EB病毒感染实验室诊断方法探讨

Analysis of laboratory assays applied for diagnosis of Epstein-Barr virus

目的探讨EB病毒(EBV)感染的实验室诊断方法。方法设计针对EBV基因组的引物和荧光标记探针,利用实时荧光定量聚合酶链反应(FQ-PCR)检测70例外周血标本。其中17例是确诊EBV感染及传染性单核细胞增多症患儿,27例是临床高度怀疑EBV感染患儿,26例是临床非EBV感染的患儿,并与检测EBV抗体VCA-IgM的酶联免疫吸附试验(ELISA)进行比较。结果确诊组、疑似组和对照组患儿FQ-PCR检测EBV的阳性率分别为100.00%、81.48%和0.00%。与此对应VCA-IgM的阳性率分别为100.00%、25.93%和0.00%。确诊组EBV DNA的平均拷贝数为1.15×105/m l,疑似组为4.38×104/m l,均明显高于对照组(P<0.05)。结论与VCA-IgM检测方法相比,FQ-PCR检测到EBV感染的阳性率更高,EBV DNA的定量检测可帮助诊断儿科活动性EBV感染,尤其是可疑患儿的临床诊断。

Objective To analyze laboratory assays for diagnosis of Epstein-Barr virus（EBV）. Methods Primers and probes were designed to identify genome of EBV and 70 samples were analyzed by fluorescence quantitative polymerase chain reaction （FQ-PCR）. These samples include 17 children with diagnosed EBV infection and（or） infectious mononucleosis,27 with highly suspected EBV infection and 26 without EBV infection. The results were compared with that of EBV VCA-IgM detected by enZyme-linked immunosorbent assay （ELISA）. Results Positive rates of FQ-PCR for three groups were 100.00% ,81.48% and 0.00% respectively and positive rates of VCA-IgM were 100.00%, 25.93% and O. 00% respectively. The mean virus DNA copies of diagnosed group were significantly higher than that of negative control group （P 〈 0.05 ）, which were 1.15 × 10^5/ml, 4.38 × 10^4/ml respectively. Conclusions Positive rate of EBV infection determined by the assay of FQ-PCR is higher than that by EBV VCA-IgM. Quantification of EBV DNA could be helpful for diagnosis of clinical EBV infection especially for those patients with suspected EBV infection.