Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol
In the past few years, many new subtypes in hepatitis C virus (HCV) genotype 6 have been identified. The aim of this study was to modify the multiplex real-time polymerase chain reaction (RT-PCR) protocol and use it to determine the HCV subtypes of a group of Taiwanese injection drug users (IDUs). M...
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| Format: | Article |
| Language: | English |
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Elsevier
2011-12-01
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| Series: | Journal of the Formosan Medical Association |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0929664611001318 |
| _version_ | 1811237610052386816 |
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| author | Yuan-Ming Lee Yen-Ju Chen Cheng-Ming Lee Lou-Hui Kuo Wing-Wai Wong Yi-Ming Arthur Chen |
| author_facet | Yuan-Ming Lee Yen-Ju Chen Cheng-Ming Lee Lou-Hui Kuo Wing-Wai Wong Yi-Ming Arthur Chen |
| author_sort | Yuan-Ming Lee |
| collection | DOAJ |
| description | In the past few years, many new subtypes in hepatitis C virus (HCV) genotype 6 have been identified. The aim of this study was to modify the multiplex real-time polymerase chain reaction (RT-PCR) protocol and use it to determine the HCV subtypes of a group of Taiwanese injection drug users (IDUs).
Methods: We used 76 serum specimens collected in northern Taiwan in 2008. Multiplex RT-PCR was used for HCV subtyping among those serum samples having anti-HCV antibodies. Twenty cases were randomly selected for comparison with subtyping results from Inno-LiPa II tests and phylogenetic tree analysis using NS5B sequences.
Results: Multiplex RT-PCR assays showed that 60.5% (46/76) of IDUs had single HCV infection. Three out of 76 (3.9%) had double HCV infection (1b/6a, 2a/2b and 2b/6a). Besides this, 27.6% (21/76) had no HCV signal. One IDU had subtype 6n and two had subtype 6w infection. Inno-LiPa II tests misclassified all 6n and 6w cases as 1b subtype.
Conclusion: Our modified multiplex RT-PCR protocol can be used to support molecular epidemiological studies and laboratory diagnoses of different HCV subtypes including genotype 6. |
| first_indexed | 2024-04-12T12:26:14Z |
| format | Article |
| id | doaj.art-688d898d636745b5a6f7f808a7fd77aa |
| institution | Directory Open Access Journal |
| issn | 0929-6646 |
| language | English |
| last_indexed | 2024-04-12T12:26:14Z |
| publishDate | 2011-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of the Formosan Medical Association |
| spelling | doaj.art-688d898d636745b5a6f7f808a7fd77aa2022-12-22T03:33:09ZengElsevierJournal of the Formosan Medical Association0929-66462011-12-011101276276710.1016/j.jfma.2011.11.006Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocolYuan-Ming Lee0Yen-Ju Chen1Cheng-Ming Lee2Lou-Hui Kuo3Wing-Wai Wong4Yi-Ming Arthur Chen5AIDS Prevention and Research Center, National Yang-Ming University, Taipei, TaiwanAIDS Prevention and Research Center, National Yang-Ming University, Taipei, TaiwanAIDS Prevention and Research Center, National Yang-Ming University, Taipei, TaiwanAIDS Prevention and Research Center, National Yang-Ming University, Taipei, TaiwanAIDS Prevention and Research Center, National Yang-Ming University, Taipei, TaiwanAIDS Prevention and Research Center, National Yang-Ming University, Taipei, TaiwanIn the past few years, many new subtypes in hepatitis C virus (HCV) genotype 6 have been identified. The aim of this study was to modify the multiplex real-time polymerase chain reaction (RT-PCR) protocol and use it to determine the HCV subtypes of a group of Taiwanese injection drug users (IDUs). Methods: We used 76 serum specimens collected in northern Taiwan in 2008. Multiplex RT-PCR was used for HCV subtyping among those serum samples having anti-HCV antibodies. Twenty cases were randomly selected for comparison with subtyping results from Inno-LiPa II tests and phylogenetic tree analysis using NS5B sequences. Results: Multiplex RT-PCR assays showed that 60.5% (46/76) of IDUs had single HCV infection. Three out of 76 (3.9%) had double HCV infection (1b/6a, 2a/2b and 2b/6a). Besides this, 27.6% (21/76) had no HCV signal. One IDU had subtype 6n and two had subtype 6w infection. Inno-LiPa II tests misclassified all 6n and 6w cases as 1b subtype. Conclusion: Our modified multiplex RT-PCR protocol can be used to support molecular epidemiological studies and laboratory diagnoses of different HCV subtypes including genotype 6.http://www.sciencedirect.com/science/article/pii/S0929664611001318hepatitis C virus (HCV)genotype 6injection drug usersInno-LiPa II testmultiplex RT-PCRsubtype 6nsubtype 6w |
| spellingShingle | Yuan-Ming Lee Yen-Ju Chen Cheng-Ming Lee Lou-Hui Kuo Wing-Wai Wong Yi-Ming Arthur Chen Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol Journal of the Formosan Medical Association hepatitis C virus (HCV) genotype 6 injection drug users Inno-LiPa II test multiplex RT-PCR subtype 6n subtype 6w |
| title | Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol |
| title_full | Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol |
| title_fullStr | Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol |
| title_full_unstemmed | Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol |
| title_short | Detection of hepatitis C virus subtypes 6a, 6n, 6w and mixed infections using a modified multiplex real-time polymerase chain reaction protocol |
| title_sort | detection of hepatitis c virus subtypes 6a 6n 6w and mixed infections using a modified multiplex real time polymerase chain reaction protocol |
| topic | hepatitis C virus (HCV) genotype 6 injection drug users Inno-LiPa II test multiplex RT-PCR subtype 6n subtype 6w |
| url | http://www.sciencedirect.com/science/article/pii/S0929664611001318 |
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