title: A novel multiplex RT-PCR for identification of VP6 subgroups of human and porcine rotaviruses.
abstract: 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.affiliation: Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
authors.forename: Aksara
authors.initials: A
authors.lastname: Thongprachum
authors.forename: Natthawan
authors.initials: N
authors.lastname: Chaimongkol
authors.forename: Pattara
authors.initials: P
authors.lastname: Khamrin
authors.forename: Chansom
authors.initials: C
authors.lastname: Pantip
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizuguchi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ushijima
authors.forename: Niwat
authors.initials: N
authors.lastname: Maneekarn
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D036022:Capsid Proteins / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology; D006801:Humans; D017720:Molecular Epidemiology / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012401:Rotavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D012400:Rotavirus Infections / Q000175:diagnosis* / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D014773:Virology / Q000379:methods*
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; C061332:VP6 protein, Rotavirus