四种商品化诺如病毒实时荧光RT-PCR检测试剂盒的效果比较

Comparison of four commercial real-time RT-PCR kits for norovirus detection

目的比较四种商品化诺如病毒实时荧光RT-PCR检测试剂盒的效果。方法将美国疾控中心诺如病毒检测方法作为标准方法,使用4种商品化试剂盒对30份已知诺如病毒基因型阳性样本(5种GI和8种GII基因型)和50份腹泻患者粪便样本进行检测,对检测结果进行比较。同时对轮状病毒、星状病毒、肠道腺病毒和札如病毒进行特异性检测。结果30份已知基因型样本四种商品化试剂盒与标准方法均检测阳性。轮状病毒、星状病毒、肠道腺病毒和札如病毒检测阴性。试剂盒A、B、C、D和标准方法分别检测50份腹泻样本,检出率分别为52.0%(26/50)、46.0%(23/50)、50.0%(25/50)、48.0%(24/50)、54.0%(27/50);试剂盒A、B、C、D与标准方法的一致性分别为96.3%、85.2%、92.6%和88.9%。试剂盒C的GI检测范围为10^4~10^8 copies/μL,GII检测范围为10^2~10^8 copies/μL。结论四种试剂盒均能检测本研究出现的诺如病毒基因型,试剂盒A和C检测效果优于试剂盒B和D。

Objective To compare the effectiveness of four commercial norovirus real-time RT-PCR detection kits.Methods Using the norovirus detection method from US Centers for Disease Control and Prevention as the standard method,30 specimens of known norovirus genotypes(5 GI and 8 GII genotypes)and 50 stool specimens from patients with diarrhea were detected by four commercial norovirus real-time RT-PCR detection kits to compare the effectiveness of the kits.Detections for astrovirus,enteric adenovirus and sapovirus were performed to determine the specificity of these kits.Results All 30 specimens of known genotypes were detected as positive by four commercial kits and the standard method.Rotavirus,astrovirus,enteric adenovirus and sapovirus were detected negative by four kits.The detection rate of four kits(A,B,C and D)and standard method for 50 diarrhea samples were 52.0%(26/50),46.0%(23/50),50.0%(25/50),48.0%(24/50)and 54.0%(27/50),respectively.The consistency rates of four kits(A,B,C and D)with the standard method were 96.3%,85.2%,92.6%and 88.9%,respectively.For the kit C,the detection ranges of GI and GII were 10^4-10^8 copies/μL and 10^2-10^8 copies/μL,respectively.Conclusions All genotypes of norovirus in this study were detected by the four kits,and the detection abilities of the kit A and C were better than kit B and D.