RealTime HCV Genotype II

Description

Intended Use

The Abbott RealTime HCV Genotype II is an in vitro reverse transcription-polymerase chain reaction (RT-PCR) assay for determining the genotype(s) of hepatitis C virus (HCV) in plasma and serum from HCV-infected individuals. The Abbott RealTime HCV Genotype II is not for screening blood, plasma serum or tissue donors for HCV, or to be used as a diagnostic test to confirm the presence of HCV infection in donated blood, plasma, serum or tissue.

Abbott RealTime HCV Genotype performance*

Sensitivity		500 IU/mL 0.5 mL prep.; 1250 IU/mL 0.2 mL prep.
Specificity		100% **
Target Region		5'UTR & NS5b
Genotype Detection		1a (NS5b), 1b (NS5b), 1, 2, 3, 4, 5, 6 (5'UTR)
Standardization		Second WHO International Standard for Hepatitis C Virus RNA
Internal Control		Yes; processed through sample prep with each sample
External Control		Yes; Negative, Positive
Results reported		Qualitative - Genotype Call

*Performance data shown from internal verification.

**The specificity of Abbott RealTime Genotype II was evaluated by testing 100 HCV serologically negative plasma specimens. All negative specimens tested were interpreted as HCV RNA negative by the Abbott RealTime HCV Genotype II assay, resulting in 100% specificity.

Design

Target Region
The Abbott RealTime HCV Genotype II assay detects genotypes 1, 2, 3, 4, 5, and 6, and subtypes 1a and 1b through the use of genotype-specific fluorescent-labeled oligonucleotide probes. It targets the 5'UTR for the classification of HCV, HCV genotypes 1, 2, 3, 4, 5, and 6, and the NS5B region to accurately subtype HCV genotypes 1a and 1b.

The accuracy of the Abbott RealTime HCV Genotype II assay was demonstrated by testing 169 HCV genotype 1 (110 of genotype 1a, 58 of subtype 1b, and 1 genotype 1 only), 41 HCV genotype 2, 27 HCV genotype 3, 18 HCV genotype 4, 14 HCV genotype 5, and 12 HCV genotype 6 positive specimens. Nucleotide sequencing was used to determine the reference genotype of each specimen tested.

HCV Genotype/Subtype	Number Samples Tested	Number Samples Correctly Identified	Detection Rate (%)
1*	169	169	100.00
1a**	110	107	97.27
1b***	58	56	96.55
2	41	41	100.00
3	27	26	96.30
4	18	18	100.00
5	14	14	100.00
6	12	8	66.67
1 through 6‡	281	276	98.22

• * Denotes analysis based upon samples tested with m2000rt results: 1, 1a, 1b, 1/1a, and 1/1b.
• ** Three out of 110 subtype 1a samples were identified as genotype 1 only but not as subtype 1a.
• *** One site identified subtype 1b sample was excluded from analysis because it was identified by sequencing as genotype 1 only. Two out of 58 remaining subtype 1b samples were identified as genotype 1 but not as subtype 1b.
• ‡ Denotes analysis based upon genotypes 1, 2, 3, 4, 5, and 6 combined.

Detection
The assay requires three separate reactions to detect genotypes 1,2,3,4,5, and 6 and subtypes 1a and 1b:

• Reaction A is designed to detect all HCV isolates, subtype 1a isolates, and genotype 3 isolates.
• Reaction B is designed to detect genotype 2 isolates, subtype 1b isolates, and genotype 1 isolates.
• Reaction C is designed to detect genotype 4 isolates, genotype 5 isolates, and genotype 6 isolates.