(1057 days)
The SMART Leish is a qualitative diagnostic real-time PCR test for the rapid detection of Leishmania species and the identification of L. major in skin lesion scrapings and punch biopsies from individuals suspected of having cutaneous leishmaniasis. The test utilizes real-time polymerase chain reaction assay on the Cepheid SmartCycler® II Dx to detect Leishmania species and L. major. The SMART Leish is indicated for use in patients with clinical presentations and travel history.
The SMART Leish consists of an assay reagent kit and assay definition files for the polymerase chain reaction (PCR) instrument platform. The kit contains sufficient reagents, in lyophilized bead form, to qualitatively assay 50 clinical samples for both Leishmania genus and L. major targets. Additional required accessories that are specified include the PCR instrument platform, a deoxyribonucleic acid (DNA) purification kit, and a positive extraction control. The device components and required accessories are listed as follows.
For the Leishmania assays, a tissue specimen (skin scraping or punch biopsy) from an individual suspected of being infected with Leishmania species or L. major is collected in 70% or 100% ethanol, and the DNA is extracted from the specimen using the Qiagen QlAamp DNA Mini Kit. An aliquot of this DNA is tested using the Leishmania genus assay, which will amplify a portion of DNA encoding the Leishmania species 16S ribosomal ribonucleic acid (rRNA) gene if present. Amplified targets are detected using a TaqMan® hybridization probe with 6-carboxyfluorescein.(FAM) reporter dye (517 nm) and a Black Hole Quencher® (BHQ). This assay also contains a positive internal control consisting of a nonsensical, non-naturally occurring DNA sequence, with Texas Red reporter dye (615 nm) and BHQ, used to detect evidence of PCR inhibition and confirm the integrity of assay reagents in negative specimens. If the sample tests positive for Leishmania species, another aliquot of DNA may be tested using the L. major assay, which will amplify a portion of DNA encoding the GPI gene that is specific to L. major. Amplified targets are detected using a TaqMan hybridization probe with FAM reporter dye and a BHQ. The DNA amplification, detection of fluorescence, and interpretation of signals are done automatically by the SmartCycler instrument for each assay. The thermocycling protocols take approximately 45-55 minutes.
Here's a breakdown of the acceptance criteria and the study details for the SMART Leish device, based on the provided text:
Acceptance Criteria and Reported Device Performance
The document does not explicitly state pre-defined "acceptance criteria" in terms of numerical thresholds for sensitivity, specificity, or agreement. However, the reported performance suggests what the submitter considered acceptable for demonstrating substantial equivalence. The provided data focuses on the clinical performance and various analytical performance metrics.
| Performance Metric | Acceptance Criteria (Inferred) | Reported Device Performance (Leishmania Genus Assay) | Reported Device Performance (L. major Assay) |
|---|---|---|---|
| Clinical Performance | Demonstrated effectiveness in detecting Leishmania species and identifying L. major in suspected cutaneous leishmaniasis cases. | Sensitivity: 223/225 = 99.1%Specificity: 58/87 = 66.7% | Sensitivity: 92/96 = 95.8%Specificity: 83/91 = 91.2% |
| Analytical Reactivity/Inclusivity | Robust positive results with target Leishmania strains. | 100% positive results for 36 Leishmania strains. | 100% positive results for 11 L. major strains. |
| Analytical Specificity (Cross-Reactivity) | No significant cross-reactivity with non-target organisms. | 98.3% (2 false positives out of 85 non-target samples). Minimal cross-reactivity with C. fasciculata. | 100% (0 false positives out of 25 non-L. major samples). |
| Analytical Sensitivity (LOD) | Lowest detectable amount of DNA/parasites with ≥95% consistency. | Assay LOD: 0.14 pgeLOD: 250 parasites | Assay LOD: 2.1 pgeLOD: 1,000 parasites |
| Reproducibility | Consistent results across different laboratories, operators, and days, for various concentrations of purified DNA, cultured parasites, and mock human samples. | Overall Agreement for Mock Human Samples: 98.6% (71/72) | Overall Agreement for Mock Human Samples: 100% (41/41) |
| Interfering Substances | Performance not significantly affected by common PCR inhibitors at clinically relevant concentrations. | See Table 10 for specific concentrations and non-inhibition limits. | Not explicitly detailed for L. major assay in this section. |
Study Details
2. Sample size used for the test set and the data provenance
- Leishmania Genus Assay: 312 prospective specimens (225 positive, 87 negative)
- Leishmania major Assay: 187 prospective specimens (96 positive, 91 negative)
- Data Provenance: Multi-center, retrospective clinical study conducted at two military U.S. hospital sites. The patient population consisted entirely of military personnel who were at the time, or had previously been, deployed to Southwest Asia—primarily Afghanistan and Iraq.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts
The document does not specify the number or qualifications of experts used to establish the ground truth.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
The document does not specify an adjudication method. The ground truth was established by "culture and/or microscopy." In cases of discrepancy (false positives by SMART Leish), sequencing methods were used for further investigation, but this is a reconciliation rather than a primary adjudication method for establishing initial ground truth.
