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以往甲型肝炎病毒（ＨＡＶ）的检测主要依赖细胞培养法，但此法所需时间长、敏感性低、特异性不强。虽然反转录ＰＣＲ等技术已被用于粪便或食品中ＨＡＶ的检测，但常规反转录ＰＣＲ操作繁琐、污染环节多，极易造成污染，致使实验结果不稳定或出现假阳性结果。为克服ＰＣＲ检测ＨＡＶ的缺点和不足，我们在实验基础上建立了一步单管反转录ＰＣＲ技术检测水中的ＨＡＶ，采用热裂解法提取样品中ＨＡＶ病毒的总ＲＮＡ，反转录与ＰＣＲ反应一步进行，整个反应在一个反应管中进行，操作方法较简单，中间可能污染的环节较少，因此，最大限度的减少了由环境造成的可能污染。我们设计合成的引物（Ｈ１—Ｈ２）在ＨＡＶ病毒核酸的ＶＰ１区，只特异地扩增ＨＡＶ核酸片断，具有较强的特异性。改进后的反转录ＰＣＲ的敏感性与常规反转录ＰＣＲ比有所提高，可检测到细胞培养悬液或水中１０个ＴＣＩＤ＿（５０）的ＨＡＶ。
In the past, detection of Hepatitis A virus (HAV) mainly relied on cell culture, but this method requires long time, low sensitivity and low specificity. Although techniques such as reverse transcription PCR have been used for the detection of HAV in faeces or foodstuffs, routine reverse-transcription PCR operations are cumbersome and involve many pollution steps, which can easily lead to contamination, resulting in unstable experimental results or false-positive results. In order to overcome the shortcomings and deficiencies of HAV detection by PCR, we established a one-step reverse transcriptase-PCR assay to detect HAV in water. The total RNA of HAV virus was extracted by thermal cleavage. The reverse transcription and PCR reaction , The entire reaction is carried out in a reaction tube, the operation method is relatively simple, there are few possible contamination steps in the middle, and therefore, the possible pollution caused by the environment is minimized. We designed the synthetic primers (H1-H2) in the VP1 region of the HAV virus nucleic acid, only specifically amplified HAV nucleic acid fragments, with strong specificity. The sensitivity of the improved reverse transcriptase PCR was improved compared with that of the conventional reverse transcriptase PCR, and 10 TCID50 HAVs could be detected in the cell culture suspension or water.