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Journal of Clinical Microbiology, October 2008, p. 3330-3337, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00432-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Determination of Human Rotavirus VP6 Genogroups I and II by Reverse Transcription-PCR{triangledown} ,{dagger}

Yi-Pei Lin,1 Chuan-Liang Kao,1,2 Sui-Yuan Chang,1,2 Koki Taniguchi,3 Pei-Yung Hung,1 Hsueh-Ching Lin,1 Li-Min Huang,4 Hsueh-Hung Huang,2 Jyh-Yuan Yang,5 and Chun-Nan Lee1,2*

Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University,1 Department of Laboratory Medicine,2 Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan, Republic of China,4 Department of Virology and Parasitology, Fujita Health University School of Medicine, Toyoake, Aichi, Japan,3 Center for Disease Control, Department of Health, The Executive Yuan, Taipei, Taiwan, Republic of China5

Received 4 March 2008/ Returned for modification 22 April 2008/ Accepted 22 July 2008

Based on nucleotide sequence and phylogenetic analysis of the partial VP6 genes, group A rotaviruses can be mainly differentiated into two genogroups. In this study, a method employing reverse transcription-PCR (RT-PCR) and degenerate primers was established to assign the VP6 genogroup. VP6 genogroup I and genogroup II could be determined according to the sizes of the amplicons: 380 and 780 bp, respectively. The VP6 genogroup of human reference strains of G1 to G4 and G9 types and RotaTeq vaccine strains could be properly assigned by RT-PCR. Eighty rotavirus-positive fecal samples were subjected to enzyme-linked immunosorbent assay (ELISA), RT-PCR, and sequencing of the partial VP6 gene for subgroup and genogroup determination. The results correlated well among these three methods, except for seven samples whose subgroups could not be determined by ELISA. VP6 genogroups of another 150 rotavirus strains recovered between 1981 and 2005 were determined by RT-PCR and sequencing, and the same results were obtained by these two methods. Furthermore, an additional 524 rotavirus-positive fecal samples were tested by RT-PCR, and the VP6 genogroups could be easily determined. The RT-PCR assay developed here provided a reliable and convenient method for assigning the VP6 genogroups of human rotaviruses with a wide range of genetic variation.

* Corresponding author. Mailing address: Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, No. 1, Chang-Te St., Taipei 100, Taiwan. Phone: 886-2-23123456 ext 66905. Fax: 886-2-23711574. E-mail: cnalee{at}ntu.edu.tw

{triangledown} Published ahead of print on 30 July 2008.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, October 2008, p. 3330-3337, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00432-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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