Typing of hepatitis C virus isolates by DNA enzyme immunoassay
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1107749
- author
- Viazov, Sergei ; Zibert, Andree ; Ramakrishnan, Kandiah ; Widell, Anders LU ; Cavicchini, Ada ; Schreier, Eckart and Roggendorf, Michael
- organization
- publishing date
- 1994
- 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
- 2016-04-01 11:59:19
- date last changed
- 2021-01-03 04:02:37
@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}},
keywords = {{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}},
doi = {{10.1016/0166-0934(94)90091-4}},
volume = {{48}},
year = {{1994}},
}