ESENSOR CF GENOTYPING TESTM ESENSOR SYSTEM, ESENSOR CYSTIC FIBROSIS GENOTYPING TEST, ESENSOR INSTRUMENT, MODELS XT-8

• K Number: K090901
• Device Name: ESENSOR CF GENOTYPING TESTM ESENSOR SYSTEM, ESENSOR CYSTIC FIBROSIS GENOTYPING TEST, ESENSOR INSTRUMENT, MODELS XT-8
• Manufacturer: OSMETECH MOLECULAR DIAGNOSTICS
• Date Cleared: 2009-07-06 ( 96 days)
• Product Code: NUA NSU
• Regulation Number: 866.5900

Intended Use

The eSensor® CF Genotyping Test is an in vitro diagnostic device used to simultaneously detect and identify a panel of mutations and variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene in genomic DNA samples isolated from human peripheral whole blood specimens. The panel includes mutations and variants recommended by the 2004 American College of Medical Genetics (ACMG). The eSensor® CF Genotyping Test is a qualitative genotyping test that provides information intended to be used for cystic fibrosis carrier screening as recommended by ACMG and the 2005 American College of Obstetricians and Gynecologists (ACOG) for adults of reproductive age, as an aid in newborn screening for cystic fibrosis, and in confirmatory diagnostic testing for cystic fibrosis in newborns and children. The test is not indicated for use in fetal diagnostic or pre-implantation testing. This test is also not indicated for stand-alone diagnostic purposes and results should be used in conjunction with other available laboratory and clinical information. The eSensor® CF Genotyping Test is intended for use on the eSensor® XT-8 System.

Device Description

The eSensor® CF Genotyping Test on the eSensor® XT-8 System is an in vitro diagnostic device for performing hybridization and genotyping of multiple mutations and/or polymorphisms in an amplified DNA sample. A single-use, disposable test cartridge is used to perform hybridization and genotyping. The cartridge contains an EEPROM chip which transmits the cartridge lot number, expiration date and protocol identity to the XT-8 instrument. The analysis process for each sample consists of three steps: 1) Genomic DNA isolated from whole blood obtained using EDTA as anti-coagulant is combined with PCR Mix and Taq polymerase enzyme and is subjected to amplification of target sequences by PCR using a thermal cycler. 2) Amplified DNA is treated with exomuclease enzyme to generate single-stranded target DNA. 3) Single-stranded, amplified target DNA is mixed with hybridization and genotyping reagents and transferred to an eSensor® CF Genotyping Tet cartinge, and the cartridge is inserted in the eSensor® XT-8 Instrument controls the circulation of the sample inside the cartridge to allow hybridization at a controlled temperature and then detects and genotypes the sample by voltammetry. Genotyping of the test panel polymorphisms is achieved by a sandwich assay principle: 1) Each pair of electrodes contains a different synthetic oligonucleotide capture probe which is complementary to one of the target DNA fragments. 2) The hybridization reagents contain pairs of ferrocenc-labeled synthetic oligonucleotide signal probes; one member of each pair is complementary to the major allele sequence of the target polymorphism, while the second member of the pair is complementary to the minor allele sequence. Each member of the probe pair has a ferrocene label with a different oxidation potential for each allele. 3) Single-stranded, amplificd target DNA hybridizes to its specific capture probe, and in turn hybridizes to the allele-specific, ferrocenc-labeled signal probe. 4) Each electrode of the array is analyzed by voltammetry; the target polymorphism is determined by the location of the electrode containing the capture probe, and the genotype is identified by the ratio of signals from the allclespecific ferrocene labels. The array also includes positive and negative confirm the hybridization reaction and detect non-specific signals. Upon completion of the test, the EEPROM chip on the cartridge contains information that prevents its re-use with a new sample. The eSensor® XT-8 instrument analyzes the results and provides a report of the test results.

Predicate Devices

K060543, K051435, K063787, K073720

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a chemical and electrical process for genotyping based on hybridization and voltammetry. There is no mention of algorithms, learning, or adaptive processing typically associated with AI/ML. The analysis is based on the ratio of signals from allele-specific labels, which is a direct measurement, not an interpretation or prediction derived from AI/ML.

Therapeutic

No
This device is an in vitro diagnostic (IVD) device used for genotyping the CFTR gene to aid in cystic fibrosis carrier screening and diagnosis. It provides information for diagnosis rather than directly treating or preventing a disease.

Diagnostic

Yes

The device is explicitly stated as an "in vitro diagnostic device" in multiple sections, including "Intended Use / Indications for Use" and "Device Description". It is used for "confirmatory diagnostic testing for cystic fibrosis in newborns and children."

SaMD (Software as a Medical Device)

No

The device description explicitly details hardware components like a disposable test cartridge with an EEPROM chip and the eSensor® XT-8 Instrument, which controls the process and performs detection. This is not a software-only device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

The document explicitly states in the "Intended Use / Indications for Use" section: "The eSensor® CF Genotyping Test is an in vitro diagnostic device..."

