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Typing of hepatitis C virus isolates by DNA enzyme immunoassay

Viazov, Sergei; Zibert, Andree; Ramakrishnan, Kandiah; Widell, Anders LU ; Cavicchini, Ada; Schreier, Eckart and Roggendorf, Michael (1994) In Journal of Virological Methods 48(1). p.81-91
Abstract
Recently, at least six types of hepatitis C viruses (HCV) have been identified. Different types of HCV appear to possess different pathogenic properties and a different sensitivity to interferon treatment. Typing of HCV isolates may therefore be an important diagnostic procedure. We report on a new method for identification of HCV types 1a, 1b, 2a, 2b and 3a which are most prevalent in Europe, North America and Japan. The assay is based on a combination of two well established techniques, the polymerase chain reaction (PCR) and DNA enzyme immunoassay (DEIA). In the first step of the method a cDNA of about 250 bp corresponding to the HCV core-region is amplified by nested PCR. The target cDNA is then hybridized to type-specific... (More)
Recently, at least six types of hepatitis C viruses (HCV) have been identified. Different types of HCV appear to possess different pathogenic properties and a different sensitivity to interferon treatment. Typing of HCV isolates may therefore be an important diagnostic procedure. We report on a new method for identification of HCV types 1a, 1b, 2a, 2b and 3a which are most prevalent in Europe, North America and Japan. The assay is based on a combination of two well established techniques, the polymerase chain reaction (PCR) and DNA enzyme immunoassay (DEIA). In the first step of the method a cDNA of about 250 bp corresponding to the HCV core-region is amplified by nested PCR. The target cDNA is then hybridized to type-specific oligonucleotides fixed to a solid phase through an avidin-biotin bridge. The formed hybrids are detected by a standard ELISA using monoclonal antibodies reacting with double-stranded DNA. Typically, signal-to-noise (S/N) ratios between 18.2 and 48.6 could be observed when different HCV types/subtypes were analyzed by this method. The test was evaluated using cloned HCV cDNAs of known types and by sequence determination of some of the typed cDNAs. Typing of 115 isolates from Germany, Russia and Turkey revealed that subtype 1b (59-100%) and 1a (24-32%) are most prevalent in these countries. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Hepatitis C virus, Genotype, DNA enzyme immunoassay, Polymerase chain reaction
in
Journal of Virological Methods
volume
48
issue
1
pages
81 - 91
publisher
Elsevier
external identifiers
  • pmid:7962263
  • scopus:0028167889
ISSN
1879-0984
DOI
10.1016/0166-0934(94)90091-4
language
English
LU publication?
yes
id
c9c80d0c-a504-4b82-87a7-6fa04f41f1b3 (old id 1107749)
date added to LUP
2008-07-22 14:38:24
date last changed
2017-08-06 03:38:40
@article{c9c80d0c-a504-4b82-87a7-6fa04f41f1b3,
  abstract     = {Recently, at least six types of hepatitis C viruses (HCV) have been identified. Different types of HCV appear to possess different pathogenic properties and a different sensitivity to interferon treatment. Typing of HCV isolates may therefore be an important diagnostic procedure. We report on a new method for identification of HCV types 1a, 1b, 2a, 2b and 3a which are most prevalent in Europe, North America and Japan. The assay is based on a combination of two well established techniques, the polymerase chain reaction (PCR) and DNA enzyme immunoassay (DEIA). In the first step of the method a cDNA of about 250 bp corresponding to the HCV core-region is amplified by nested PCR. The target cDNA is then hybridized to type-specific oligonucleotides fixed to a solid phase through an avidin-biotin bridge. The formed hybrids are detected by a standard ELISA using monoclonal antibodies reacting with double-stranded DNA. Typically, signal-to-noise (S/N) ratios between 18.2 and 48.6 could be observed when different HCV types/subtypes were analyzed by this method. The test was evaluated using cloned HCV cDNAs of known types and by sequence determination of some of the typed cDNAs. Typing of 115 isolates from Germany, Russia and Turkey revealed that subtype 1b (59-100%) and 1a (24-32%) are most prevalent in these countries.},
  author       = {Viazov, Sergei and Zibert, Andree and Ramakrishnan, Kandiah and Widell, Anders and Cavicchini, Ada and Schreier, Eckart and Roggendorf, Michael},
  issn         = {1879-0984},
  keyword      = {Hepatitis C virus,Genotype,DNA enzyme immunoassay,Polymerase chain reaction},
  language     = {eng},
  number       = {1},
  pages        = {81--91},
  publisher    = {Elsevier},
  series       = {Journal of Virological Methods},
  title        = {Typing of hepatitis C virus isolates by DNA enzyme immunoassay},
  url          = {http://dx.doi.org/10.1016/0166-0934(94)90091-4},
  volume       = {48},
  year         = {1994},
}