严重急性呼吸综合征病原体的检测方法探讨

Study on the methods for detection of SARS-associated with Coronavirus

目的 在实验室建立SARS病原学检测方法。方法 采集临床诊断为SARS患者、疑似病人漱口液、用细胞培养分离,荧光定量PCR方法检测。病原体用间接免疫荧光鉴定。结果 漱口液、咽拭子液样本57份作病原体分离,阳性数5份,阳性率8.8％尸解标本7份,其中肺部组织3份、气管黏液、淋巴液、肺积液、心包液各1份。结果细胞分离到7株病原体。阳性率100％。荧光定量PCR方法检测临床确诊SARS患者标本57份,检出阳性结果38份,阳性率为66.7％。密切接触者标本14份,检出阳性结果7份,阳性率为50.0％。疑似患者标本59份中检出阳性12份,阳性率为20.3％。两种方法的比较,统计学上有显著性差异(P<0.001)。结论用荧光定量PCR方法检测SARS患者病原体敏感性高,可靠性和重复性好,优于细胞病毒分离的方法,可以作为SARS临床早期诊断和流行病学调查的重要指标之一。

To develop and compare several methods for detection of SARS-associated with Coronavirus, the specimens from identified patients with SARS, suspect patients and autopsy were collected respectively. Methods including cell culture with three different cell, fluorescent quantitative polymerase chain reaction, and indirect immunofluorescent assay were applied. The results showed that in 57 specimens detected by cell culture method, 5 from throat-swab and throat-washing showed positive, and 7 specimens from autopsy showed positive. In 130 specimens detected by fluorescent quantitative polymearase chain reaction method, 38 in 57 specimens from cases of identified SARS patients, 7 in 14 specimens from the case of closely contact, and 12 in 59 specimens from the cases of suspect SARS patients showed positive. It concludes that the development of rapid, accurate and efficient methods is important for epidemical investigation and clinical diagnosis of SARS.