实时荧光定量RT-PCR检测急性出血性结膜炎CoxA24v病毒  

Rapid detection of CoxA24v from acute hemorrhagic conjunctivitis by real-time RT-PCR

作　　者：李洁[1] 黄芳[1] 郭婧[1] 崔淑娟[1] 贾蕾[1] 李锡太[1] 黎新宇[1] 王全意[1] 

机构地区：[1]北京市疾病预防控制中心传染病地方病控制所,北京100013

出　　处：《中国病原生物学杂志》2011年第6期411-413,423,共4页Journal of Pathogen Biology

摘　　要：目的对检测急性出血性结膜炎(AHC)病原CoxA24v的实时荧光定量RT-PCR方法进行特异性、敏感性、稳定性等方面的评估。方法利用实时荧光定量RT-PCR检测肠道病毒CoxA24v和其他几种肠道病毒,验证方法的特异性;根据TCID50进行倍比稀释,验证该方法的敏感性;将样品进行倍比稀释,做重复试验,验证方法的准确性。利用实时荧光定量RT-PCR对疑似AHC患者眼拭子进行检测。结果该方法对CoxA24v病毒的检测具有高度的特异性,对柯萨奇病毒A16、肠道病毒71等其他肠道病毒均无交叉反应。检测的灵敏度达0.1TCID50。用该方法对6个不同浓度的CoxA24 v进行5次检测,获得的结果重复性好。采用该方法检测疑似AHC患者标本,阳性者均测序,证明是Cox-A24v,未出现假阳性。结论实时荧光定量RT-PCR法是一种快速、敏感、特异、准确的方法,适用于急性出血性结膜炎的早期诊断和鉴别诊断。Objective To further investigate the specificity,sensitivity,and accuracy of a established method of real-time RT-PCR to detect coxsackievirus A24 variant（CoxA24v）,a cause of acute hemorrhagic conjunctivitis（AHC）.Methods The specificity of the method was evaluated by detecting CoxA24v and several other enteroviruses.The CoxA24v strain was diluted in accordance with its TCID50 and the sensitivity of real-time RT-PCR was evaluated.The experiment was repeated and accuracy of the assay was evaluated.Clinical specimens from suspected cases were subjected to this assay.Results Real-time RT-PCR detected CoxA24v with a high level of specificity,and there were no cross reactions with common enteroviruses such as CoxA16 and EV71.The sensitivity of this test was up to 0.1 TCID50.Six different concentrations of CoxA24v were detected five times using this method.The results were very accurate.Clinical specimens from suspected cases were analyzed using this method,and positive PCR products were all sequenced.Results indicated CoxA24v and produced no false positives.Conclusion A real-time PCR assay was a quick,sensitive,specific,and accurate method that is suited to early diagnosis and differential diagnosis of AHC.

关 键 词：实时荧光定量RT-PCR CoxA24v 急性出血性结膜炎 

分 类 号：R373.9[医药卫生—病原生物学]