Furthermore, the "Device Description" section also refers to it as "an in vitro diagnostic device".

The intended use of the device is to detect and identify mutations in the CFTR gene in genomic DNA samples isolated from human peripheral whole blood specimens, which is a classic definition of an in vitro diagnostic test.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The eSensor® CF Genotyping Test is an in vitro diagnostic device used to simultaneously detect and identify a panel of mutations and variants in the cystic fibrosis transmembrance regulator (CFTR) gene in genomic DNA samples isolated from human peripheral whole blood specimens. The panel includes mutations and variants recommended by the 2004 American College of Medical Genetics (ACMG). The eSensor® CF Genotyping Test is a qualitative genotyping test that provides information intended to be used for cystic fibrosis carrier screening as recommended by ACMG and the 2005 American College of Obstetricians and Gynecologists (ACOG) for adults of reproductive age, as an aid in newborn screening for cystic fibrosis, and in confirmatory diagnostic testing for cystic fibrosis in newborns and children. The test is not in fittal diagnostic or preimplantation testing. This test is also not indicated for stand-alone diagnostic purposes and results should be used in conjunction with other available laboratory and clinical information.

Product codes (comma separated list FDA assigned to the subject device)

NUA, NSU

Device Description

The eSensor® CF Genotyping Test on the eSensor® XT-8 System is an in vitro diagnostic device for performing hybridization and genotyping of multiple mutations and/or polymorphisms in an amplified DNA sample. A single-use, disposable test cartridge is used to perform hybridization and genotyping. The cartridge contains an EEPROM chip which transmits the cartridge lot number, expiration date and protocol identity to the XT-8 instrument.

The analysis process for each sample consists of three steps: 1) Genomic DNA isolated from whole blood obtained using EDTA as anti-coagulant is combined with PCR Mix and Taq polymerase enzyme and is subjected to amplification of target sequences by PCR using a thermal cycler. 2) Amplified DNA is treated with exomuclease enzyme to generate single-stranded target DNA. 3) Single-stranded, amplified target DNA is mixed with hybridization and genotyping reagents and transferred to an eSensor® CF Genotyping Tet cartinge, and the cartridge is inserted in the eSensor® XT-8 Instrument controls the circulation of the sample inside the cartridge to allow hybridization at a controlled temperature and then detects and genotypes the sample by voltammetry

Genotyping of the test panel polymorphisms is achieved by a sandwich assay principle: 1) Each pair of electrodes contains a different synthetic oligonucleotide capture probe which is complementary to one of the target DNA fragments. 2) The hybridization reagents contain pairs of ferrocenc-labeled synthetic oligonucleotide signal probes; one member of each pair is complementary to the major allele sequence of the target polymorphism, while the second member of the pair is complementary to the minor allele sequence. Each member of the probe pair has a ferrocene label with a different oxidation potential for each allele. 3) Single-stranded, amplificd target DNA hybridizes to its specific capture probe, and in turn hybridizes to the allele-specific, ferrocenc-labeled signal probe. 4) Each electrode of the array is analyzed by voltammetry; the target polymorphism is determined by the location of the electrode containing the capture probe, and the genotype is identified by the ratio of signals from the allclespecific ferrocene labels. The array also includes positive and negative confirm the hybridization reaction and detect non-specific signals.

Upon completion of the test, the EEPROM chip on the cartridge contains information that prevents its re-use with a new sample. The eSensor® XT-8 instrument analyzes the results and provides a report of the test results.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Adults of reproductive age, newborns and children.

Intended User / Care Setting

Prescription Use Only

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

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Site to Site, Operator to Operator, Day to Day, Run to Run and sample reproducibility
A reproducibility study was performed over 5 non consecutive days at 3 different sites and 1 internal site. Each site performed the testing twice each day, using two different operators and the same testing materials. Twenty two (22) gDNA samples containing positive calls for all ACOG/ACMG panel mutations in addition to the 5/7/9T polymorphism were used. For practical considerations, the 22 samples were divided into 2 set of 11 samples, so that the sample set was run in duplicate to evaluated intra assay reproducibility using a single kit. Only one lot of materials was used for this study.
The study showed 100% agreement in final correct calls across all sites and operators (Total of 1320 samples).

Lot to Lot Reproducibility
A total of 21 genomic DNA samples covering all possible genotypes were tested using three different kit lots and tested using the eSensor® CF Genotyping Test. The data were evaluated after first-pass results and following additional runs for no-calls. All samples gave 100% correct calls when compared with DNA sequencing. No impact of kit lot observed in this study.

Genomic DNA Extraction Reproducibility
A total of 20 whole blood samples of different genotypes were extracted by three commonly used extraction methods and tested using the eSensor® CF Genotyping Test. The data were evaluated after first-pass results and following additional runs for no-calls. All samples gave 100 correct calls when compared with DNA sequencing. There was no impact of extraction method observed in this study.