5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
- No, an MRMC comparative effectiveness study was not done.
- This device is a diagnostic assay (PCR test), not an AI-assisted imaging or interpretation tool for human readers. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not apply here.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
- Yes, a standalone performance study was done. The SMART Leish is a fully automated diagnostic assay where "DNA amplification, detection of fluorescence, and interpretation of signals are done automatically by the SmartCycler instrument for each assay." The clinical performance data presented (Tables 2 and 3) reflects this standalone performance against the clinical truth.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
The ground truth for the clinical performance study (test set) was established using culture and/or microscopy. For discordant results (false positives by SMART Leish compared to initial ground truth), sequencing methods were used for further analysis.
8. The sample size for the training set
The document does not specify a "training set" or its size. As a PCR diagnostic assay, it would typically undergo a development and optimization phase, but the document focuses on validation/testing. The "analytical reactivity" and "analytical specificity" studies used various Leishmania strains and non-target organisms, which could be considered part of the development/validation process.
- Analytical Reactivity Study: 36 Leishmania strains.
- Analytical Specificity Study: 121 DNA samples (11 L. major, 25 non-L. major, 85 non-target organisms from various categories).
9. How the ground truth for the training set was established
Since no explicit "training set" is mentioned in the context of machine learning, the establishment of ground truth would refer to the characteristics of the strains used in analytical studies. These would be:
- For Leishmania strains: Identity established through standard microbiological and molecular characterization methods (e.g., taxonomic classification, phenotypic and genotypic analysis, likely confirmed by sequencing).
- For non-target organisms: Identity established through standard microbiological and molecular characterization methods.
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510 (k) Summary - K081868
MAY 25 2011
Submitter
Point of Contact
Alternate Contact Person
Date Summary Prepared
Trade/Proprietary Name Common or Usual Name
Classification Name
Regulation Number
Product Code
Review Panel
Classification
Predicate Device
Department of the Army U.S. Army Medical Research and Materiel Command 1430 Veterans Drive Fort Detrick, MD 21702-5012
Dr. Robert E. Miller, Ph.D., RAC Director, Division of Regulated Activities and Compliance 301-619-0317
Patricia M. Beverly, MT,RAC Regulatory Scientist 301-619-2980 patricia.m.beverly@us.army.mil or 301-619-1087 usammdaregulatoryaffairs@amedd.army.mil
April 28, 2011
SMART Leish
Leishmania Real-time Polymerase Chain Reaction Diagnostic Assay,
Trypanosoma spp. serological reagents
866.3870
OUZ
Microbiology 83
Class I
1
Amizyme-Leishmania spp. Test Kit
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Intended Use
The SMART Leish is a qualitative diagnostic real-time PCR test for the detection of Leishmania species and the identification of L. major in skin lesion scrapings and punch biopsies from individuals suspected of having cutaneous leishmaniasis. The test utilizes real-time polymerase chain reaction assay on the Cepheid SmartCycler® II Dx to detect Leishmania species and L. major.
