Hepatitis C Antibody with Reflex to PCR

The Qualitative detection of Hepatitis C virus IgG and IgM antibodies (anti-HCV) in human sera by the FDA approved Abbott ARCHITECT Anti-HCV test two-step chemiluminescent immunoassay.

In the first step, sample, assay diluent, and recombinant HCV antigen (rHCV) coated paramagnetic microparticles are combined. Anti-HCV present in the sample binds to the rHCV coated microparticles. In the second step, anti-human IgG/IgM acridinium-labeled conjugate is added, which binds to IgG and IgM anti-HCV. Then pre-trigger and trigger solutions are added to the reaction mixture. The resulting chemiluminescent reaction is measured as relative light units (RLUs).

The presence or absence of IgG/IgM anti-HCV in the sample is determined by comparing the chemiluminescent signal in the reaction to the cutoff signal determined from an ARCHITECT Anti-HCV calibration. Specimens with signal to cutoff (S/CO) values ≥ 1.00 are considered reactive for IgG/IgM anti-HCV. Specimens with S/CO values < 0.79 are considered nonreactive and specimens with S/CO values between 0.80 and 0.99 are Indeterminate.

Reactive anti-HCV will reflex to Hepatitis C RNA, Quantitative [HCVQNT] for confirmation with an additional charge.

For anti-HCV testing without PCR reflex for REACTIVE results, see Hepatitis C Antibody without PCR reflex on reactive samples [HCABX].

All Locations: 2 tubes required if reflex testing is ordered

Preferred: 5 mL blood in GOLD SST tube plus 5 mL blood in PEARL PPT tube

Also acceptable: 5 mL blood in ORANGE RST or RED TOP tube plus a 5 mL PEARL PPT tube for reflexive testing.

Note: If no PEARL PPT is received, any reflexive molecular testing can be done on GOLD SST, LAVENDER or RED TOP Tube that has not been on automated lines.

GOLD SST tube: Specimens may be stored on or off the clot, red blood cells, or separator gel for up to 3 days at room temperature (15-30°C) or up to 7 days at 2-8°C. If testing will be delayed more than 3 days for specimens stored at room temperature or more than 7 days for specimens stored at 2-8°C, remove serum from the clot, red blood cells, or separator gel and store at -20°C or colder. Avoid more than three freeze/thaw cycles.

PEARL PPT tube: centrifuge within 6 hours of collection and freeze spun PEARL PPT tube at -20°C. Route frozen PEARL tubes to UW SPS -20°C freezer rack 80. [ok to freeze spun pearl top tubes on gel.]

Unacceptable samples: Cord blood, neonatal specimens, cadaver specimens, heat-inactivated specimens, or body fluids such as saliva, urine, amniotic fluid, or pleural fluid. Specimens with obvious microbial contamination or are grossly hemolyzed.