Rapid HCV RNA detection by PCR followed by a new non-radioactive liquid hybridisation assay and comparison with RIBA.
A one-stage polymerase chain reaction (PCR) followed by an automated liquid hybridisation assay was used to examine anti-HCV-positive patients. The presence of HCV-RNA in 251 randomly selected enzyme immunoassay (EIA) positive clinical specimens was compared to their RIBA pattern. An association of a RIBA pattern with presence or absence of HCV-RNA was not detected. One hundred of these samples were also evaluated after dividing them into normal and elevated serum alanin aminotransferase (ALT) levels. PCR results were obtained on the basis of amplification products from two different gene regions (5'-NC region and NS 3). To prove the specificity of the PCR products, a commercially available digoxigenin-based liquid hybridisation assay was evaluated. The sensitivity was comparable to the results obtained after nested PCR. Based on the results of the study, the two-stage PCR can be changed in favour of the easier one-step PCR which offers the advantage of fewer contamination problems.
Schacker
U
U
Medical Diagnostic Laboratory, Baden-Baden, Germany.
Bermayer
H P
HP
Boehler
V
V
Ionescu
D
D
Zapata
M
M
Grauer
H
H
Rai
K
K
D000410:Alanine Transaminase / Q000097:blood; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000138:chemical synthesis; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000097:blood / Q000821:virology; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral; D000410:Alanine Transaminase