K Number

Device Name

XPERT HEMOSIL FACTOR II & FACTOR V ASSAY

Manufacturer

CEPHEID

Date Cleared

2009-09-18

(417 days)

Product Code

NPQ NPR OOI

Regulation Number

864.7280

Intended Use

The Xpert™ HemosIL® Factor II & Factor V Assay is a qualitative in vitro diagnostic genotyping test for the rapid detection of Factor II and Factor V alleles from sodium citrate or EDTA anticoagulated whole blood. The test is performed on the Cepheid GeneXpert® Dx System. This test is intended to provide rapid results for Factor II (G20210A) and Factor V (Leiden) mutations as an aid in the diagnosis in individuals with suspected thrombophilia.

Device Description

The Cepheid Xpert® HemosIL® Factor II & Factor V Assay is an automated DNA test for detecting Factor II and Factor V normal and mutant alleles directly from sodium citrate or EDTA anticoagulated whole blood specimens. Blood specimens are drawn into either sodium citrate or EDTA anticoagulant tubes. Following brief mixing of the sample, the blood sample and two single-use reagents (Reagent 1 and Reagent 2) that are provided with the assay are transferred to different, uniquely-labeled chambers of the disposable fluidic cartridge (the Xpert HemosIL Factor V cartridge). The user initiates a test from the system user interface and places the cartridge into the GeneXpert® Dx System instrument platform, which performs hands-off real-time, multiplex polymerase chain reaction (PCR) for detection of DNA. In this platform, sample preparation, amplification, and real-time detection are all fully-automated and completely integrated. The GeneXpert Dx System consists of a GeneXpert instrument, personal computer, a barcode scanner and the multi-chambered fluidic cartridges that are designed to complete sample preparation and real-time PCR for detection of Factor II and Factor V normal and mutant alleles in approximately 30 minutes. Each system has 1 to 16 randomly accessible modules that are each capable of performing separate sample preparation and real-time PCR tests. Each module contains a syringe drive for dispensing fluids, an ultrasonic horn for lysing nuclei, and a proprietary I-CORE® thermocycler for performing real-time PCR and detection. The Xpert HemosIL Factor II & Factor V Assay includes reagents for the detection of Factor II and Factor V normal and mutant alleles. The primers and probes in the Xpert HemosIL Factor II & Factor V Assay determine the genotype of the Factor II gene (at position 20210) and the Factor V gene (at position 1691). The test includes a Probe Check Control (PCC) that verifies reagent rehydration, PCR tube filling in the cartridge, probe integrity, and dye stability and an instrument control verifies the instrument is performing properly. Additionally, the sample functions as its own internal control since both normal and mutant gene sequences are detected, and each person tested is expected to have one of these sequence signatures.

More Information

Contact

Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

State

Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

K033612, K033607

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description details a fully automated PCR-based genotyping test with integrated sample preparation and detection. There is no mention of AI or ML being used for analysis or interpretation of results. The performance studies focus on accuracy relative to a gold standard (sequencing) and reproducibility, not on training or validation of an AI/ML model.

Therapeutic

No.
The device is an in vitro diagnostic genotyping test used to detect Factor II and Factor V alleles as an aid in diagnosing thrombophilia; it does not provide treatment or therapy.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the test is "intended to provide rapid results for Factor II (G20210A) and Factor V (Leiden) mutations as an aid in the diagnosis in individuals with suspected thrombophilia." The phrase "aid in the diagnosis" clearly indicates its role as a diagnostic device.

SaMD (Software as a Medical Device)

The device description clearly states it includes hardware components such as the GeneXpert instrument, personal computer, barcode scanner, and disposable fluidic cartridges, in addition to the software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the Xpert™ HemosIL® Factor II & Factor V Assay is a "qualitative in vitro diagnostic genotyping test".
Device Description: The description details how the test is performed on biological samples (whole blood) outside of the body ("in vitro") to provide diagnostic information (detection of Factor II and Factor V alleles).
Purpose: The test is intended to "aid in the diagnosis in individuals with suspected thrombophilia," which is a diagnostic purpose.

The information provided clearly aligns with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

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

NPO, NPR, OOI

Device Description

The GeneXpert Dx System consists of a GeneXpert instrument, personal computer, a barcode scanner and the multi-chambered fluidic cartridges that are designed to complete sample preparation and real-time PCR for detection of Factor II and Factor V normal and mutant alleles in approximately 30 minutes. Each system has 1 to 16 randomly accessible modules that are each capable of performing separate sample preparation and real-time PCR tests. Each module contains a syringe drive for dispensing fluids, an ultrasonic horn for lysing nuclei, and a proprietary I-CORE® thermocycler for performing real-time PCR and detection.

The Xpert HemosIL Factor II & Factor V Assay includes reagents for the detection of Factor II and Factor V normal and mutant alleles. The primers and probes in the Xpert HemosIL Factor II & Factor V Assay determine the genotype of the Factor II gene (at position 20210) and the Factor V gene (at position 1691).

