ | $5.8×10^3$ | 55% | ||
| RespiratorySyncytial Virus | 2021010105 | $7.3×10^3$ | $8.5×10^3$ | $6.3×10^3$ | 14% | |
| 2021010108 | Old | $8.6×10^3$ | $8.4×10^3$ | $7.5×10^3$ | 13% | |
| 2021010205 | $1.2×10^4$ | $9.3×10^3$ | $7.8×10^3$ | 35% | ||
| 2021060105 | $1.1×10^4$ | $8.1×10^3$ | $6.3×10^3$ | 43% | ||
| 2021060108 | Mid | $1.1×10^4$ | $8.3×10^3$ | $7.4×10^3$ | 33% | |
| 2021060201 | $1.2×10^4$ | $8.9×10^3$ | $6.1×10^3$ | 49% | ||
| 2021120101 | $8.6×10^3$ | $1.1×10^4$ | $7.8×10^3$ | 9% | ||
| 2021120102 | New | $8.5×10^3$ | $1×10^4$ | $5.3×10^3$ | 38% | |
| 2021120103 | $8×10^3$ | $7.6×10^3$ | $6.2×10^3$ | 23% | ||
| Chlamydiapneumoniae | 2021010105 | $1.1\times10^6$ | $3.2\times10^5$ | $2.4\times10^5$ | 78% | |
| 2021010108 | Old | $2.8\times10^6$ | $1.0\times10^6$ | $5.3\times10^5$ | 81% | |
| 2021010205 | $2.6\times10^6$ | $1.5\times10^6$ | $3.1\times10^5$ | 88% | ||
| 2021060105 | Mid | $1.9\times10^6$ | $1.4\times10^6$ | $1.9\times10^5$ | 90% | |
| 2021060108 | $2.4\times10^6$ | $1.5\times10^6$ | $5.8\times10^5$ | 76% | ||
| 2021060201 | $1.8\times10^6$ | $5.1\times10^5$ | $5.7\times10^5$ | 68% | ||
| 2021120101 | $1.5\times10^6$ | $8.4\times10^5$ | $3.3\times10^5$ | 78% | ||
| 2021120102 | New | $1.9\times10^6$ | $1.4\times10^6$ | $3.3\times10^5$ | 83% | |
| 2021120103 | $1.2\times10^6$ | $1.0\times10^6$ | $8.1\times10^5$ | 33% | ||
| Chlamydiatrachomatis | 2021010105 | $1.6\times10^6$ | $4.6\times10^5$ | $5.0\times10^5$ | 69% | |
| 2021010108 | Old | $8.0\times10^5$ | $4.0\times10^5$ | $8.5\times10^5$ | -6% | |
| 2021010205 | $1.5\times10^6$ | $6.5\times10^5$ | $2.3\times10^5$ | 85% | ||
| 2021060105 | $2.3\times10^6$ | $5.3\times10^5$ | $3.2\times10^5$ | 86% | ||
| 2021060108 | Mid | $8.3\times10^5$ | $4.3\times10^5$ | $4.9\times10^5$ | 41% | |
| 2021060201 | $2.4\times10^6$ | $1.4\times10^6$ | $5.0\times10^5$ | 79% | ||
| 2021120101 | $1.2\times10^6$ | $9.2\times10^5$ | $6.6\times10^5$ | 45% | ||
| 2021120102 | New | $2.7\times10^6$ | $7.7\times10^5$ | $4.5\times10^5$ | 83% | |
| 2021120103 | $1.6\times10^6$ | $1.1\times10^6$ | $7.3\times10^5$ | 54% |
Table 1. Recovery of viruses and Chlamydiae at 4℃ storage.
{9}------------------------------------------------
Table 2. Recovery of viruses and Chlamydiae at 25°C storage.
