Nigerian rotavirus serotype G8 could not be typedby PCR due to nucleotide mutation at the 3′ endof the primer binding site
Nigerian rotavirus serotype G8 could not be typedby PCR due to nucleotide mutation at the 3′ endof the primer binding site
复制标题
尼日利亚轮状病毒G8血清型因引物结合位点3′端核苷酸突变而无法通过PCR分型
作者：
M. Adah;A. Rohwedder;O. D. Olaleyle;H. Werchau
从尼日利亚分离的一株轮状病毒HMG 89株，经PCR鉴定为G3型。对来自PCR产物的cDNA克隆进行测序，该cDNA克隆在北方印迹中与同源尼日利亚轮状病毒株HMG 89和实验室参考株69 M的RNA片段9杂交，但不与其他哺乳动物A组轮状病毒杂交。VP 7基因9序列长1060个核苷酸，在1034-1035位有两个碱基缺失。对先前PCR血清分型试验中使用的引物（aAT 8）进行的序列分析显示，引物结合位点3′端的三个核苷酸碱基之一发生突变，这是我们无法对样本中的G8菌株进行血清分型的原因。这些发现表明，PCR分析可以，虽然很少，导致错误的轮状病毒分型，由于在引物结合区的突变数量少。
SummaryA rotavirus strain HMG89 from Nigeria with short electrophoretic pattern was typed G3 by PCR. A cDNA clone from the PCR product which hybridised in Northern blots to RNA segment 9 of the homologous Nigerian rotavirus strain HMG89 and laboratory reference strain 69M but not to other mammalian group A rotaviruses was sequenced. The VP7 gene 9 sequence is 1060 nucleotides long with two base deletions at positions 1034–1035. Sequence analysis of the primer (aAT8) used in the previous PCR serotyping assay revealed a mutation in one of the three nucleotide bases at the 3′ end of the primer binding site accounting for our inability to serotype G8 strains in our samples. These findings demonstrate that PCR analysis can, albeit infrequently, lead to error in typing of rotaviruses due to small numbers of mutations in the primer binding region.