荧光定量PCR和酶联免疫吸附技术在手足口病肠道病毒快速检测中的应用比较

Application Comparison on Hand-Foot-Mouth Disease Rapid Detection of Enterovirus by Fluorescent Quantitative PCR and Enzyme-Linked Immunosorbent Assay

目的：比较荧光定量PCR（FQ-PCR）和酶联免疫吸附技术（ELISA）在手足口病肠道病毒快速检测中的应用价值。方法：选取疑似手足口病患儿200例,采集咽拭子、粪便、疱疹液标本232份,进行FQ-PCR检测,同时采用ELISA进行平行检测,比较两者的阳性检出率、灵敏度与特异性以及检测窗口期。结果：应用FQ-PCR方法,咽拭子、粪便、疱疹液中EV71、CoxA16、通用型肠道病毒核酸RNA的总阳性检出率分别为84.83%、81.82%、85.71%,差异无统计学意义（F=0.79,P〉0.05）。FQ-PCR方法咽拭子、粪便、疱疹液阳性检出率均高于ELISA法对血液的阳性检出率（χ2=3.17~5.11,P〈0.05）。FQ-PCR、ELISA诊断灵敏度分别为97.03%、87.50%（χ2=3.98,P〈0.05）,诊断特异性分别为97.22%、88.68%（χ2=3.85,P〈0.05）。在EV71、CoxA16及通用型肠道病毒方面,FQ-PCR的检测窗口期均早于ELISA,差异有统计学意义（P〈0.05）。结论：FQ-PCR在手足口病肠道病毒快速检测方面优于ELISA,具有更高的阳性检出率、灵敏度与特异性,以及更早的检测窗口期,对手足口病的早期诊断具有较高的临床指导意义和推广应用价值。

Objective： To compare the application on HFMD rapid detection of enterovirus by fluorescent quantitative PCR （FQ-PCR） and enzyme-linked immunosorbent assay （ELISA）. Methods： Two hundreds cases with suspected HFMD were selected, 232 samples were collected from throat swab, stool and herpes fluid, and were detected by FQ-PCR. At the same time, ELISA method was used to detect paraflelly, and the positive rate, sensitivity, specificity and detection window period were compared of two groups. Results： Total positive rate of EV71, CoxA16, universal enterovirus RNA were 84.83% , 81.82% and 85.71% by FQ-PCR method from throat swab, stool and herpes fluid, and there were no statistical difference among them （F = 0.79, P〉0.05 ）. Positive rates from throat swab, stool and herpes fluid detected by FQ-PCR were all higher than that of blood sample detected by ELISA （X2 = 3. 17 -5. 11, P〈0.05 ）. Diagnostic sensitivity of FQ-PCR and ELISA were 97.03% and 87.50% respectively （X2= 3.98, P〈0.05）, and diagnostic specificity was 97.22% and 88.68% （X2 =3.85, P〈0.05）. Detection window period of FQ-PCR on EV71, CoxAl6 and universal enterovirus were all earlier than that of ELISA （ P〈0.05 ）. Conclusion： FQ-PCR is better than ELISA on HFMD rapid detection of enterovirus, and it has higher positive rate, sensitivity and specificity, and earlier detection window period, so it has high clinical significance and application value on HFMD children＇ s early diagnosis.