WARNING
The SMART Leish is intended for use only by trained laboratory personnel in Department of Defense laboratories. Clinical performance has not been established with strains other than L. major.
Device Description
The SMART Leish consists of an assay reagent kit and assay definition files for the polymerase chain reaction (PCR) instrument platform. The kit contains sufficient reagents, in Iyophilized bead form, to qualitatively assay 50 clinical samples for both Leishmania genus and L. major targets. Additional required accessories that are specified include the PCR instrument platform, a deoxyribonucleic acid (DNA) purification kit, and a positive extraction control. The device components and required accessories are listed as follows.
For the Leishmania assays, a tissue specimen (skin scraping or punch biopsy) from an individual suspected of being infected with Leishmania species or L. major is collected in 70% or 100% ethanol, and the DNA is extracted from the specimen using the Qiagen QlAamp DNA Mini Kit. An aliquot of this DNA is tested using the Leishmania genus assay, which will amplify a portion of DNA encoding the Leishmania species 16S ribosomal ribonucleic acid (rRNA) gene if present. Amplified targets are detected using a TaqMan® hybridization probe with 6-carboxyfluorescein.(FAM) reporter dye (517 nm) and a Black Hole Quencher® (BHQ). This assay also contains a positive internal control consisting of a nonsensical, non-naturally occurring DNA sequence, with Texas Red reporter dye (615 nm) and BHQ, used to detect evidence of PCR inhibition and confirm the integrity of assay reagents in negative specimens. If the sample tests positive for Leishmania species, another aliquot of DNA may be tested using the L. major assay, which will amplify a portion of DNA encoding the GPI gene that is specific to L. major. Amplified targets are detected using a TaqMan hybridization probe with FAM reporter dye and a BHQ. The DNA amplification, detection of fluorescence, and interpretation of signals are done automatically by the SmartCycler instrument for each assay. The thermocycling protocols take approximately 45-55 minutes.
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Substantial Equivalence Information
-
- Predicate device name: Amizyme-Leishmania spp. Test Kit
-
- 510k number : K842526
Comparison with predicate:
| Device Aspect | SMART Leish | Reference Device |
|---|---|---|
| Intended Use | Detection of Leishmania spp | Detection of Leishmania spp |
| Assay Type | Qualitative | Qualitative |
| Agent measured | Leishmania genus and L. major DNA | Leishmania spp |
| Technology | Qiagen QIAap DNA Mini Kit | None |
| Controls | External - positive and negative controlsfor both the extraction process anddetection assay runsInternal - positive control for PCRinhibitors in sample | External positive and negativecontrols |
| Differences | ||
| Instrument Platform | Cepheid SmartCycler | None |
| Automation | Automated DNA amplification cycling,probe detection, and interpretation ofresults | Manual method |
| Sample Collection | Skin punch biopsy or scrapings, stored inethanol | Serum |
| Endpoint detection method | PCR amplification of DNA sequencesunique to the organisms of interest withdetection by hybridization probesincorporating reporter dyes | Indirect Immunofluoresence (IFA)Fluorescent conjugate secondaryantibodies on a prepared slide |
Table 1. Similarities
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Clinical Performance Testing
The clinical performance of the SMART Leish assay was established in a multi-center, retrospective clinical study conducted at two military U.S. hospital sites.
A total of 312 Leishmania genus and 187 Leishmania major prospective specimens were collected from adults 18 years or older and evaluated in the SMART Leish assay and compared to culture and/or microscopy. Evaluated specimens were skin lesion scrapings and punch biopsies from individuals suspected of having cutaneous leishmaniasis. The patient population consisted entirely of military personnel who were at the time, or had previously been, deployed to Southwest Asia—primarily Afghanistan and Iraq. SMART Leish performance versus culture and microcopy, including 95% confidence intervals, is detailed below.
