3种方法对发热伴血小板减少综合征检测结果比较

目的 采用实时荧光PCR法、酶联免疫吸附法及细胞培养分离病毒株的方法,对2013年临床疑似发热伴血小板减少综合征患者急性期血进行检测比较,寻求快速、准确的鉴定新布尼亚病毒感染的方法。方法 对180例临床疑似病例的急性期血液首先采用实时荧光PCR法检测,筛选出新布尼亚病毒核酸阳性的样本,同时将阳性样本采用酶联免疫吸附的方法检测抗体及采用VERO-E6细胞培养的方法分离病毒株。结果 180份样本经实时荧光PCR法检测,阳性率为42.78%(77/180 Ct值≤35曲线形态与阳性对照一致呈S型的),ELISA方法检测抗体阳性率为44.16%(34/77),将核酸检测阳性的77份样本全部感染VERO-E6细胞获得病毒株32株,阳性率为38.55%(32/77)。结论 实时荧光PCR法更适合于新布尼亚病毒感染者早期实验室快速诊断,发病在1w左右的病例不适合用ELISA的方法做确证实验。细胞培养分离病毒虽然是金标准,但耗时、费力,对早期诊断意义不大,但适于研究或疫苗研发。

Three methods for detecting severe fever with thrombocytopenia syndrome bunyavirus

In this study,we detected the blood of clinical patients with suspected acute fever with thrombocytopenia syndrome （SFTSV） by three methods including real-time PCR,enzyme-linked immunity assay and virus strains isolation in cell culture,in order to find the most appropriate one for the identification of Bunia virus infection quickly and accurately.Firstly,we detected 180 cases of acute blood which were clinically suspected by real-time PCR,and screened out positive samples of nucleic acid of SFTSV.At the same time,we detected the antibodies of positive samples by ELISA and isolated strains by VERO-E6 cell culture.The positive rate of 180 samples was 42.78％ detected by PCR assay,（77/180,Ct value≤35 curve shape was consistent with the positive one which showed a S-type）,the positive rate of the antibody was 44.16％ by detection of ELISA （34/77）,the 77 samples which were positive in nucleic acid test were infected VERO-E6 cells to obtain 32 samples of virus strain,and the positive rate was 38.55％ （32/77）.Results suggested that real-time PCR detection was more suitable for the early rapid laboratory diagnosis of new Bunia virus infection with high isolation rate.In the blood,the virus could maintain a long time,and the IgM antibodies emerged later.It is not suitable for ELISA detection to conduct confirmatory test when the cases have been breaking out for a week.Cases were mainly occurred between July and September,and the age of the cases were mainly 40 to 60 years-old,associating with the flow of the tick breeding season.