NAME

Abbott RealTime HCV Genotype II

INTENDED USE

The Abbott RealTime HCV Genotype II is an in vitro reverse transcription-polymerase chain reaction

(RT-PCR) assay for use with the Abbott mSample Preparation System reagents and with the Abbott m2000sp and m2000rt instruments for the qualitative identification of hepatitis C virus (HCV) genotypes 1, 1a, 1b, and 2 - 5 in plasma or serum from individuals chronically infected with HCV.

The Abbott RealTime HCV Genotype II is intended for use as an aid in the management of HCV-infected individuals and in guiding the selection of therapeutic treatment indicated for the above listed genotypes. The assay is intended for use on patients who are chronically infected with HCV, are being considered for antiviral treatment, and are positive for HCV RNA.

The Abbott RealTime HCV Genotype II assay is not for screening blood, plasma, serum or tissue donors for HCV.

SUMMARY AND EXPLANATION OF THE TEST

The Hepatitis C virus (HCV), a significant cause of blood-borne hepatitis, is an enveloped virus containing a single-stranded positive sense RNA genome of approximately 9,500 nucleotides.1 It has been identified as the major etiological agent for post-transfusion non-A and non-B hepatitis worldwide. Based on genetic similarity, HCV has been classified into six major genotypes (1 – 6) and numerous subtypes (1a, 1b, etc.).2 HCV genotype impacts the response of HCV-infected patients to peg-interferon/ribavirin combination therapy.3 Before starting combination therapy, it is recommended that the genotype of the infecting HCV isolate be determined so that the patient can receive the most appropriate therapy regimen.4

HCV genotype 1 is the predominant HCV genotype in the United States (73.32%) followed by HCV genotype 2 (13.10%) and HCV genotype 3 (12.13%) and HCV genotype 4 (1.32%). HCV genotypes 5 and 6 each represent less than or equal to 1% of HCV genotype determinations in the US.5

The Abbott RealTime HCV Genotype II Positive Control is standardized against the Second WHO International Standard for Hepatitis C Virus RNA (NIBSC Code 96/798).6

BIOLOGICAL PRINCIPLES OF THE PROCEDURE

The Abbott RealTime HCV Genotype II assay consists of two reagent kits:

Abbott RealTime HCV Genotype II Amplification Reagent Kit

Abbott RealTime HCV Genotype II Control Kit

The Abbott RealTime HCV Genotype II is an in vitro reverse transcription-polymerase chain reaction

(RT-PCR) assay for the identification of the genotype(s) of hepatitis C virus (HCV) in plasma or serum from individuals chronically infected with HCV. The Abbott RealTime HCV Genotype II assay detects genotypes 1, 1a, 1b and 2 – 5 through the use of genotype-specific fluorescent-labeled oligonucleotide probes.

Sample Preparation

The Abbott m2000sp provides automated sample preparation using a magnetic microparticle-based protocol (Abbott mSample Preparation System) to process 0.5 mL samples (ACD-A, CPD, potassium EDTA, or sodium EDTA plasma or serum). During the sample preparation protocol, HCV virions are disrupted by guanidine isothiocyanate, RNA is captured on the magnetic microparticles, inhibitors are removed by washing steps, and RNA is eluted off the microparticles. The bound nucleic acids are eluted and transferred to a 96 deep-well plate. The nucleic acids are then ready for amplification. The Internal Control (IC) is introduced into each specimen at