Method Comparison
In a method comparison study, a total of 112 gDNA samples extracted from whole blood with A260-280 ratios of 1.2-2.5 were genotyped using the eSensor® CF Genotyping Test and DNA sequencing. All samples gave 100% agreement with DNA sequencing.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Reproducibility: 100% Final Correct calls agreement across sites and operators.
Lot to Lot Reproducibility: 100% correct calls after additional runs for no-calls.
Genomic DNA Extraction Reproducibility: 100% correct calls after additional runs for no-calls.
Method Comparison: 100% agreement with DNA sequencing for all samples.

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.

K060543, K051435, K063787, K073720

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

Regulation Number and Section

§ 866.5900 Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation detection system.

(a)
Identification. The CFTR gene mutation detection system is a device used to simultaneously detect and identify a panel of mutations and variants in the CFTR gene. It is intended as an aid in confirmatory diagnostic testing of individuals with suspected cystic fibrosis (CF), carrier identification, and newborn screening. This device is not intended for stand-alone diagnostic purposes, prenatal diagnostic, pre-implantation, or population screening.(b)
Classification. Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: CFTR Gene Mutation Detection System.” See § 866.1(e) for the availability of this guidance document.

Summary

0

KU90901

510(k) SUMMARY: eSensor® CF Genotyping Test on XT-8 System

Preparation Date: June 12, 2009

Submitted By:

Osmetech Molecular Diagnostics 757 South Raymond Avenue Pasadena, CA 91105 USA Fax: [phone] Phone: [phone]

JUL - 6 2009

Contacts:

Robert Dicheck, Vice President - Quality & Regulatory Affairs (Official Correspondent) Aviva Jacobs, Ph.D., Associate Director - Product Development (Project Manager)

Proprietary Names and Classifications:

For the assav: eSensor® CF Genotyping Test (Kit) Regulation: 21CFR 866.5900 Panel: Immunology (82) Classification: II Product Code: NUA - Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Gene Mutation Detection System

For the instrument: eSensor® XT-8 Instrument (System) Regulation: 21CFR 862.2570 Panel: Clinical Chemistry (75) Classification: II Product Code: NSU - Instrument for Clinical Multiplex Test Systems

Common name:

Cystic Fibrosis Transmembrance Regulator (CFTR) Gene Mutation Detection System

Intended uses:

The eSensor® CF Genotyping Test is an in vitro diagnostic device used to simultaneously detect and identify a panel of mutations and variants in the cystic fibrosis transmembrance regulator (CFTR) gene in genomic DNA samples isolated from human peripheral whole blood specimens. The panel includes mutations and variants recommended by the 2004 American College of Medical Genetics (ACMG). The eSensor® CF Genotyping Test is a qualitative genotyping test that provides information intended to be used for cystic fibrosis carrier screening as recommended by ACMG and the 2005 American College of Obstetricians and Gynecologists (ACOG) for adults of reproductive age, as an aid in newborn screening for cystic fibrosis, and in confirmatory diagnostic testing for cystic fibrosis in newborns and children. The test is not in fittal diagnostic or preimplantation testing. This test is also not indicated for stand-alone diagnostic purposes and results should be used in conjunction with other available laboratory and clinical information.

Special conditions for use statement(s):

For Prescription Use Only

The cScasor® CF Genotyping Test is an in vitro diagnostic device intended for genotyping multiple mutations or polymorphisms in an amplified DNA sample utilizing electrochemical detection technology, for use on the eSensor® XT-8 Instrument.

1

Predicate devices:

eSensor® CFCD System, K060543 and K051435 InPlex™ CF Molecular Test, K063787 eSensor® XT-8 Instrument, K073720

Device Description:

The eSensor® CF Genotyping Test on the eSensor® XT-8 System is an in vitro diagnostic device for performing hybridization and genotyping of multiple mutations and/or polymorphisms in an amplified DNA sample. A singleuse, disposable test cartridge is used to perform hybridization and genotyping. The cartridge contains an EEPROM chip which transmits the cartridge lot number, expiration date and protocol identity to the XT-8 instrument.