| Test Strain | Lot No. | Lot Age | Average Recovery in Foci count/mL | Decline in0 - 48 hrs. | ||
|---|---|---|---|---|---|---|
| 0 hr. | 24 hrs. | 48 hrs. | ||||
| Influenza A | 2021010105 | $8.1\times10^3$ | $7.3\times10^3$ | $7\times10^2$ | 91% | |
| 2021010108 | Old | $9.3\times10^3$ | $7.2\times10^3$ | $1.1\times10^3$ | 88% | |
| 2021010205 | $1.1\times10^4$ | $7.2\times10^3$ | $1.7\times10^3$ | 85% | ||
| 2021060105 | $1.2\times10^4$ | $8.2\times10^3$ | $1.3\times10^3$ | 89% | ||
| 2021060108 | Mid | $1\times10^4$ | $8\times10^3$ | $2.1\times10^3$ | 79% | |
| 2021060201 | $9.4\times10^3$ | $5.6\times10^3$ | $1.9\times10^3$ | 80% | ||
| 2021120101 | $9.5\times10^3$ | $6.9\times10^3$ | $1.5\times10^3$ | 84% | ||
| 2021120102 | New | $1.1\times10^4$ | $6.7\times10^3$ | $2.2\times10^3$ | 80% | |
| 2021120103 | $1.2\times10^4$ | $7.7\times10^3$ | $1.4\times10^3$ | 88% | ||
| Parainfluenza 3 | 2021010105 | $1.3\times10^4$ | $6.1\times10^3$ | $1\times10^3$ | 92% | |
| 2021010108 | Old | $1.3\times10^4$ | $7.9\times10^3$ | $8.8\times10^2$ | 93% | |
| 2021010205 | $1.1\times10^4$ | $8.7\times10^3$ | $9.1\times10^2$ | 92% | ||
| 2021060105 | $9.3\times10^3$ | $5.9\times10^3$ | $1.9\times10^3$ | 80% | ||
| 2021060108 | Mid | $9.2\times10^3$ | $8.1\times10^3$ | $8.4\times10^2$ | 91% | |
| 2021060201 | $9.5\times10^3$ | $7.4\times10^3$ | $7\times10^2$ | 93% | ||
| 2021120101 | $1\times10^4$ | $9.1\times10^3$ | $2.1\times10^3$ | 79% | ||
| 2021120102 | New | $1.2\times10^4$ | $8.6\times10^3$ | $1.7\times10^3$ | 86% | |
| 2021120103 | $1.1\times10^4$ | $7.3\times10^3$ | $1.6\times10^3$ | 85% |
{10}------------------------------------------------
| RespiratorySyncytial Virus | ||||||
|---|---|---|---|---|---|---|
| 2021010105 | $9.2\times10^3$ | $7.8\times10^3$ | $1.9\times10^3$ | 79% | ||
| 2021010108 | Old | $8\times10^3$ | $9.4\times10^3$ | $1.1\times10^3$ | 86% | |
| 2021010205 | $1.1\times10^4$ | $5.6\times10^3$ | $9.5\times10^2$ | 91% | ||
| 2021060105 | $1.3\times10^4$ | $6\times10^3$ | $6.8\times10^2$ | 95% | ||
| 2021060108 | Mid | $8.7\times10^3$ | $6.4\times10^3$ | $5.9\times10^2$ | 93% | |
| 2021060201 | $1.2\times10^4$ | $8.7\times10^3$ | $1.3\times10^3$ | 89% | ||
| 2021120101 | $8.4\times10^3$ | $5.9\times10^3$ | $6.3\times10^2$ | 93% | ||
| 2021120102 | New | $1\times10^4$ | $7.8\times10^3$ | $1.1\times10^3$ | 89% | |
| 2021120103 | $9.6\times10^3$ | $6.5\times10^3$ | $1.3\times10^3$ | 86% | ||
| Chlamydiapneumoniae | ||||||
| 2021010105 | $1.5\times10^6$ | $6.6\times10^5$ | $1.9\times10^5$ | 87% | ||
| 2021010108 | Old | $8.5\times10^5$ | $4.2\times10^5$ | $2.5\times10^6$ | -194% | |
| 2021010205 | $1.4\times10^6$ | $1.1\times10^6$ | $2.3\times10^5$ | 84% | ||
| 2021060105 | $1.8\times10^6$ | $8.3\times10^5$ | $1.8\times10^5$ | 90% | ||
| 2021060108 | Mid | $2.7\times10^6$ | $9.1\times10^5$ | $1.9\times10^5$ | 93% | |
| 2021060201 | $1.4\times10^6$ | $1.0\times10^6$ | $2.2\times10^5$ | 84% | ||
| 2021120101 | $1.7\times10^6$ | $3.0\times10^5$ | $2.4\times10^5$ | 86% | ||
| 2021120102 | New | $1.3\times10^6$ | $9.1\times10^5$ | $7.9\times10^4$ | 94% | |
| 2021120103 | $1.7\times10^6$ | $8.5\times10^5$ | $2.1\times10^5$ | 88% | ||
| Chlamydiatrachomatis | ||||||
| 2021010105 | $1.2\times10^6$ | $6.7\times10^5$ | $1.7\times10^5$ | 86% | ||
| 2021010108 | Old | $2.6\times10^6$ | $4.8\times10^5$ | $1.7\times10^5$ | 93% | |
| 2021010205 | $2.2\times10^6$ | $4.3\times10^5$ | $9.6\times10^4$ | 96% | ||
| 2021060105 | $1.5\times10^6$ | $4.7\times10^5$ | $2.4\times10^5$ | 84% | ||
| 2021060108 | Mid | $9.2\times10^5$ | $3.4\times10^5$ | $1.5\times10^5$ | 84% | |
| 2021060201 | $1.3\times10^6$ | $7.7\times10^6$ | $7.5\times10^4$ | 42% | ||
