流行性感冒病毒应急检测中缩减荧光定量逆转录-聚合酶链反应时间的实验研究  

Study on Time Decrease Using Real-time RT-PCR in Emergency Detection of Influenza

作　　者：葛琼[1] 龚黎明[1] 卢亦愚[1] 严菊英[1] 茅海燕[1] 周敏[1] 李敏红[1] 

机构地区：[1]浙江省疾病预防控制中心,杭州310009

出　　处：《中国计划免疫》2007年第2期158-160,共3页Chinese Journal of Vaccines and Immunization

摘　　要：目的在相同的敏感度与特异性条件下,进一步缩减流行性感冒(流感)病毒荧光定量逆转录-聚合酶链反应(RT-PCR)的时间,为应急检测服务。方法缩短荧光定量RT-PCR经典检测方法中不同反应阶段的时间参数,并与经典方法进行比较,考察改良方法的检测敏感性与特异性。结果用经典方法与缩短某些时间参数的改良方法检测甲3型流感病毒,结果其特异性与灵敏度差异无显著的统计学意义,并可将检测时间缩短1h左右。结论适当缩短经典荧光定量RT-PCR方法中的逆转录、变性以及延伸时间后,灵敏度和特异性无显著影响,这对于突发疫情的应急检测具有实用价值。Objective To evaluate the feasibility of shortening the reaction time of real-time RT-PCR for influenza virus to improve the diagnostic efficiency in emergent infectious epidemics. Methods The time required in each step of the original real-time RT-PCR method was shortened. The sensitivity and specificity of this improved real-time RT-PCR method was evaluated by comparing with the original method. Results The sensitivity and specificity had no significant difference between the improved and original real-time RT-PCR method to detect influenza virus A3. Furthermore the time required to complete the real-time RT-PCR method was shortened about one hour by the improved method. Conclusions The sensitivity and specificity were not affected by properly shortening the reactive time for reverse transcription, denaturation and extension steps of the real-time PCR. These results were very valuable for diagnosis of the emergent infectious disease.

关 键 词：荧光定量逆转录-聚合酶链反应 缩短反应时间 敏感性 

分 类 号：R373.13[医药卫生—病原生物学]