Document Detail


Determination of HIV-1 subtypes (A-D, F, G, CRF01_AE) by PCR in the transmembrane region (gp41) with novel primers.
MedLine Citation:
PMID:  15778948     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
HIV-1 has a huge genetic diversity. So far, nine subtypes have been isolated, namely, subtypes A, B, C, D, F, G, H, J, and K. Epidemiological study provides information which may help in the development of HIV-1 prevention programs or health policies. In the future, subtyping may also be critical for vaccine development, and an effective anti-viral drug will need to be effective for different subtypes of HIV virus. The analysis of the nucleotide sequence of the v3 region is considered the most reliable method for determining the HIV-1 subtype. However, the procedures for determining the v3 sequences are complicated and time consuming, requiring expensive reagents, equipment, and well-trained personnel. The polymerase chain reaction (PCR) method using subtype-specific primers for HIV-1 subtyping is easier and faster. The objective of this study was to develop subtype-specific primers for subtyping PCR. The specific primers were designed for subtypes A, B, C, D, F, G, and CRF01_AE, and these primers could be applied to assay for various HIV-1 subtypes in the clinical samples. The specific primers were designed for each subtypes in the gp41 region. The result of PCR was compared with the subtypes which was determined by the v3 sequence. The results of subtyping by PCR using the newly designed primers could detect 29 of 33 patients tested, and all matched those obtained by nucleotide sequencing of the env v3 region except for three subjects, which were differentiated as CRF02_AG. The newly designed primers functioned accurately and conclusively. In comparison with PCR as a method for the determination of subtypes, sequence analysis requires better-trained personnel, more expensive reagents, and more equipment and time.
Authors:
Fumihiro Yagyu; Shoko Okitsu; Kenichi Tanamoto; Hiroshi Ushijima
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of medical virology     Volume:  76     ISSN:  0146-6615     ISO Abbreviation:  J. Med. Virol.     Publication Date:  2005 May 
Date Detail:
Created Date:  2005-03-28     Completed Date:  2005-08-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7705876     Medline TA:  J Med Virol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  16-23     Citation Subset:  IM    
Copyright Information:
Copyright 2005 Wiley-Liss, Inc.
Affiliation:
Department of Developmental Medical Sciences, Institute of International Health, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
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MeSH Terms
Descriptor/Qualifier:
Adolescent
Adult
Base Sequence
DNA Primers
Female
HIV Envelope Protein gp120 / genetics
HIV Envelope Protein gp41 / genetics*
HIV Infections / diagnosis
HIV-1 / classification,  genetics*
Humans
Male
Middle Aged
Peptide Fragments / genetics
Polymerase Chain Reaction / economics,  methods*
Chemical
Reg. No./Substance:
0/DNA Primers; 0/HIV Envelope Protein gp120; 0/HIV Envelope Protein gp41; 0/HIV envelope protein gp120 (305-321); 0/Peptide Fragments

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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