| Clinical Truth | ||||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| SMART Leish -LeishmaniaGenus Assay | Positive | 223 | 29 | 252 |
| Negative | 2 | 58 | 60 | |
| Total | 225 | 87 | 312 |
Table 2. Summary of SMART Leish Combined Studies for the Leishmania genus Assay
*Diagnosis of leishmaniasis by culture and slide techniques results in lower clinical sensitivity than PCR assays. In this clinical data set there were 29 false positives generated for the Leishmania Genus assay. Twenty seven of these Genus false positives were determined to be Genus positive using sequencing methods.
| Table 3. Summary of SMART Leish Combined Studies for the Leishmania major Assay | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| -- | -- | -- | -- | -- | -- | -- | --------------------------------------------------------------------------------- | -- | -- |
| Clinical Truth | ||||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| SMART Leish - L.major Assay | Positive | 92 | 8 | 100 |
| SMART Leish - L.major Assay | Negative | 4 | 83 | 87 |
| SMART Leish - L.major Assay | Total | 96 | 91 | 187 |
Diagnosis of leishmaniasis by culture and slide techniques results in lower clinical sensitivity than PCR assays. In this clinical data set there were eight false positives generated for the L. major assay, of which three of were determined to be L. major using sequencing methods.
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ANALYTICAL PERFORMANCE
Analytical Reactivity Study
Two separate studies were done with Leishmania genus and Leishmania major assays using 36 Leishmania strains. The goal was to determine whether the assays cross-reacts with the nucleic acids from non-target organisms. In the Leishmania genus study, DNA from 36 Leishmania strains was tested and gave robust positive results with the Leishmania genus PCR assay. In the Leishmania major study, 11 of 36 strains were Leishmania major species. All 11 tested gave robust positive results with the Leishmania major PCR assay. None of the 25 non-Leishmania major samples tested positive. See Table 4.
Table 4. List of Leishmania Organisms Tested in Reactivity Study
| Leishmania major (11) | Leishmania mexicana |
|---|---|
| Leishmania tropica (9) | Leishmania amazonensis |
| Leishmania infantum (7) | Leishmania peruviana |
| Leishmania donovani (2) | Leishmania guyanensis (2) |
| Leishmania braziliensis | Leishmania tarentolae |
Analytical Specificity
A total of 121 DNA samples were evaluated in the specificity study. The DNA from 11 Leishmania major strains tested in this study gave robust positive results with the Leishmania major PCR assay. Twenty-five non-Leishmania major samples tested negative; hence the analytical specificity was 100%. The percentage of false positives was 0%, and the percentage of false negatives is 0%. In the Leishmania genus study, 85 of the 121 samples were non-target DNA samples. Eighty-five were negative, 2 false positives and 36 were positive. The analytical specificity of the Leishmania genus PCR assay was 98.3%, false positive rate was 2.4% and the false negative rate was 0%. See Table 5 for list of organisms used in the study.
Nucleic acids from non-target organisms were tested for both the Leishmania genus and Leishmania major assays. DNA samples with concentrations used in this study included purified DNA from bacteria (52; conc =50 - 46,250 pg/uL), fungi (11; conc= 50,000 - 97,000 pg/uL ), viruses (7; conc= 5,950 – 30,990 pg/uL), mammals (3; conc = 20 pg/uL (human, bovine, and murine)), human melanoma cell lines (3; conc = 10,200 – 25,200 pg/uL), Leishmania major (11; conc = 21,540 = 83,000 pg/uL), 25 additional Leishmania species (not-L. major) (conc = 16,680 – 592,00 pg/uL), and 9 trypanosomes representing 6 species (conc = 12,760 – 95,000 pg/uL).
