(1057 days)
Not Found
No
The description focuses on standard PCR technology, reagent kits, and automated signal interpretation by the instrument, with no mention of AI or ML algorithms for analysis or interpretation.
No.
This device is a qualitative diagnostic test that detects Leishmania species and L. major DNA, which is used to diagnose cutaneous leishmaniasis rather than treat it.
Yes
The "Intended Use / Indications for Use" section explicitly states that "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." This directly identifies its purpose as diagnostic.
No
The device description clearly states that the SMART Leish consists of an assay reagent kit and assay definition files. The kit contains physical reagents in lyophilized bead form. While it includes software components (assay definition files), it is fundamentally a diagnostic test kit with physical reagents and requires a specific hardware instrument (Cepheid SmartCycler® II Dx) for operation.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The device 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." This clearly states its purpose is for diagnosis.
- Sample Type: It uses "skin lesion scrapings and punch biopsies," which are biological specimens taken from the human body.
- Testing Location: The testing is performed "in Department of Defense laboratories," indicating it's done outside of the body (in vitro).
- Method: It utilizes "real-time polymerase chain reaction assay," a common laboratory technique for analyzing biological samples.
- Output: The test provides a "qualitative" result (positive or negative) for the presence of Leishmania species and L. major, which is used to aid in diagnosis.
All of these characteristics align with the definition of an In Vitro Diagnostic device.
No
The document does not contain any mention or indication that this device is authorized under a Predetermined Change Control Plan (PCCP). PCCP authorization would typically be explicitly stated in the clearance letter or within sections discussing modifications or future changes.
Intended Use / Indications for 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.
Product codes (comma separated list FDA assigned to the subject device)
OUZ
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.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
skin lesion scrapings and punch biopsies
Indicated Patient Age Range
adults 18 years or older
Intended User / Care Setting
trained laboratory personnel in Department of Defense laboratories
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
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.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Clinical Performance Testing:
Type: Multi-center, retrospective clinical study
Sample Size: 312 Leishmania genus specimens, 187 Leishmania major specimens.
Key Results for Leishmania genus Assay:
- Clinical Truth Positive: 225
- Clinical Truth Negative: 87
- SMART Leish Positive: 252 (223 true positive, 29 false positive)
- SMART Leish Negative: 60 (2 false negative, 58 true negative)
- 27 of 29 false positives were determined to be Genus positive using sequencing methods.
Key Results for Leishmania major Assay:
- Clinical Truth Positive: 96
- Clinical Truth Negative: 91
- SMART Leish Positive: 100 (92 true positive, 8 false positive)
- SMART Leish Negative: 87 (4 false negative, 83 true negative)
- 3 of 8 false positives were determined to be L. major using sequencing methods.
Analytical Reactivity Study:
Performed with 36 Leishmania strains.
Leishmania genus study: DNA from all 36 Leishmania strains gave robust positive results.
Leishmania major study: 11 of 36 strains were Leishmania major species; all 11 gave robust positive results. None of the 25 non-Leishmania major samples tested positive.
Analytical Specificity Study:
121 DNA samples evaluated.
- Leishmania major assay: 11 L. major strains tested positive. 25 non-Leishmania major samples tested negative. Analytical specificity was 100%. False positive rate 0%, false negative rate 0%.
- Leishmania genus assay: 85 non-target DNA samples were tested; 85 were negative, 2 false positives, and 36 were positive. Analytical specificity was 98.3%, false positive rate 2.4%, false negative rate 0%.
- Two strains of C. fasciculata were low-level cross-reactors for the Leishmania genus assay. No cross-reactors for the L. major assay.
- In silico analysis predicted no cross-reactivity with organisms causing similar clinical presentations.
Analytical Sensitivity (Limit of Detection - LOD) Study:
- Leishmania Genus Assay: Linear Range 0.014-14,260 pg, Assay LOD 0.14 pg (equates to ~4 genome copies with ~200 copies/genome of target gene). Extraction LOD (eLOD) 250 parasites (equates to ~5 genome copies at assay reaction stage).
- L. major Assay: Linear Range 0.2139-21,390 pg, Assay LOD 2.1 pg (equates to 62 genome copies). Extraction LOD (eLOD) 1,000 parasites (equates to ~30 genome copies at assay reaction stage).
Reproducibility Study:
Internal evaluation at three laboratories (WRAIR, BAMC, MAMC) over 5 days.
Leishmania Genus Assay:
- Purified DNA (Low, Medium, High concentrations): Total Agreement 100% for all concentrations. Positive Agreement for Purified DNA: 100%.
- Cultured Parasites (Low, Medium, High concentrations): Total Agreement 100% for all concentrations. Positive Agreement for Cultured Parasites: 100%.
- Mock Human Samples: Negative: Total Agreement 100%. Positive: Total Agreement 97.6% (41/42). Overall Agreement for Mock Human Samples: 98.6% (71/72).
Leishmania major Assay:
- Purified DNA: Low (61% Total, 37/60), Medium (100% Total, 60/60), High (100% Total, 60/60). Positive Agreement for Purified DNA: 87.2% (157/180).
- Cultured Parasites: Low (91.6% Total, 55/60), Medium (100% Total, 60/60), High (100% Total, 60/60). Positive Agreement for Cultured Parasites: 97.2% CI (175/180).
