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目的为严重急性呼吸综合征(SARS)病例的诊断提供科学依据。方法连续、系统地采集2003年12月至2004年1月广东省新发4例SARS怀疑病例不同发病时间的各类标本,采用酶联免疫吸附测定(ELISA)、免疫荧光测定(IFA)、中和试验等血清学检测技术和逆转录聚合酶链反应(RT-PCR)、荧光定量聚合酶链反应等分子生物学技术检测采集的标本,并对检测结果进行分析。结果血清学检测结果显示,4例病例在发病后6~8d,均可检测到SARS-CoVIgM和IgG抗体;前3例病例SARS-CoV抗体有4倍或4倍以上增长,第4例病例SARS-CoVIgG抗体虽无4倍增长,但却有明显的抗体阳转过程。采用ELISA、IFA和中和试验的检测结果基本一致。用荧光定量PCR检测咽拭等标本的SARS-CoV核酸,结果只有病例1的3份咽拭标本阳性,其他标本均阴性。对阳性标本,采用RT-PCR扩增出SARS-CoV的M、N、S基因并测序,S基因序列和种系发生分析提示,该病毒与从广东省果子狸中分离到的SARS毒株最接近。未能从这4例SARS病例的任何标本中分离到病毒。结论检测结果证明,该4例病例报告可以确认为SARS实验室确诊病例。
Objective To provide a scientific basis for the diagnosis of severe acute respiratory syndrome (SARS) cases. Methods A total of 4 newly diagnosed cases of SARS in Guangdong Province from December 2003 to January 2004 were collected. ELISA, immunofluorescence (IFA) And test serological detection techniques and reverse transcription polymerase chain reaction (RT-PCR), fluorescence quantitative polymerase chain reaction and other molecular biology techniques to detect the collected specimens, and the test results were analyzed. Results The results of serological test showed that SARS-CoVIgM and IgG antibodies were detected in all the four cases from 6 to 8 days after the onset of disease. The first 3 cases showed a 4-fold or 4-fold increase in SARS-CoV antibodies and the fourth case SARS Although there was no 4-fold increase in the -CoVIgG antibody, there was a clear positive anti-body rotation process. Using ELISA, IFA and neutralization test results are basically the same. Fluorescent quantitative PCR was used to detect the SARS-CoV nucleic acid in pharyngeal swab specimens. Only 3 swab swab specimens in case 1 were positive, and all other specimens were negative. The positive samples were amplified by RT-PCR from SARS-CoV M, N, S gene and sequencing, S gene sequence and phylogenetic analysis suggest that the virus and Guangdong Province civets from SARS strains most Close to The virus was not isolated from any of the 4 SARS cases. Conclusion The test results show that the 4 cases can be confirmed as SARS laboratory confirmed cases.