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Table 5. List of Organisms Used in Specificity Study
Bacteria
Acinetobacter baumanni Bacillus anthracis (3) Bacillus cereus (2) Bacillus subtilis var. niger Bacillus thuringiensis Brucella abortus Brucella melitiensis Brucella suis Clostridium botulinum type A Clostridium botulinum type B Clostridium perfringens (2) Clostridium sordelli Enterobacter aerogenes Enterococcus durans Enterococcus faecalis Escherichia coli Francisella tularensis (2) Haemophilus influenzae Klebsiella oxytoca Klebsiella pneumoniae Moraxella cattaharalis Neisseria lactamica Pasteurella multocida Proteus mirabilis Proteus vulgaris Providencia stuartii Pseudomonas aeruginosa Staphylococcus aureus (4)
Staphylococcus hominis Stenotrophomonas maltophilia Streptococcus pyogenes (2) Streptococcus sp. (B) Streptococcus (F2) Yesinia nestis Yersinia
Mycobacteria Mycobacteria abscessus Mycobacteria fortuitum Mycobacteria marinum Mycobacteria spp Mycobacteria tuberculosis Mycobacteria ulcerans
Viruses
Human Papilloma Virus Herpes Simplex Virus Type I Herpes Simplex Virus Type II (2) Varicella Zoster Virus (2)
Fungi Microsporium gypseum Cladophialophora carrionii Fonsecaea pedrosoi
Rhinocladiella compacta Phialophora verrucosa Trichophyton tonsurans Trichophyton mentagrophytes Trichophyton soudanense Arthroderma benhamiae Sporothrix schenckii Microsporum canis
Mammalian Bovine Human Murine Human melanoma cell line (3)
Trypanosoma
Trypanosoma cruzi (2) Trypanosoma rhodesiense Trypanosoma rangeli (2) Trypanosomal lewisi lincicome Trypanosoma brucei gambiense Crithidia fasciculata (2)
Two strains of C. fasciculata were determined to be low-level cross-reactors based on falsepositive results observed. The degree of cross-reactivity of the Leishmania genus assay for C. fasciculata can be considered low level or weak. No cross-reactors were determined for the L. major assay through the study.
Supplementary in silico analysis was done comparing the Smart Leish primer and probe sequences to sequences from the following organisms that could potentially result in a clinical presentation similar to cutaneous leishmaniasis: Corynebacterium diphtheria (veldt sores), Mycobacteria (Tropical Ulcer), Mycobacterium tuberculosis (Lupus vulgaris), Treponema pallidum (Tertiary Syphilis), Treponema pertenue (Yaws), and Blastomyces (Blastomycosis). Each Smart Leish primer and probe contained at least four mismatches to those sequences. Based on these alignments, no cross-reactivity was predicted with the Smart Leish primers and probes.
Analytical Sensitivity
The assay linearity of SMART Leish for Leishmainia genus and Leishmania major was determined using eight different concentrations of purified L. major DNA tested in a 10-fold dilution series from 14.26 ng to 0.001426 pg and 21.39 ng to 0.002139 pg, respectively, DNA per reaction. Based on these results, additional studies were done to determine the analytical sensitivity (limit of detection or LOD) which was defined as the lowest amount of purified L. major DNA that consistently provided positive test results 95% of the time.
7
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In addition to the LOD determination, six different concentrations of purified L. major parasites were tested in a 10-fold dilution of 5 X 102 parasites/mL (equivalent to 107 to 102 parasites/extraction) to determine the minimum number of L. major parasites that can be taken through the extraction and testing procedure and still give a positive result for the SMART Leish assays (extraction LOD).
Table 6 summarizes the results of the assay LOD for the L. genus and L. major assays, and Table 7 summarizes the results of the extraction LOD (eLOD) study.
Table 6. Summary of the Measuring Range and LOD for the Leishmania genus and L. major Assays (Values are per assay reaction)
| Assay | Linear Range1 | Assay LOD1.2 | LOD Equates to |
|---|---|---|---|
| Leishmania Genus Assay | 0.014-14,260 pg | 0.14 pg | 4 genome copies with~200 copies/genome oftarget gene |
| L. major Assay | 0.2139-21,390 pg | 2.1 pg | 62 genome copies |
'Amount of L. major DNA in the PCR reaction. LOD refers to the minimum amount of L. major DNA that can be put into a SMART Leish PCR reaction and still result in a positive SMART Leish test greater than or equal to 95% of the time.
