RealTime HCV

1105_20348
RealTime HCV

Description

The Abbott RealTime HCV assay is an in vitro reverse transcription-polymerase chain reaction (RT-PCR) assay for the quantitation of hepatitis C viral ribonucleic acid (HCV RNA) in human serum and plasma from HCV-infected individuals. The Abbott RealTime HCV assay is intended for use as an aid in the management of HCV-infected patients under going antiviral therapy.

Abbott RealTime HCV Performance *

Sensitivity 12 IU/mL (0.5 mL input); 30 IU/mL (0.2 mL input)
Linear Range 12 IU/mL (1.08 log IU/mL) to 100 million IU/mL (log 8.0 IU/mL)
Precision Inter-assay SD at 5.97 log IU/mL = 0.04, Inter-assay SD at 1.96 log IU/mL = 0.09
Specificity >= 99.5%†
Genotype Detection Equal quantification of genotype 1, 2, 3, 4, 5 and 6
Standardization Second WHO international standard for HCV RNA
Internal Control Added to lysis buffer during extraction and detected at all levels

*Performance data shown from internal verification. The Abbott RealTimeHCV assay is not for screening blood, plasma, serum or tissue donorsfor HCV, or to be used as a diagnostic test to confirm the presence ofHCV infection.

†The specificity of the RealTime HCV assay wasevaluated by testing 56 HCV seronegative serum and 56 HCV seronegativeplasma specimens. HCV RNA was not detected, resulting in 100 %(112/112) specificity in this representative study.

Detection and Quantitation of HCV Genotypes
Genotype dilution linearity was demonstrated by diluting six specimens, one of each genotype 1 through 6, to target concentrations of 5.0 log IU/ml,3.5 log IU/ml, 2.0 log IU/ml, and 1.0 log IU/ml. Four replicates were tested at each concentration of each genotype. The correlation coefficients ranged from 0.994 to 0.998.


Correlation to Comparator Assay
Specimens from 131 HCV infected patients were tested with the RealTime HCV assay and a comparator assay per NCCLS Document EP9-A2.

Design

Development Philosophy
Today's clinical molecular diagnostics laboratory must have confidence in the quality of HCV patient results. As a result of our real-time PCR development philosophy, Abbott RealTime HCV is engineered to quantify all HCV genotypes (1—6) equally.

Accurate Quantification is dependent upon a combination of:
  • Primer Design
  • Probe Design
  • Cycling Conditions

Primers and Probe are Targeted to the 5´UTR Region of the HCV Genome
The target sequence for the Abbott RealTime HCV assay is in the 5´utr region of the HCV genome. This region is specific for HCV and is highly conserved. The primers are designed to hybridize to the 5´utr regionwith the fewest possible mismatches among HCV genotypes 1-6.

Single Stranded Linear Probe Design

In the absence of target, the probe achieves quenching through random coiling.


In presence of target, the probe prefers to hybridize to the target sequence, allowing fluorescent detection; cleavage is not required.

Cycling Conditions:  Low Temperature Read Cycles

The cycling conditions for Abbott RealTime HCV encompass a low temperature read cycle. This read cycle allows the probe to tolerate mismatches more effectively than a probe required to bind during the extension phase.

Order Information

Product Name Package Size Vysis® Order Number Abbott Order Number
RealTime HCV Abbott RealTime HCV Calibrator Kit 12 Cal A, 12 Cal B (4 Complete Calibration Sets) 04J86-70 04J86-70
RealTime HCV Abbott RealTime HCV Control Kit 8 Low Positive, 8 High Positive, 8 Negative 04J86-80 04J86-80
RealTime HCV Abbott RealTime HCV Amplification Reagent Kit 96 Assays (4 packs x 24 assays) 04J86-90 04J86-90
RealTime HCV Abbott RealTime HCV Application CD-ROM 01L69 01L69
In Vitro Diagnostic Available in the United States.
 
CE Marked