| 2021120101 | $1.1\times10^6$ | $1.0\times10^6$ | $7.2\times10^4$ | 35% | ||
| 2021120102 | New | $2.4\times10^6$ | $9.0\times10^5$ | $2.0\times10^5$ | 92% | |
| 2021120103 | $1.7\times10^6$ | $1.0\times10^6$ | $1.8\times10^5$ | 89% |
Bacterial Recovery Studies:
Performance of iClean VTM for bacterial recovery was determined using roll plate swab elution methods. Both the roll plate and swab elution studies follow the FDA recognized sections of CLSI M40-A2:2014 Quality Control of Microbiological Transport Systems: Approved Standard – Second Edition.
Roll-plate method:
For the roll-plate method, Mycoplasma hominis suspensions were prepared to approximately 0.5 McFarland standard (1.5 × 10° CFU/mL) in 0.85% physiological saline followed by 10- fold serial dilutions in pooled negative matrix. Swabs in triplicate were spiked with 100 uL of each dilution and placed in the iClean VTM and maintained under refrigerated or at room temperature. After 0, 24 and 48 hours, the swabs were removed and rolled directly onto agar plates which were incubated to grow colonies. Average CFU/roll-plate calculated for each timepoints were presented in Table 4. According to CLSI M40 A2 guidelines, the inoculum dilutions yielding time-zero plates with closely approaching 300 CFU was used to complete the viability studies. The acceptance criteria were set to a recovery of ≥ 10 CFU following the specified maintenance time at the iClean VTM.
{11}------------------------------------------------
| Test Strain | Lot No. | Lot Age | Average Recovery in CFU/roll-plate | 0 hr. | 24 hrs. | 48 hrs. |
|---|---|---|---|---|---|---|
| Mycoplasma hominis(ATCC VR-14027) | 2021010105 | Old | 275 | 233 | 150 | |
| 2021010108 | 277 | 252 | 165 | |||
| 2021010205 | 264 | 220 | 118 | |||
| 2021060105 | Mid | 267 | 248 | 139 | ||
| 2021060108 | 283 | 240 | 158 | |||
| 2021060201 | 279 | 250 | 95 | |||
| 2021120101 | New | 252 | 234 | 100 | ||
| 2021120102 | 292 | 289 | 181 | |||
| 2021120103 | 248 | 212 | 111 |
| Table 3. Roll-Plate Method of recovery for storage at refrigerated conditions (2-8℃). |
|---|
Table 4. Roll-Plate Method of recovery for storage at room temperature (22-25°C).
| Test Strain | Lot No. | Lot Age | Average Recovery in CFU/Roll-plate | ||
|---|---|---|---|---|---|
| 0 hr. | 24 hrs. | 48 hrs. | |||
| Mycoplasma hominis(ATCC VR-14027) | 2021010105 | 267 | 283 | 107 | |
| 2021010108 | Old | 323 | 227 | 94 | |
| 2021010205 | 310 | 282 | 112 | ||
| Mycoplasma hominis(ATCC VR-14027) | 2021060105 | Mid | 261 | 207 | 92 |
| 2021060108 | 276 | 246 | 130 | ||
| 2021060201 | 276 | 215 | 108 | ||
| Mycoplasma hominis(ATCC VR-14027) | 2021120101 | New | 320 | 251 | 72 |
| 2021120102 | 297 | 278 | 82 | ||
| 2021120103 | 306 | 247 | 134 |
Swab Elution Method:
For the swab elution method, the Mycoplasma hominis inocula were prepared in a manner similar to that for the roll-plate method. The initial bacterial suspensions were diluted by 10-4 and dispensed 100 uL onto swabs in triplicate. The swabs were then placed in the iClean VTM and maintained under refrigerated or at room temperature for the specified timepoints. After 0, 24, and 48 hours the swabs were removed, and 10-fold serial dilutions were prepared. From each of the dilution, 50 µL was dispensed onto the agar plate and incubated to allow the growth of colonies. The results calculated in average CFU/mL for the specified time points are presented in Table 5 and 6. According to CLSI document M40-A2, the acceptance criteria for viability in the swab elution method was considered to be no more than a 3 log10 change in CFU count between the zero-time and the 48 hours - time points.