The analysis process for each sample consists of three steps: 1) Genomic DNA isolated from whole blood obtained using EDTA as anti-coagulant is combined with PCR Mix and Taq polymerase enzyme and is subjected to amplification of target sequences by PCR using a thermal cycler. 2) Amplified DNA is treated with exomuclease enzyme to generate single-stranded target DNA. 3) Single-stranded, amplified target DNA is mixed with hybridization and genotyping reagents and transferred to an eSensor® CF Genotyping Tet cartinge, and the cartridge is inserted in the eSensor® XT-8 Instrument controls the circulation of the sample inside the cartridge to allow hybridization at a controlled temperature and then detects and genotypes the sample by voltammetry

Genotyping of the test panel polymorphisms is achieved by a sandwich assay principle: 1) Each pair of electrodes contains a different synthetic oligonucleotide capture probe which is complementary to one of the target DNA fragments. 2) The hybridization reagents contain pairs of ferrocenc-labeled synthetic oligonucleotide signal probes; one member of each pair is complementary to the major allele sequence of the target polymorphism, while the second member of the pair is complementary to the minor allele sequence. Each member of the probe pair has a ferrocene label with a different oxidation potential for each allele. 3) Single-stranded, amplificd target DNA hybridizes to its specific capture probe, and in turn hybridizes to the allele-specific, ferrocenc-labeled signal probe. 4) Each electrode of the array is analyzed by voltammetry; the target polymorphism is determined by the location of the electrode containing the capture probe, and the genotype is identified by the ratio of signals from the allclespecific ferrocene labels. The array also includes positive and negative confirm the hybridization reaction and detect non-specific signals.

Upon completion of the test, the EEPROM chip on the cartridge contains information that prevents its re-use with a new sample. The eSensor® XT-8 instrument analyzes the results and provides a report of the test results.

Comparison to technological features of the predicate devices: The following is a comparison of the Osmetech Molecular Diagnostics eSensor® CF Genotyping Test on the XT-8 System to the predicates.

| Characteristic | eSensor® CFCD System
(Predicate 1: K060543 and K051435) | InPlex™ CF Molecular Test (Predicate
2: K063787) | eSensor® CF Genotyping Test |
|---------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------|
| Test type | Qualitative genetic test for single nucleotide
polymorphism detection | Qualitative genetic test for single
nucleotide polymorphism detection | Same as predicates 1 and 2 |
| Sample Type | Genomic DNA obtained from a human whole
blood sample | Genomic DNA obtained from a human
whole blood sample | Same as predicates 1 and 2 |
| Target of detection | Single-nucleotide polymorphism | Single-nucleotide polymorphism | Same as predicates 1 and 2 |
| DNA extraction | Performed off-line | Performed off-line | Same as predicates 1 and 2 |
| Genes | CFTR | CFTR | Same as predicates 1 and 2 |
| Number of Loci genotyped | 23 mutations and 1 variant as recommended by
ACMG (2004)/ACOG (2005). | 23 mutations and 4 variants
recommended by ACMG
(2004)/ACOG (2005). | Same as predicates 1 |
| Genotyping reaction
location | Test cartridge | Microfluidic card | Same as predicate 1 |
| Genotyping principle | Sandwich hybridization test | Fluorometric resonance
energy transfer (FRET) | Same as predicate 1 |
| Instrument operating system | eSensor® Instrument Model 4800 | Multi-well fluorometer | eSensor® Instrument Model XT-8
Random access compatible with multiple
simultaneous test types. |
| Assay results | Assay signal results are interpreted by a
software program and are assigned result that
is presented to the end-user in a report format | InPlex™ CF Molecular Test
Call Reporting Software | Assay signal results are interpreted by a
software program and are assigned a
result that is presented to the end-user in
a report format |

2

Performance Characteristics:

Site to Site, Operator to Operator, Day to Day, Run to Run and sample reproducibility

A reproducibility study was performed over 5 non consecutive days at 3 different sites and 1 internal site). Each site performed the testing twice each day, using two different operators and the same testing materials. Twenty two (22) gDNA samples containing positive calls for all ACOG/ACMG panel mutations in addition to the 5/7/9T polymorphism were used. For practical considerations, the 22 samples were divided into 2 set of 11 samples, so that the sample set was run in duplicate to evaluated intra assay reproducibility using a single kit. Only one lot of materials was used for this study.

| By Site | By
Operator | Number of
Sample
Replicates | First-
pass
Correct
calls | First-pass
No-calls | First-
pass
Miscalls | Final
Correct
calls | Final
incorrect
calls | %
Agreement |
|-----------------------|----------------|-----------------------------------|------------------------------------|------------------------|----------------------------|---------------------------|-----------------------------|----------------|
| Site A
(Internal) | Operator 1 | 220 | 208 | 12 | 0 | 220 | 0 | 100% |
| | Operator 2 | 220 | 216 | 4 | 0 | 220 | 0 | 100% |
| Site B:
(External) | Operator 1 | 220 | 215 | 5 | 0 | 220 | 0 | 100% |
| | Operator 2 | 220 | 211 | 9 | 0 | 220 | 0 | 100% |
| Site C
(External) | Operator 1 | 220 | 216 | 4 | 0 | 220 | 0 | 100% |
| | Operator 2 | 220 | 218 | 2 | 0 | 220 | 0 | 100% |
| | Total | 1320 | 1284 | 36 | 0 | 1320 | 0 | 100% |

