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为明确严重急性呼吸综合症(SARS)冠状病毒N蛋白在SARS实验室早期诊断中的作用,通过微量中和试验及酶联免疫方法、间接免疫荧光法检测疑似病人恢复期血清(大于28天)中SARS IgG抗体,确诊SARS患者。同时收集发病不同时期SARS及普通发热病人血清,利用酶联免疫方法检测SARS CoVN蛋白,并与荧光定量PCR早期诊断方法相比较。共检测:广州地区2003年12月～2004年1月新发4例确诊SARS患者不同时期的血液和咽漱液标本;恢复期血清SARS CoV中和抗体阳性病人不同时期的血清46份;广州地区2003年1月～4月临床确诊SARS患者159例的血清和56例疑似患者血清;非SARS普通发热病人血清97份;正常人体检血清100份。结果:4例新发SARS患者的不同时期标本中,3例患者急性期血均检出N蛋白,优于常用的荧光定量PCR检测方法。46份SARS CoV中和抗体阳性的血清标本,N蛋白检出率为100%。159例临床确诊病例中,发病早期5天以内SARS CoVN蛋白的检出率为92 3%,随后呈现逐步下降的趋势,在发病第18天仍可检出。56例临床疑似患者发病早期也有23 2%检出率。而97例普通发热病人及100份正常人血清中均未能检测出SARS CoVN蛋白。表明在血清中检测SARS冠状病毒N蛋白的方法敏感性和特异性都好,对SARS实验室早期诊断具有重要作用。
To clarify the role of severe acute respiratory syndrome (SARS) coronavirus N protein in the early diagnosis of SARS laboratory, the convalescent sera of suspected patients (more than 28 days) were detected by indirect immunofluorescence with micro-neutralization assay and enzyme-linked immunosorbent assay SARS IgG antibody, diagnosed SARS patients. At the same time, the serum of SARS patients and common fever patients were collected at the same time, and the protein of SARS CoVN was detected by enzyme-linked immunosorbent assay (ELISA), and compared with the method of early detection of fluorescence quantitative PCR. A total of 4 samples of blood and pharyngeal rhesus were collected from 4 cases of newly diagnosed SARS patients in Guangzhou area from December 2003 to January 2004. 46 cases of serum from different phases of SARS CoV neutralizing antibody were detected in Guangzhou area Serum of 159 SARS patients and 56 suspected patients with clinically confirmed SARS in January 2003 to April 2003; 97 serums of non-SARS common fever patients; 100 serum samples of normal people. Results: Among the four cases of newly diagnosed SARS patients, N protein was detected in all acute stage blood samples, which was better than the common fluorescence quantitative PCR method. Forty-six SARS-CoV neutralizing antibody-positive serum samples were detected with 100% N protein. Among the 159 clinically confirmed cases, the detection rate of CoVN protein in SARS within 5 days of onset was 92.3%, then showed a gradual downward trend, which was still detectable on the 18th day of onset. 56 cases of clinical suspicion of early onset also 23 2% detection rate. SARS CoVN protein was undetectable in the serum of 97 patients with normal fever and 100 normal persons. The results showed that the method of detecting SARS-CoV N protein in serum was sensitive and specific, which played an important role in the early diagnosis of SARS.