Table 5. Swab Elution Method of recovery for storage at refrigerated conditions (2-8°C).
| Test Strain | Lot No. | Lot Age | Average Recovery in CFU/mL | Change in log10 (0 to 48 hrs.) | ||
|---|---|---|---|---|---|---|
| 0 hr. | 24 hrs. | 48 hrs. | ||||
| 2021010105 | $2.1\times10^6$ | $5.1\times10^5$ | $2.1\times10^5$ | -1.0 | ||
| 2021010108 | Old | $1.3\times10^6$ | $7.2\times10^5$ | $3.3\times10^5$ | -0.6 | |
| 2021010205 | $1.4\times10^6$ | $3.4\times10^5$ | $1.4\times10^5$ | -1.0 | ||
| Mycoplasma hominis | 2021060105 | $1.5\times10^6$ | $7.2\times10^5$ | $2.5\times10^5$ | -0.8 | |
| 2021060108 | $2.3\times10^6$ | $4.1\times10^5$ | $2.2\times10^5$ | -1.0 |
{12}------------------------------------------------
| (ATCC VR-14027) | 2021060201 | Mid | $1.9\times10^6$ | $5.8\times10^5$ | $2.6\times10^5$ | -0.9 |
|---|---|---|---|---|---|---|
| 2021120101 | $8.9\times10^5$ | $4.9\times10^5$ | $1.8\times10^5$ | -0.7 | ||
| 2021120102 | New | $1.1\times10^6$ | $8.7\times10^5$ | $2.9\times10^5$ | -0.6 | |
| 2021120103 | $1.9\times10^6$ | $1.0\times10^6$ | $6.4\times10^5$ | -0.5 |
Table 6. Swab Elution Method of recovery for storage at room temperature (22-25°C).
| Test Strain | Lot No. | LotAge | Average Recovery inCFU/mL | Change in log10(0 to 48 hrs.) | ||
|---|---|---|---|---|---|---|
| 0 hr. | 24 hrs. | 48 hrs. | ||||
| Mycoplasma hominis(ATCC VR-14027) | 2021010105 | Old | 2.7×106 | 8.9×105 | 1.4×105 | -1.3 |
| 2021010108 | 2.1×106 | 1.1×106 | 2.2×105 | |||
| 2021010205 | 1.8×106 | 3.5×105 | 8.6×104 | |||
| 2021060105 | Mid | 8.4×105 | 7.0×105 | 1.8×105 | -0.7 | |
| 2021060108 | 1.2×106 | 5.7×105 | 1.6×105 | |||
| 2021060201 | 2.0×106 | 8.9×105 | 1.1×105 | |||
| 2021120101 | New | 1.7×106 | 3.9×105 | 2.1×105 | -0.9 | |
| 2021120102 | 2.7×106 | 8.7×105 | 1.0×105 | |||
| 2021120103 | 2.2×106 | 2.8×105 | 9.4×104 |
8. Conclusion of the Culture-based Recovery Studies
The iClean VTM demonstrated the recovery of tested viruses, Chlamydia pneumoniae, C. trachomatis and Mycoplasma hominis in all replicates at tested incubation times and storage conditions.
§ 866.2390 Transport culture medium.
(a)
Identification. A transport culture medium is a device that consists of a semisolid, usually non-nutrient, medium that maintains the viability of suspected pathogens contained in patient specimens while in transit from the specimen collection area to the laboratory. The device aids in the diagnosis of disease caused by pathogenic microorganisms and also provides epidemiological information on these diseases.(b)
Classification. Class I (general controls).