Hepatitis C RNA Genotyping by PCR

The Abbott real-time FDA approved HCV assay genotypes the Hepatitis C virus by real-time PCR amplification. The six major HCV genotypes have at least one genotype-specific single-nucleotide polymorphism in the 5' non-coding region HCV RNA in the patient’s sample is amplified with 9 separate allele-specific primer/probe sets at the 5’ UTR and NS5b regions of the virus. RNA is added by the instrument into 3 different mixes (A, B, and C) using the Genotype II kit. Based on which genotype(s) is/are present, there will be signal from the probe in either A, B, or C, or a combination. There are six major genotypes of HCV, with numerous characterized subtypes 1, 2, 3, 4, 5, and 6. The RNA concentration detected with a probability of 95% or greater of the Genotype assay is 500 IU/mL (2.70 log IU/mL). About 7% of samples are unable to be genotyped, either due to the lack of specific signal or to the interpretation of a mixed signal. These samples will require Sanger sequencing of the amplicon to determine the genotype.

For HCV quantitation, see Hepatitis C RNA, Quantitative [HCVQNT].

Blood: 5 mL blood in PPT (PEARL TOP) tube.

Also Acceptable: 5 mL blood in GOLD SST, RED, or EDTA (LAVENDAR TOP) tube.

Unacceptable samples: Plasma collected in sodium or lithium heparin.

Note: Separate specimens MUST be submitted when both Viral Culture/Serology and PCR testing are ordered.

Blood: Separate serum or plasma from whole blood within 6 hours of collection. Freeze plasma at -10°C or colder and ship on dry ice. Serum or plasma may be stored at 2-8°C (ice pack) for up to 48 hours from collection. Do not freeze/thaw.

Outside Laboratory: Ship samples on dry ice. If freezer and/or dry ice is not available, ship immediately at 2-8°C (ice pack). Shipment time must not exceed 48 hours from collection. Sample may be stored at 2-8°C (ice pack) for up to 48 hours of collection. Samples stored at 2-8°C greater than 48 hours old may be rejected. Do not freeze/thaw.