Document Detail


A novel multiplex RT-PCR for identification of VP6 subgroups of human and porcine rotaviruses.
MedLine Citation:
PMID:  20546787     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
VP6 protein antigens allow classification of rotaviruses into at least four subgroups, depending on the presence or absence of SG-specific epitopes: SG I, SG II, SG (I+II), and SG non-(I+II). However, MAbs against epitopes on the VP6 protein of human and porcine rotaviruses, sometimes, do not recognize SG-specific epitopes or recognize irrelevant-SG epitopes and therefore result in the incorrect assignment of subgroups. In order to solve this problem, a novel multiplex RT-PCR was developed as an alternative tool to identify VP6 genogroups using newly designed primers which are specific for genogroup I or II. The sensitivity and specificity of the newly developed multiplex RT-PCR method was evaluated by testing with human and porcine rotaviruses of known SG I, SG II, SG (I+II), and SG non-(I+II) strains in comparison with monoplex RT-PCR and VP6 sequence analysis. The results show that the genogroups of both human and porcine rotaviruses as determined by the new multiplex RT-PCR method were in 100% agreement with those determined by monoplex RT-PCR and VP6 sequence analysis. The method was shown to be specific, sensitive, less-time consuming, and successful in genogrouping clinical isolates of rotaviruses circulating in children and piglets with acute diarrhea.
Authors:
Aksara Thongprachum; Natthawan Chaimongkol; Pattara Khamrin; Chansom Pantip; Masashi Mizuguchi; Hiroshi Ushijima; Niwat Maneekarn
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-05-28
Journal Detail:
Title:  Journal of virological methods     Volume:  168     ISSN:  1879-0984     ISO Abbreviation:  J. Virol. Methods     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-07-26     Completed Date:  2010-11-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8005839     Medline TA:  J Virol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  191-6     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Elsevier B.V. All rights reserved.
Affiliation:
Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, Viral / genetics*
Capsid Proteins / genetics*
Child
Child, Preschool
DNA Primers / genetics
Feces / virology
Humans
Molecular Epidemiology / methods
Reverse Transcriptase Polymerase Chain Reaction / methods*
Rotavirus / classification*,  genetics,  isolation & purification
Rotavirus Infections / diagnosis*,  veterinary*,  virology
Sensitivity and Specificity
Swine
Swine Diseases / diagnosis*,  virology
Virology / methods*
Chemical
Reg. No./Substance:
0/Antigens, Viral; 0/Capsid Proteins; 0/DNA Primers; 0/VP6 protein, Rotavirus

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