https://scholars.lib.ntu.edu.tw/handle/123456789/503562
標題: | Determination of human rotavirus VP6 genogroups I and II by reverse transcription-PCR | 作者: | Lin Y.-P. CHUAN-LIANG KAO SUI-YUAN CHANG Taniguchi K. Hung P.-Y. Lin H.-C. LI-MIN HUANG Huang H.-H. Yang J.-Y. Lee C.-N. |
公開日期: | 2008 | 卷: | 46 | 期: | 10 | 起(迄)頁: | 3330-3337 | 來源出版物: | Journal of Clinical Microbiology | 摘要: | 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. Copyright ? 2008, American Society for Microbiology. All Rights Reserved. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/503562 | ISSN: | 0095-1137 | DOI: | 10.1128/JCM.00432-08 | SDG/關鍵字: | protein VP4; protein VP6; protein VP7; Rotavirus nonstructural protein 4; Rotavirus vaccine; amplicon; article; controlled study; enzyme linked immunosorbent assay; feces analysis; female; genotype; human; Human rotavirus; major clinical study; male; nonhuman; nucleotide sequence; phylogeny; priority journal; reverse transcription polymerase chain reaction; Antigens, Viral; Capsid Proteins; Feces; Genotype; Glycoproteins; Humans; Molecular Sequence Data; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral; Rotavirus; Rotavirus Infections; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Toxins, Biological; Viral Nonstructural Proteins; Human rotavirus A; Rotavirus; Rotavirus A |
顯示於: | 醫學檢驗暨生物技術學系 |
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