- Mock Human Samples: Negative: Total Agreement 100% (0/0 - no data provided for negative mock human samples, likely because they were not further tested if Leishmania genus assay was negative). Positive: Total Agreement 100% (41/41). Overall Agreement for Mock Human Samples: 100% (41/41).
Interfering Substances Study:
Evaluated known PCR inhibitors: hemoglobin, hemin, ferric ammonium citrate (FAC), and EDTA.
- Hemoglobin: Max concentration without inhibition detected for Leishmania Genus Assay 1.92 µg/µl, IC Assay 0.96 µg/µl.
- Hemin: Max concentration without inhibition detected for Leishmania Genus Assay 0.96 µg/µl, IC Assay 0.96 µg/µl.
- FAC: Max concentration without inhibition detected for Leishmania Genus Assay 0.0625%, IC Assay 0.0625%.
- EDTA: Max concentration without inhibition detected for Leishmania Genus Assay 1.25 mM, IC Assay 1.25 mM.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Key results for Leishmania Genus Assay:
- Positive Percent Agreement (Sensitivity): 223/225 = 99.1%
- Negative Percent Agreement (Specificity): 58/87 = 66.7%
Key results for Leishmania major Assay:
- Positive Percent Agreement (Sensitivity): 92/96 = 95.8%
- Negative Percent Agreement (Specificity): 83/91 = 91.2%
Analytical Specificity:
- Leishmania major assay: 100%
- Leishmania genus assay: 98.3%
False Positive Rate for Analytical Specificity:
- Leishmania major assay: 0%
- Leishmania genus assay: 2.4%
False Negative Rate for Analytical Specificity:
- Leishmania major assay: 0%
- Leishmania genus assay: 0%
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Amizyme-Leishmania spp. Test Kit K842526
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 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).
0
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
1
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.
2
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 controls | |
| for both the extraction process and | ||
| detection assay runs | ||
| Internal - positive control for PCR | ||
| inhibitors in sample | External positive and negative | |
| controls | ||
| Differences | ||
| Instrument Platform | Cepheid SmartCycler | None |
| Automation | Automated DNA amplification cycling, | |
| probe detection, and interpretation of | ||
| results | Manual method | |
| Sample Collection | Skin punch biopsy or scrapings, stored in | |
| ethanol | Serum | |
| Endpoint detection method | PCR amplification of DNA sequences | |
| unique to the organisms of interest with | ||
| detection by hybridization probes | ||
| incorporating reporter dyes | Indirect Immunofluoresence (IFA) | |
| Fluorescent conjugate secondary | ||
| antibodies on a prepared slide |
Table 1. Similarities
3
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 - | ||||
| Leishmania | ||||
| Genus 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.
4
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).
5
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
6
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 of | |||
| target 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 assay |
| reaction stage | ||
| L. major Assay | 1,000 parasites | ~30 genome copies at assay |
| reaction 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.
7
Table 8. Summary of Between Laboratory Reproducibility Results for the Leishmania Genus Assay ﺍﻟﻘﺎﺩﺭ
| Sample
Type | Sample ID | Concentration | Number of
Replicates
(over 5 days) | Laboratory
A
(WRAIR) | Laboratory
B
(BAMC) | Laboratory
C
(MAMC) | Total
Agreemen
t | % Total
Agreemen
t |
|------------------------------------------------------|-----------|----------------------------|------------------------------------------|----------------------------|---------------------------|---------------------------|------------------------|--------------------------|
| | | genome
equivalents/µL | | | | | | |
| Purified
DNA | 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% |
| Positive
Agreement
for Purified
DNA | | | | | | | 180/180 | 100% |
| | | (parasites/
extraction) | | | | | | |
| Cultured
Parasites | 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% |
| Positive
Agreement
for Cultured
Parasites | | | | | | | 180/180 | 100% |
| | | (parasites/
extraction) | | | | | | |
| Mock
Human
Samples | 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% |
| Overall
Agreement
for Mock
Human
Samples | | | | | | | 71/72 | 98.6% |
SMART LEISH 510(k) Summary
:
:
8
Table 9. Summary of between Laboratory Reproducibility Results for the Leishmania major Assay
| Sample
Type | Sample ID | Concentration | Number of
Replicates
(over 5
days) | Laboratory
A
(WRAIR) | Laboratory
B
(BAMC) | Laboratory
C
(MAMC) | Total
Agreem
ent | % Total
Agreement |
|-------------------------------------------------------|-----------|----------------------------|---------------------------------------------|----------------------------|---------------------------|---------------------------|------------------------|----------------------|
| | | genome
equivalents/µL | | | | | | |
| Purified
DNA | 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% |
| Positive
Agreement
for Purified
DNA | | | | | | | 157/180 | 87.2% |
| | | (parasites/
extraction) | | | | | | |
| Cultured
Parasites | 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% |
| Positive
Agreement
for
Cultured
Parasites | | | | | | | 175/180 | 97.2% CI |
| | | (parasites/ | | | | | | |
| | | extraction) | | | | | | |
| Mock
Human
Samples | 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% |
| Overall
Agreement
for Mock
Human
Samples | | | | | | | 41/41 | 100% |
9
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 10
mM | 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
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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