295% accuracy across different operators and instruments and from multiple L. major strains
Table 7. Summary of Assay eLOD for the Leishmania genus and L. major Assays Using Parasite Samples. Values are per extraction.
| Assay | eLOD¹ | LOD Equates to |
|---|---|---|
| Leishmania Genus Assay | 250 parasites | ~5 genome copies at assayreaction stage |
| L. major Assay | 1,000 parasites | ~30 genome copies at assayreaction stage |
195% accuracy across different operators and days and from multiple L. major strains
- Extraction LOD refers to the minimum number of L. major parasites that can be taken through the Qiagen extraction procedure and still consistently result in a positive SMART Leish test greater than or equal to 95% of the time. Extraction LOD was determined across different operators, different days, and using multiple L. major strains.
Reproducibility
Precision was evaluated internally at the study sites. The following tables summarize the between laboratory reproducibility results for the Leishmania genus and L. major assays, respectively. These tables include the results of the studies done with purified DNA, cultured parasites, and mock human samples. The negative mock human samples were not further tested using the L. major assay if the initial Leishmania genus assay result was negative.
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Table 8. Summary of Between Laboratory Reproducibility Results for the Leishmania Genus Assay ﺍﻟﻘﺎﺩﺭ
| SampleType | Sample ID | Concentration | Number ofReplicates(over 5 days) | LaboratoryA(WRAIR) | LaboratoryB(BAMC) | LaboratoryC(MAMC) | TotalAgreement | % TotalAgreement |
|---|---|---|---|---|---|---|---|---|
| genomeequivalents/µL | ||||||||
| PurifiedDNA | Low | 2.0 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% |
| Medium | 20 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| High | 200 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| PositiveAgreementfor PurifiedDNA | 180/180 | 100% | ||||||
| (parasites/extraction) | ||||||||
| CulturedParasites | Low | 1,000 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% |
| Medium | 10,000 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| High | 25,000 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| PositiveAgreementfor CulturedParasites | 180/180 | 100% | ||||||
| (parasites/extraction) | ||||||||
| MockHumanSamples | Negative | 0 | 10 | 10/10 | 10/10 | 10/10 | 30/30 | 100% |
| Positive | 1.1x10^6 | 14 | 14/14 | 13/14 | 14/14 | 41/42 | 97.6% | |
| OverallAgreementfor MockHumanSamples | 71/72 | 98.6% |
SMART LEISH 510(k) Summary
:
:
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Table 9. Summary of between Laboratory Reproducibility Results for the Leishmania major Assay
| SampleType | Sample ID | Concentration | Number ofReplicates(over 5days) | LaboratoryA(WRAIR) | LaboratoryB(BAMC) | LaboratoryC(MAMC) | TotalAgreement | % TotalAgreement |
|---|---|---|---|---|---|---|---|---|
| genomeequivalents/µL | ||||||||
| PurifiedDNA | Low | 2.0 | 20 | 15/20 | 7/20 | 15/20 | 37/60 | 61% |
| Medium | 20 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| High | 200 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| PositiveAgreementfor PurifiedDNA | 157/180 | 87.2% | ||||||
| (parasites/extraction) | ||||||||
| CulturedParasites | Low | 1,000 | 20 | 19/20 | 20/20 | 16/20 | 55/60 | 91.6% |
| Medium | 10,000 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| High | 25,000 | 20 | 20/20 | 20/20 | 20/20 | 60/60 | 100% | |
| PositiveAgreementforCulturedParasites | 175/180 | 97.2% CI | ||||||
| (parasites/ | ||||||||
| extraction) | ||||||||
| MockHumanSamples | Negative | 0 | 0/0 | 0/0 | 0/0 | 0/0 | 0/0 | 100% |
| Positive | $1.1x10^6$ | 14 | 14/14 | 13/13 | 14/14 | 41/41 | 100% | |
| OverallAgreementfor MockHumanSamples | 41/41 | 100% |
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INTERFERING SUBSTANCES
The Leishmania genus assay and the multiplexed positive IC assay were evaluated with known PCR inhibitors. The inhibitors tested were: hemoglobin, hemin, ferric ammonium citrate (FAC), and EDTA. Presence of inhibition was defined as a false negative result (negative result for a true positive sample). The inhibitor concentrations tested and the maximum inhibitor concentration that did not induce inhibition in the Leishmania genus and positive IC assays are listed in the table below.
