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

作者：Thongprachum Aksara; Chaimongkol Natthawan; Khamrin Pattara; Pantip Chansom; Mizuguchi Masashi; Ushijima Hiroshi; Maneekarn Niwat^*

来源：Journal of Virological Methods, 2010, 168(1-2): 191-196.

DOI：10.1016/j.jviromet.2010.05.013

• Human rotavirus
• Porcine rotavirus
• Subgroup
• Genogroup
• Multiplex RT-PCR
• Monoplex RT-PCR

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.

• 出版日期2010-9