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目的 观察SARS患者各种临床标本的带毒状况 ,分析SARS冠状病毒 (SARS CoV)在患者体内的动态变化及其临床意义。方法 设计针对SARS CoVP区和N区的 2套引物 ,应用巢式RT PCR对 84例SARS患者的血、尿、痰、粪便、咽拭子等 4 94份不同发病时间收集的标本进行病毒定性检测 ,并将PCR产物直接进行DNA序列分析。结果 两套引物均能较好地扩增出部分标本SARS CoV基因的特异性片段。在所有标本中痰液阳性检出率最高 ,联合检测多种标本或同时应用两套引物进行PCR扩增可明显提高标本阳性检出率。跟踪观察发现 ,SARS患者发病第 2周时PCR阳性检出率最高 ,第 3～ 4周次之 ,但各种标本阳性检出率的动态变化不尽相同。结论 应用巢式RT PCR进行SARS早期诊断和检测是一种可行的方法 ,针对SARS CoV基因组不同区域设计多对引物 ,联合检测多种临床标本可明显提高阳性检出率 ;研究SARS CoV的检出变化特点将为SARS的临床治疗提供一定的依据
Objective To observe the virulence status of SARS patients with various clinical specimens and to analyze the dynamic changes and clinical significance of SARS coronavirus (SARS CoV) in patients. Methods Two sets of primers were designed for CoVV and N regions of SARS. Nested reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the virus in 84 samples collected from 94 SARS patients at different times of onset, including blood, urine, sputum, stool and throat swabs , And PCR products were directly DNA sequence analysis. Results Both primers could amplify the specific fragment of SARS CoV gene in some samples. In all specimens sputum positive detection rate of the highest combined detection of a variety of specimens or two sets of primers for PCR amplification can significantly increase the positive detection rate of specimens. Follow-up observation found that the highest detection rate of PCR positive cases occurred in the second week of SARS patients, ranging from 3 to 4 weeks, but the dynamic changes of the positive detection rates of various specimens were different. Conclusion It is feasible to use Nested RT-PCR to diagnose and detect SARS in early stage. To design multiple pairs of primers for different regions of SARS CoV genome, the combined detection of various clinical specimens can significantly improve the positive detection rate. The detection of SARS CoV The characteristics of the changes will provide some evidences for the clinical treatment of SARS