The test includes a Probe Check Control (PCC) that verifies reagent rehydration, PCR tube filling in the cartridge, probe integrity, and dye stability and an instrument control verifies the instrument is performing properly. Additionally, the sample functions as its own internal control since both normal and mutant gene sequences are detected, and each person tested is expected to have one of these sequence signatures.

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

Not Found

Intended User / Care Setting

Operators with no clinical lab experience to experienced clinical laboratory technologists

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

A total of 1018 samples were tested for Factor II by both the Xpert HemosIL Factor II & Factor V Assay, and bi-directional sequencing. A total of 1014 samples were tested for Factor V by both the Xpert HemosIL Factor II & Factor V Assay and bi-directional sequencing. To supplement the homozygous sample size, six human genomic DNA samples homozygous for Factor II and five homozygous for Factor V were also tested by the Xpert HemosIL Factor II & Factor V Assay and bi-directional sequencing. Performance of the Xpert HemosIL Factor II & Factor V Assay was calculated relative to bi-directional sequencing results. The bi-directional sequencing results functioned as the gold standard.

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

Clinical Comparison Study:

Sample Size: 1018 (Factor II), 1014 (Factor V), plus 6 homozygous Factor II samples and 5 homozygous Factor V samples.
Key Results: The Xpert HemosIL Factor II & Factor V Assay demonstrated a 99.3% overall accuracy relative to bi-directional sequencing for both Factor II and Factor V.
Detailed Results:
- Factor II G20210A:
  - Agreement on First Run: WT 95.8% (927/968), HET 96.0% (48/50), HOM 100.0% (7/7). Overall 95.8% (982/1025).
  - Agreement After Repeat Run: WT 99.5% (963/968), HET 96.0% (48/50), HOM 100% (7/7). Overall 99.3% (1018/1025).
- Factor V G1691A:
  - Agreement on First Run: WT 96.1% (860/895), HET 94.7% (108/114), HOM 91.7% (11/12). Overall 95.9% (979/1021).
  - Agreement After Repeat Run: WT 99.3% (889/895), HET 99.1% (113/114), HOM 100.0% (12/12). Overall 99.3% (1014/1021).

Reproducibility Study:

Sample Size: A panel of 5 specimens tested in duplicate by two different operators on 5 different days at each of three sites (3 specimens x 2 times/day x 2 operators per site x 5 days x 3 sites), totaling 300 runs. For between-lot reproducibility, the 5-specimen panel was analyzed two times per day over 5 testing days using each of three assay lots, at a single testing site (5 specimens x 2 runs per day x 3 lots x 5 days), totaling 150 runs.
Key Results:
- By Site (Factor II & Factor V): Overall total agreement by site was 99.7% (299/300).
  - Site 1: 100% (60/60)
  - Site 2: 100% (60/60)
  - Site 3: 98.3% (59/60) (One sample indeterminate after retest).
- By Operator (Factor II & Factor V): Overall total agreement by operator was 99.7% (299/300).
  - Operators at Site 1 & 2: 100% (50/50 each).
  - Operators at Site 3: OP 1 100% (50/50), OP 2 98.0% (49/50) (One sample indeterminate after retest).
- By Lot (Factor II & Factor V): Overall total agreement by lot was 100% (150/150).

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

Overall accuracy (agreement) relative to bi-directional sequencing: 99.3% for both Factor II and Factor V.
Reproducibility: Overall total agreement by site 99.7%, by operator 99.7%, by lot 100%.

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.

Roche Factor II (Prothrombin) G20210A Kit (510(k) #K033612), Roche Factor V Leiden Kit (510(k) #K033607)

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

§ 864.7280 Factor V Leiden DNA mutation detection systems.

(a)
Identification. Factor V Leiden deoxyribonucleic acid (DNA) mutation detection systems are devices that consist of different reagents and instruments which include polymerase chain reaction (PCR) primers, hybridization matrices, thermal cyclers, imagers, and software packages. The detection of the Factor V Leiden mutation aids in the diagnosis of patients with suspected thrombophilia.(b)
Classification. Class II (special controls). The special control is FDA's guidance entitled “Class II Special Controls Guidance Document: Factor V Leiden DNA Mutation Detection Systems.” (See § 864.1(d) for the availability of this guidance document.)

Summary

Cepheid.

SFP 1 8 2009

510(k) Summary

As required by 21 CFR Section 807.92(c).

Submitted by:

Cepheid® 904 Caribbean Drive Sunnyvale, CA 90489 Phone number: (408) 400-8230 Fax number: (408) 541-6439

Xpert® HemosIL® FII & FV

Factor II and Factor V Genotyping Assay.

