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Rapid HCV RNA detection by PCR followed by a new non‐radioactive liquid hybridisation assay and comparison with RIBA
Author(s) -
Schacker Ulrike,
Bermayer HansPeter,
Boehler Valerie,
Ionescu Doina,
Zapata Manuel,
Grauer Hasan,
Rai Kamal
Publication year - 1995
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890460403
Subject(s) - microbiology and biotechnology , polymerase chain reaction , virology , immunoassay , biology , digoxigenin , rna , hepatitis c virus , virus , nested polymerase chain reaction , chemistry , chromatography , gene , messenger rna , in situ hybridization , immunology , biochemistry , antibody
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. © 1995 Wiley‐Liss, Inc.