广东地区SARS标本检测方法的对比研究  

Comparison and assessment of the methods for determination of SARS specimens in Guangdong Province

作　　者：黄平[1] 李晖[2] 陈秋霞[2] 郑焕英[2] 俞守义[1] 鄢心革[2] 陈清[1] 黄吉城[2] 

机构地区：[1]第一军医大学流行病学教研室,广东广州510515 [2]广东省疾病预防控制中心,广东广州510300

出　　处：《中国热带医学》2004年第6期900-902,共3页China Tropical Medicine

基　　金：国家自然科学基金资助 (30 340 0 1 4 )

摘　　要：目的　比较和评价严重急性呼吸综合征的检测方法。　方法　采用巢式RT -PCR和实时荧光定量PCR检测病人咽痰标本 ,采用ELISA方法检测血清IgG抗体 ,同时采用细胞培养技术对咽痰标本进行病毒分离 ,并对检测结果进行统计学分析。　结果　两种RT -PCR检测结果完全一致 ;巢式RT -PCR与荧光定量PCR检测比较 ,两种检测方法之间的吻合度较强 ,检测结果基本一致 ;比较PCR方法和ELISA方法检测病毒基因和IgG抗体 ,结果发现PCR方法与ELISA方法之间的吻合度很弱 ,两种方法对于临床诊断具有互补性 ;采用细胞培养 (Vero -E6细胞株 )方法 ,分离了 13株SARS -CoV毒株。　结论　采用巢式RT -PCR和实时荧光定量PCR、ELISAIgG抗体检测和细胞培养技术对于SARS临床诊断、研究、预防和控制具有重要意义 ,目前的诊断技术已稳定可靠 ;但确诊周期过长 ,同时须防止标本污染和实验室污染。Objective To Compare and assess the methods for detection of SARS. Methods The methods of RT-PCR and real-time PCR for detection of SARS-CoV RNA from the pharyngeal swabs and sputum and the methods of ELISA for detection of SARS-CoV IgG antibody in sera and the cell culture for isolation of SARS-CoV strains were used. BNI primer and CDC primer were used in RT-PCR. The results of detective experiments were analyzed. Results The results obtained by using the RT-CR and real-time PCR are consistent and strong coincidence was observed in the two methods. While the coincidence of PCR is weak as compared with that of ELISA in detection of viral gene and IgG antibody. But they are complimentary for clinical diagnosis. Thirteen SARS -CoV strains were isolated and differentiated by culturing Vero-cells. Conclusion It is of important diagnostic, preventive, research and control significance in detection of SARS virus by RT-PCR and real-time PCR and serum IgG antibody by ELISA with cell culture method. At present, the diagnostic technique are stable and reliable, but the cycle for confirmative diagnosis is too long in addition to the need of prevent from contamination of samples and laboratory contamination.

关 键 词：巢式RT-PCR SARS 痰标本 IgG抗体 吻合 标本检测 ELISA方法 PCR方法 毒株 RT-PCR检测 

分 类 号：R563.1[医药卫生—呼吸系统] R446[医药卫生—内科学]