A novel multiplex RT-PCR for identification of VP6 subgroups of human and porcine rotaviruses

• Main Authors: Thongprachum A., Chaimongkol N., Khamrin P., Pantip C., Mizuguchi M., Ushijima H., Maneekarn N.
• Format: Article
• Language: English
• Published: 2014
• Online Access: http://www.scopus.com/inward/record.url?eid=2-s2.0-77955097047&partnerID=40&md5=a0f943a31409771260739aef98c8daf5; http://www.ncbi.nlm.nih.gov/pubmed/20546787; http://cmuir.cmu.ac.th/handle/6653943832/2548
• Institution: Chiang Mai University

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. © 2010 Elsevier B.V.