深圳市传染性非典型肺炎患者聚合酶链反应及血清学检测研究

Studies on PCR and serological detection of SARS in Shenzhen

目的 探讨传染性非典型肺炎 (SARS)的逆转录聚合酶链反应 (RT PCR)及血清学检测方法。方法 收集SARS临床诊断病例咽漱液标本 2 3份和血清 10 8份 ,应用SARS冠状病毒分子信标RT PCR、间接免疫荧光试验 (IFA)和酶联免疫吸附试验 (ELISA)分别进行了病原学和血清学检测。结果 2 3份SARS患者咽漱液用分子信标RT PCR检测 ,阳性 10份 ,阳性率为 4 3 4 8% (10 /2 3)。IFA法检测血清中SARS冠状病毒IgM抗体阳性率 (17 78% )明显低于ELISA法 (6 8 89% ) ,两者IgM阳性率存在显著性差异 (χ2 =2 3 94 ,P <0 0 0 5 ) ,两者IgG抗体阳性率无显著性差异 (χ2 =0 78,P >0 0 5 )。结论 IFA用于SARS病毒IgM检测 ,敏感性比ELISA差。IgM抗体检测宜于发病 10d后取标本 ;IgG抗体检测适于发病 2周后取标本。分子信标RT PCR可发展成为一种快速的SARS诊断检测方法 ,抗体检测简便、快速 ,可作为临床辅助诊断指标。

Objective To explore the RT-PCR and serological detection methods for SARS. Methods 23 throat washes, 108 sera of SARS patients were assayed by RT-PCR with molecular beacon, IFA and ELISA, respectively. Results The positive rate of RT-PCR was 43.48%(10/23)by molecular beacon method. The positive rate(17.78%)of specific IgM against SARS virus by IFA was lower than that by ELISA(68.89%). The difference was significant (χ 2=23.94, P <0.005). The positive rate of IgG detected by IFA was similar to that by ELISA (χ 2=0.78, P >0.05). Conclusion The sensitivity of IFA for SARS IgM detection was lower than that of ELISA. Sampling date should be 10 days after onset of the disease for IgM detection and 14 days for IgG detection. Both ELISA and IFA are rapid and simple. They could be used to assist diagnosis of SARS coronavirus infection. RT-PCR with molecular beacon may become a rapid diagnostic method for SARS detection.