Summary of Inter-laboratory, Inter-operator, Reproducibility Results

Summary of Reproducibility Results by Sample Genotype and Site

| Sample Genotype by
Sequencing | 5/7/9T | Number of Sample Replicates
tested by
eSensor® CF
Genotyping Test | Site A
Site A | Site B
Site B | Site C
Site C | Number of CF Sample Calls Before Repeat Testing | Site A
Correct
Calls | Site B
Correct
Calls | Site C
Correct
Calls | No
Calls | Miscalls | Number of CF Sample Calls After Repeat Testing | Site A
Correct
calls | Site B
Correct
Calls | Site C
Correct
calls | No Calls |
|----------------------------------|--------|----------------------------------------------------------------------------|------------------|------------------|------------------|-------------------------------------------------|----------------------------|----------------------------|----------------------------|-------------|----------|------------------------------------------------|----------------------------|----------------------------|----------------------------|----------|
| 1717-1G>A | 7T/7T | 60 | 60 | 60 | 59 | 59 | 59 | 3 | 0 | 60 | 60 | 60 | 0 | | | |
| 1898+1G>A/ΔF508 | 7T/9T | 60 | 60 | 60 | 59 | 60 | 58 | 3 | 0 | 60 | 60 | 60 | 0 | | | |
| 2184delA/ΔF508 | 7T/9T | 60 | 60 | 60 | 58 | 60 | 58 | 4 | 0 | 60 | 60 | 60 | 0 | | | |
| 3120+1G>A/621+1G>T | 7T/9T | 60 | 60 | 60 | 59 | 60 | 59 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| 2789+5G>A/2789+5G>
A | 7T/7T | 60 | 60 | 60 | 59 | 60 | 59 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| 3659delC/ΔF508 | 7T/9T | 60 | 60 | 60 | 59 | 59 | 60 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| 3849+10KbC>T/ΔF508 | 7T/9T | 60 | 60 | 60 | 60 | 59 | 58 | 3 | 0 | 60 | 60 | 60 | 0 | | | |
| 621+1G>T/G85E | 7T/9T | 60 | 60 | 60 | 58 | 60 | 60 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| 711+1G>T/621+1G>T | 7T/9T | 60 | 60 | 60 | 59 | 60 | 60 | 1 | 0 | 60 | 60 | 60 | 0 | | | |
| A455E/621+1G>T | 9T/9T | 60 | 60 | 60 | 59 | 60 | 59 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| Δ1507 | 7T/7T | 60 | 60 | 60 | 60 | 60 | 60 | 0 | 0 | 60 | 60 | 60 | 0 | | | |
| G542X | 7T/9T | 60 | 60 | 60 | 59 | 59 | 60 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| G551D/R347P | 7T/7T | 60 | 60 | 60 | 59 | 59 | 60 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| N1303K | 7T/9T | 60 | 60 | 60 | 60 | 60 | 60 | 0 | 0 | 60 | 60 | 60 | 0 | | | |
| R1162X | 7T/7T | 60 | 60 | 60 | 60 | 60 | 59 | 1 | 0 | 60 | 60 | 60 | 0 | | | |
| R117H/ΔF508 | 5T/9T | 60 | 60 | 60 | 60 | 60 | 59 | 1 | 0 | 60 | 60 | 60 | 0 | | | |
| R334W | 7T/7T | 60 | 60 | 60 | 60 | 59 | 59 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| R553X/G551D | 7T/7T | 60 | 60 | 60 | 59 | 60 | 60 | 1 | 0 | 60 | 60 | 60 | 0 | | | |
| R560T/ΔF508 | 7T/9T | 60 | 60 | 60 | 59 | 60 | 60 | 1 | 0 | 60 | 60 | 60 | 0 | | | |
| W1282X | 5T/7T | 60 | 60 | 60 | 59 | 60 | 59 | 2 | 0 | 60 | 60 | 60 | 0 | | | |
| WT | 7T/7T | 60 | 60 | 60 | 60 | 60 | 60 | 0 | 0 | 60 | 60 | 60 | 0 | | | |
| R117H/ΔF508 | 5T/9T | 60 | 60 | 60 | 60 | 60 | 60 | 0 | 0 | 60 | 60 | 60 | 0 | | | |

3

Lot to Lot Reproducibility

A total of 21 genomic DNA samples covering all possible genotypes were tested using three different kit lots and tested using the eSensor® CF Genotyping Test. The data were evaluated after first-pass results and following additional runs for no-calls. All samples gave 100% correct calls when compared with DNA sequencing. No impact of kit lot observed in this study. The following table summarizes the results of lot to lot reproducibility study.

| LOT | Samples
Tested | First pass
correct
calls | First pass | | Final | | Final
correct
calls | %
Agreement |
|-------|-------------------|--------------------------------|------------|----------|----------|----------|---------------------------|----------------|
| | | | No Calls | Miscalls | No Calls | Miscalls | | |
| 1 | 21 | 21 | 0 | 0 | 0 | 0 | 21 | 100% |
| 2 | 21 | 21 | 0 | 0 | 0 | 0 | 21 | 100% |
| 3 | 21 | 20 | 1 | 0 | 0 | 0 | 21 | 100% |
| Total | 63 | 62 | 1 | 0 | 0 | 0 | 63 | 100% |

Genomic DNA Extraction Reproducibility

A total of 20 whole blood samples of different genotypes were extracted by three commonly used extraction methods and tested using the eSensor® CF Genotyping Test.. The data were evaluated after first-pass results and following additional runs for no-calls. All samples gave 100 correct calls when compared with DNA sequencing. There was no impact of extraction method observed in this study. The following table summarizes the results of extraction reproducibility study.