| PCR Inhibitor | Concentrations Tested(final concentration in PCR reaction tube) | Maximum Concentration without Inhibition Detected | |
|---|---|---|---|
| Leishmania Genus Assay | IC Assay | ||
| Hemoglobin | 0.03, 0.06, 0.12, 0.24, 0.48, 0.96,and 1.92 µg/µl | 1.92 µg/µl | 0.96 µg/µl |
| Hemin | 0.03, 0.06, 0.12, 0.24, 0.48, 0.96,and 1.92 µg/µl | 0.96 µg/µl | 0.96 µg/µl |
| FAC | 0.03125, 0.0625, 0.125, 0.25, 0.50,and 1.00% | 0.0625% | 0.0625% |
| EDTA | 0.3125, 0.625, 1.25, 2.5, 5, and 10mM | 1.25 mM | 1.25 mM |
Table 10. Results of Testing the Effects of Known Inhibitors on the Leishmania Genus and IC Assays
Conclusion from Nonclinical and Clinical Performance Testing
The conclusions drawn from the nonclinical and clinical testing, and comparative methodology review, demonstrate the effectiveness of the SMART Leish for the intended use.
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Image /page/10/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle or bird symbol on the right side. On the left side, the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the bird symbol.
Food and Drug Administration 0903 New Hampshire Avenue Silver Spring, MD 20993
Department of the Army U.S Army Medical Material Development Activity c/o Robert E. Miller, Ph.D., RAC Director, Division of Regulated Activities and Compliance 1430 Veterans Drive Fort Detrick, MD 21702-5009
Re: K081868 Trade/Device Name: SMART Leish PCR Assay Regulation Number: 21 CFR 866.3870 Regulation Name: Trypanosoma spp. Serological Reagents Regulatory Class: Class I Product Code: OUZ Dated: May 23, 2011 Received: May 24, 2011
MAY 2 5 2011
Dear Dr. Miller:
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. 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.
If your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter will allow you to begin marketing your device as described in your Section 510(k) premarket
{11}------------------------------------------------
Page 2 -
notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. 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
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.
Sincerely yours,
Say A. Haps
Sally A. Hojvat, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
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INDICATIONS FOR USE STATEMENT
510(k) Number (if known):
Device Name: SMART Leish .
Indications for Use:
The SMART Leish is a qualitative diagnostic real-time PCR test for the rapid detection of Leishmania species and the identification of L. major in skin lesion scrapings and punch biopsies from individuals suspected of having cutaneous leishmaniasis. The test utilizes real-time polymerase chain reaction assay on the Cepheid SmartCycler® II Dx to detect Leishmania species and L. major. The SMART Leish is indicated for use in patients with clinical presentations and travel history.
WARNING
The SMART Leish is intended for use only by trained laboratory personnel in Department of Defense laboratories. Clinical performance has not been established with strains other than L. major.
Prescription Use (PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
Freddee L. Poole
Division Sign-Off
vision Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
210(k)_14.08186 8 SMART Leish Page 1 of 1 510(k) Notification
§ 866.3870
Trypanosoma spp. serological reagents.(a)
Identification. Trypanosoma spp. serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies toTrypanosoma spp. in serum. The identification aids in the diagnosis of trypanosomiasis, a disease caused by parasitic protozoans belonging to the genusTrypanosoma. Trypanosomiasis in adults is a chronic disease characterized by fever, chills, headache, and vomiting. Central nervous system involvement produces typical sleeping sickness syndrome: physical exhaustion, inability to eat, tissue wasting, and eventual death. Chagas disease, an acute form of trypanosomiasis in children, most seriously affects the central nervous system and heart muscle.(b)
Classification. Class I (general controls).