巢式聚合酶链反应检测A组轮状病毒基因型的研究

目的用巢式聚合酶链反应(PCR)对A组轮状病毒常见G、P基因型进行检测。方法根据A组轮状病毒不同型别共有的VP4、VP7基因设计的引物分别用于G、P分型的第1次PCR扩增,然后用可以区分不同G型和P型的A组轮状病毒的引物进行第2次PCR扩增。对不同型别A组轮状病毒标准毒株和68例A组轮状病毒腹泻的粪便标本进行了G、P型别的检测。结果不同型别A组轮状病毒标准毒株分别扩增出不同长度片段的条带。68例被检测的A组轮状病毒腹泻粪便标本中G分型以G3为主,P分型以P8为主,其余型别少见。结论巢式PCR检测A组轮状病毒基因型的分型具有较高的灵敏度,操作简便快速,适用于临床检测。

Detection of genotype of group A rotavirus by nested-polymerase chain reaction

Objective To develop a nested-polymerase chain reaction(PCR) to identify G and P genotypes of group A rotavirus. Methods Primers designed based on the common VP4 and VP7 gene sequence of group A rotavirus were utilized for the first run of PCR.The other primers that could identify group A rotavirus with G or P genotypes were adopted for the second run of PCR.The standard group A rotavirus with different genotypes and the G,P genotype of 70 stool specimens from patients with group A rotavirus diarrhea were detected. Results Different sizes of PCR products were amplified for standard group A rotavirus.G3 and P8 were the predominant genotypes in 70 stool specimens from patients with group A rotavirus diarrhea and other minor genotypes were also found. Conclusions Nested-PCR is sensitive for the detection of group A rotavirus genotypes.It is simple,rapid and particularly useful for clinic detection.