Extraction Method	# Samples Tested/Genotype	First pass correct calls	First pass No Calls	Miscalls	Final correct calls	Final Agreement
A	20	17	3	0	20	100
B	20	18	2	0	20	100
C	20	*19	1	0	20	100

4

Method Comparison

In a method comparison study, a total of 112 gDNA samples extracted from whole blood with A260-280 ratios of 1.2-2.5 were genotyped using the eSensor® CF Genotyping Test and DNA sequencing.

All samples gave 100% agreement with DNA sequencing. The following table summarizes the results of the method comparison study.

| | | | Sequencing
Calls | 1st Pass CF GT Calls | | | % Agreement | | FINAL CF GT Calls | | | % Agreement | | | |
|---------------------------------|-----------------------|-----|---------------------|----------------------|----------|-------------|-----------------------------------------------------------|-------|-------------------|-----|-----------|-------------|---------|--------|-------|
| | | | | | | | | | | | | | | | |
| Genotype by
sequencing | Calls per
mutation | Pos | Neg | Pos | Neg | No
Calls | Overall | Pos | Neg | Pos | Neg | No
Calls | Overall | Pos | Neg |
| AF508 | 112 | 47 | ર્ણ ર | 46 | 65 | | 99.1 | 97.9 | 100 | 47 | 65 | 0 | 100 | 100 | 1 (AQ |
| G542X | 112 | 7 | ! OS | 6 | 1 05 | | ਰੇਰੇ I | 85.7 | 100 | 7 | 105 | 0 | 1 00 | 100 | 1 00 |
| WI 282X | 112 | б | 100 | 6 | 1 06 | 0 | 100 | 1 000 | 100 | 6 | 106 | 0 | 1 00 | 100 | I (X) |
| GSSID | 112 | 8 | 104 | 8 | 104 | 0 | 100 | 100 | 100 | ,8 | 104 | 0 | 1 00 | 100 | 100 |
| 621+1G>T | 112 | 7 | રે OS | 7 | 1 05 | 0 | 100 | 100 | 100 | 7 | 105 | D | 1 00 | 100 | 1 (ਮ) |
| N1303K | 112 | 8 | 1 04 | 7 | 104 | | ਉਹ | 87.5 | ﺍ 00 | 8 | 104 | 0 | 100 | 100 | 100 |
| RSSX | 112 | 4 | 108 | 4 | 108 | 0 | 1 (A0 | 100 | 1 00 | 4 | 108 | 0 | 1 00 | 100 | 1 00 |
| AI207 | 112 | 3 | I 09 | 3 | I tra | 0 | 1 (X) | 1 00 | 100 | 3 | 109 | 0 | 100 | 100 | 1 00 |
| 3120+1G>A | 112 | د | 110 | 2 | I I Q | 0 | 100 | 100 | 1 00 | 2 | 110 | 0 | 100 | 100 | 1 00 |
| 3849+10kbC>T | 112 | 5 | 107 | 5 | 107 | 0 | 100 | 100 | 100 | 5 | 107 | 0 | ! (10 | I ()() | 100 |
| RITTH | 112 | 8 | 104 | 8 | 104 | 0 | 100 | 1 00 | 100 | 8 | 104 | 0 | 100 | 1 00 | ﺍ |
| 1717-1G>A | 112 | 5 | 107 | 5 | 107 | 0 | 1 00 | 100 | 100 | 5 | 107 | 0 | 1 00 | 100 | 100 |
| 2789+5G>A | 112 | 5 | 1 07 | 5 | 107 | 0 | 1 00 | 100 | ા રીવ | 5 | 107 | 0 | 100 | 100 | 100 |
| R334W | 112 | 4 | 1 08 | 4 | ા 08 | 0 | 100 | 100 | ﺍ 00 | 4 | 1 08 | 0 | 100 | 100 | 100 |
| K347P | 112 | 4 | 1 08 | 4 | 108 | 0 | 100 | 100 | ﺍ 00 | 4 | 108 | 0 | 100 | 00 | 100 |
| 711+1G>T | 112 | 4 | 1 68 | 4 | 108 | 0 | 100 | 100 | 1 00 | 4 | 1 08 | 0 | 100 | 1 00 | 100 |
| R 560 T | 112 | 4 | 1 08 | ﻟﺪ | 108 | - | ਰੇਰੇ | 75.0 | 100 | 4 | 1 08 | 0 | 100 | 100 | 100 |
| RITE2X | 112 | 8 | 104 | 8 | 104 | 0 | 100 | 100 | 100 | 8 | 104 | 0 | 100 | 100 | 100 |
| 3659dclC | 112 | 4 | 108 | 4 | 108 | 0 | 100 | 100 | 100 | 4 | 108 | 0 | 100 | 100 | 100 |
| A455E | 112 | 2 | 110 | 2 | 110 | 0 | 100 | 1 60 | 100 | 2 | 110 | 0 | 100 | 100 | 1 00 |
| C82E | II2 | 7 | 105 | 7 | દ્દ OS | 0 | 100 | 100 | 100 | 7 | 105 | 0 | 100 | 100 | 1 00 |
| 2184dc1A | 112 | 2 | 110 | 2 | 110 | 0 | 100 | 100 | 100 | 2 | 110 | 0 | 100 | 100 | 1 00 |
| 1898+1G>V | 112 | 2 | 110 | 2 | 110 | 0 | 100 | 100 | 100 | 2 | 110 | 0 | 100 | 100 | 1 00 |
| | | | | | | | Reflex Polymorphism | | | | | | | | |
| IVS8--8 5T/7T/9T
Variant (S) | 112 | 6 I | 106 | 6 1 | 1196 | 0 | 1 00 | 1 00 | 100 | 6 | 11)S | 0 | 100 | 100 | 1 00 |
| | | | | | | | Polymorphisms not specifically genotyped | | | | | | | | |
| 1507V | 112 | - | 11 | 1 | 111 | 0 | 100 | 100 | 100 | 1 | 111 | 0 | 100 | 1 00 | 1 00 |
| FS08C | 112 | 1 | l l | | 111 | 0 | ﺍ 00 | 100 | ા ೧೧ | l | 1 I | 0 | 100 | 100 | 100 |
| | | | | | | | Potentially interfering mutations not part of assay panel | | | | | | | | |
| 2183AA>G | 112 | 2 | 110 | 2 | 110 | 0 | 100 | 100 | 100 | 2 | l 10 | 0 | 1 00 | 1 00 | 100 |
| RITL | 112 | 】 | l 11 | l | 111 | 0 | 100 | 100 | 100 | ﺎ | l ] I
| 0 | 100 | 1 00 | 100 |
| Grand Total | 2688 | 162 | 2526 | ા રેક્ષે | 2526 | 4 | 99,9 | 97.5 | 100 | 162 | 2526
3 | 0 | 100 | 100 | 100 |