Xpert® HemosIL® Factor II & Factor V

Xpert® HemosIL® Factor II & Factor V Assay

Nucleic Acid Amplification Test, DNA, Factor II Prothrombin G20210A and Factor V G1691A (Leiden)

Factor II Prothrombin and Factor V Leiden DNA Mutation

Russel K. Enns, Ph.D.

qualitative, genotyping

July 31, 2009

Contact: Date of Preparation:

Device:

Trade name:

Common names:

Type of Test:

Classification name:

Advisory Committee:

Detection Systems

Regulation number: 864.7280

NPR

Classification Hematology

Predicate Devices:

Procode:

Roche Factor II (Prothrombin) G20210A Kit (510(k) #K033612) and Roche Factor V Leiden Kit (510(k) #K033607)

Device Description:

Xpert HemosIL Factor II & Factor V Assay 510(k) Summary

Page 1 of 10

K682118

sample preparation, amplification, and real-time detection are all fully-automated and completely integrated.

The test includes a Probe Check Control (PCC) that verifies reagent rehydration, PCR tube filling in the cartridge, probe integrity, and dye stability and an instrument control verifies the instrument is performing properly. Additionally, the sample functions as its own internal control since both normal and mutant gene sequences are detected, and each person.tested is expected to have one of these sequence signatures.

Device Intended Use:

The Xpert™ HemosIL® Factor II & Factor V Assay is a qualitative in vitro diagnostic genotyping test for the detection of Factor II and Factor V alleles from sodium citrate or EDTA anticoagulated whole blood. The assay is performed on the Cepheid GeneXpert® Dx System. This test is intended to provide results for Factor II (G20210A) and Factor V Leiden (G1691A) mutations as an aid in the diagnosis in individuals with suspected thrombophilia.

Substantial Equivalence:

The Xpert HemosIL Factor II & Factor V Assay is substantially equivalent to two predicate devices. Roche Factor II (Prothrombin) G20210A Kit (510(k) #K033612) and Roche Factor V Leiden Kit (510(k) #K033607). The Xpert HemosIL Factor II & Factor V Assay and Roche Factor II (Prothrombin) G20210A Kit and Factor V Leiden Kit determine the Factor II and Factor V genotypes through real-time PCR amplification and fluorogenic target-specific hybridization detection.

Table 5.1 shows the similarities and differences between the Xpert HemosIL Factor II & Factor V Assay and the two predicate devices. The GeneXpert Dx System, used with the Xpert HemosIL Factor II & Factor V Assay, is also used to perform the Cepheid Xpert GBS Assay, the Xpert EV Assay and the Xpert MRSA Assay. The predicate devices are performed on the Roche LightCycler® system.

In a multi-center clinical comparison study, performance of the Xpert HemosIL Factor II & Factor V Assay and the Roche Factor II and Factor V Assays were each calculated relative to the gold standard, bi-directional sequencing results, and were shown to be substantially equivalent.

Table 5.1

Similarities and Differences Between the Xpert HemosIL Factor II & Factor V Assay and the Predicate Devices

Similarities (Assay)
Item	Device	Predicates
	Xpert HemosIL Factor II &
Factor V Assay	Roche Factor II
(Prothrombin)
G20210A
(510(k) #K033612)	Roche Factor
V Leiden Kit
(510(k) #K033607)
Intended Use	Rapid detection of Factor II
and Factor V alleles from
sodium citrate and EDTA
anticoagulated whole blood	Same except
detection of
Factor II only
from EDTA
anticoagulated
blood only	Same except
detection of
Factor V only
from EDTA
anticoagulated
blood only
Indication for Use	Aid in the diagnosis in
individuals with suspected
thrombophilia	Same	Same
Technological
Detection
Principles	Amplification and detection
system for nucleic acids using
fluorescence detection.	Same	Same

Differences (Assay)
Item	Device	Predicates
	Xpert HemosIL Factor II &
Factor V Assay	Roche Factor II
(Prothrombin)
G20210A
(510(k) #K033612)	Roche Factor
V Leiden Kit
(510(k) #K033607)
Specimen Type	Anticoagulated Whole Blood	Purified DNA
from human
blood samples	Purified DNA
from human
blood samples
Sample
Preparation	Automated On-line	Performed off-
line	Performed off-
line
Test Cartridge	Disposable single-use, multi-
chambered fluidic cartridge.	Disposable
single-use PCR
capillary	Disposable
single-use PCR
capillary

Differences (Assay)
Item	Device	Predicates
	Xpert HemosIL Factor II &
Factor V Assay	Roche Factor II
(Prothrombin)
G20210A
(510(k) #K033612)	Roche Factor
V Leiden Kit
(510(k) #K033607)
Instrument System	Cepheid GeneXpert® Dx
System	Roche
LightCycler	Roche
LightCycler
Detection
Chemistry	Paired hybridization probes
using Scorpions	Paired
hybridization
probes using
fluorescence
energy transfer
(FRET)	Paired
hybridization
probes using
fluorescence
energy transfer
(FRET)
Fluidics/Sample
Preparation	Self-contained and automated
after two single-dose reagent
additions.	Manual	Manual
Probes	Scorpion Probes	HybProbe	HybProbe
Controls	Internal Probe check control
(PCC).	External positive
and negative
controls required
per run	External positive
and negative
controls required
per run
Rapid test results	Approximately 30 minutes to
results.