摘要
目的 建立一种灵敏、特异的 RT- PCR方法检测环境中的 HAV。方法 对两种 RT- PCR,即常规两步法 RT-PCR和一步法 RT- PCR检测 HAV的效果进行了观察 ,并采用半套式 PCR对扩增产物的特异性进行验证。结果 两种RT- PCR与半套式 PCR检测 HAV均得到预期条带 ,所设计的引物对其它的肠道病毒扩增呈阴性 ;在同等反应条件下 ,一步 RT- PCR的反应效率远高于常规 RT- PCR,二者的检测灵敏度也有显著性差异 ,一步 RT- PCR的检测灵敏度为 10TCID5 0 / ml,两步法 RT- PCR检测灵敏度为 10 3TCID5 0 / ml。结论 一步 RT- PCR较常规 RT- PCR易于操作 ,重复性好 ,不易污染 ,具有更高的反应效率和灵敏度。
Objective A sensitive and specific RT PCR approach was expected to detect HAV in environment Methods In the experiment,the conventional two step RT PCR was compared with the one step RT PCR and the semi nested PCR was involved to test the specificity of the amplified products Results The expected amplified products were achieved when HAV was detected by the two RT PCR approaches and the semi nested PCR,however,other enteroviruses were undetectable when detected by the two RT PCR approaches Under a same reaction condition,the reaction efficiency of one step RT PCR was significantly higher than that of conventional RT PCR,and the sensitivity showed significant differences between two methods The sensitivities of the one step RT PCR and the two step RT PCR were 10 and 10 3 TCID 50 /ml respectively Conclusion The results indicated that the one step RT PCR approach was easier to operate and repeat,less likely to be contaminated,and had higher reaction efficiency and sensitivity than those of the conventional RT PCR
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2001年第1期35-37,共3页
Journal of Environment and Health
基金
国家自然科学基金!资助 (39870 669)