s For the purpose of the IVSS-577779T Variant, "Positive" samples are east one opy of the ST allele while "Negative" samples are regarded as having only the 7T and/or 9T allele. $1 sample is 5T/5T Mutant.

The number of positive sequencing calls is greater than the number of inclusion of compond heleruzyeus sumples.

The Grand Total consists of the total number of sells for mutations, the 1 57/57 sample included, polymophisms not specifically genoryed, and the potentially interfeing not part of assay panel. 1507V, F508C, 2183AA>G, and R117L are non-panel polymorphism containing samples correctly called as Wild-Type, which are not included in the grand total for calls. No samples with 1506V were tested.

5

Other Characteristics of the eSensor® CF Genotyping Test:

Characteristic	Result
Limit of detection	Two genomic DNA samples of different genotypes were extracted from whole blood stored in EDTA and serially diluted and
tested 20 times each at input amounts of 0.01, 0.1, 1, 10, 100, 500 and 1000 ng/PCR. using the eSensor® CF Genotyping Test.
An additional run was performed for tests that gave a first pass no call result. All input amounts from 0.01ng to 1000ng for both
samples gave equivalent final performance. (100% Agreement with 98.76%, 95% LCB), (99.95% LCB on a per SNP basis).
The limit of detection was established as the lowest concentration at which no-calls or mis-calls were obtained.

The lower detection limit was determined to be 0.1 ng of purified DNA per reaction and the upper detection limit was
determined to be 1000 ng of purified DNA per reaction. The recommended range of DNA input amounts for the eSensor® CF
Genotyping Test is from 10 to 500 ng. | | | | |
| Interfering substances | Test performance was not affected by addition of the following substances seven whole blood samples of different genotypes
prior to extraction:
Bilurubin (30 mg /dL whole blood). Triglycerides (500 mg/dL whole blood). Hemoglobin (~20g /dL whole blood). EDTA (at a concentration equivalent to 5-fold higher than that provided by a standard EDTA blood collection tube) | | | | |
| Test Limitations | The eSensor® CF Genotyping Test does not identify all possible mutations in the CFTR gene, for example E60X, 1148T,
1078delT, V520F, 2143delT, 3199del6, D1152H, 3876delA, 2183AA>G, R560K, R117L, R347H, G551S, 711+5G>A,
394delTT, and 3905insT. A negative result for all the mutations in this panel does not necessarily indicate that the individual is
negative for cystic fibrosis (carrier or affected status). The mutations included in this test represent >80% of the mutations
carried by Caucasian American adults.

The eSensor® CF Genotyping Test does not identify the 1506V, 1507V, and F508C polymorphisms, thus in the case of AF508
homozygosity, reflex testing by bi-directional DNA sequencing is recommended | | | | |
| Interfering
mutations and polymorphisms | More than 1,000 mutations have been identified in the CFTR gene with varying confidence that they are CF disease causing.
When present in the same region as a panel mutation, they may interfere with genotyping results
The following mutations have been evaluated and demonstrated not to impact the results of the eSensor® CF Genotyping Test
Non-Panel Mutation or Polymorphism ACOG/ACMG Panel Mutation 2183AA>G
R117L 2184delA
R117H | | | | |

6

Kit Stability:

eSensor® CF Genotyping Test kit components should be stored under the appropriate conditions until the expiration date printed on the label:

• · PCR Box containing CF Genotyping Test PCR Mix and Taq Polymerase: Store at -20℃ in a designated pre-PCR area.
• · Cartridges: Store at 10° to 25°C
• · Genotyping Box containing Exonuclease, CF Genotyping Test Signal Buffer, XT-Buffer 2: Store at -20℃ in a designated post-PCR area.

In-process stability has been established for the following components, working reagents and samples:

• · Cartides can be stored for up to 14 days after opening the foil pouches. If stored, cartridges should be kept in their original foil pouch at room temperature in a dry place with the zip-loc closure sealed.
• · Once open, reagents can be stored at -20°C for up to 30 days.
• · Reagents can be thawed up to 3 times.
• Whole blood stored in EDTA can be stored for up to 4 weeks after collection of gDNA for use in the eSensor® CF Genotyping Test.
• · PCR product can be stored at 4°C or -20°C for up to 7 days.
• · Exonuclease-digested PCR product can be stored store at 4℃ or -20℃ for up to 7 days.
• · After combining the exonuclease-digested PCR with hybridization reaction can be loaded on the cartridge and held at ambient temperature for up to 8 hours before initiating hybridization of the cartridge on the XT-8 instrument.

Conclusion:

The above internal and clinical test results support the safety and effectiveness of the eSensor® XT-8 System, and demonstrate substantial equivalence to the predicate device.

eSensor® is a registered trademark of Osmetech and its subsidiaries

7

Image /page/7/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES · USA" around the perimeter. Inside the circle is a stylized symbol featuring three abstract human figures connected by flowing lines, representing the department's focus on health and human well-being.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Food and Drug Administration 10903 New Hampshire Avenue Building 66 Silver Spring, MD 20993

Osmetech Molecular Diagnostics c/o Mr. Robert S. Dicheck, Vice President - Quality & Regulatory Affairs 757 S. Raymond Avenue Pasadena, CA 91105

JUL - 6 2009

Re: K090901

Trade/Device Name: eSensor® CF Genotyping Test Regulation Number: 21 CFR 866.5900 Regulation Name: CFTR (cystic fibrosis transmembrane conductance regulator) gene mutation detection system Regulatory Class: Class II Product Code: NUA, NSU Dated: June 9, 2009 Received: June 10, 2009

Dear Mr. Dicheck:

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. Ilsting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), 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 (OS) regulation (21 CFR Part 820). This letter

8

Page 2 - Mr. Robert S. Dicheck

will allow you to begin marketing your device as described in your Section 510(k) premarket 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 Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR 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 [phone] or [phone] or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Maria mchan

Maria M. Chan, Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

9

Indication(s) for Use

510(k) Number (if known): K090901

Device Name: eSensor® CF Genotyping Test

Intended Use/Indication(s) For Use:

The eSensor® CF Genotyping Test is an in vitro diagnostic device used to simultaneously detect and identify a panel of mutations and variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene in genomic DNA samples isolated from human peripheral whole blood specimens. The panel includes mutations and variants recommended by the 2004 American College of Medical Genetics (ACMG). The eSensor® CF Genotyping Test is a qualitative genotyping test that provides information intended to be used for cystic fibrosis carrier screening as recommended by ACMG and the 2005 American College of Obstetricians and Gynecologists (ACOG) for adults of reproductive age, as an aid in newborn screening for cystic fibrosis, and in confirmatory diagnostic testing for cystic fibrosis in newborns and children. The test is not indicated for use in fetal diagnostic or pre-implantation testing. This test is also not indicated for stand-alone diagnostic purposes and results should be used in conjunction with other available laboratory and clinical information.

The eSensor® CF Genotyping Test is intended for use on the eSensor® XT-8 System.

Prescription Use X. (21 CFR Part 801 Subpart D) And/Or

Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)

Nu Ru

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K090901