RT-PCR法检测贝类中的甲肝病毒的研究

在世界范围内,甲肝病毒是与食用贝类有关的主要传染性疾病之一。由于贝类中含有PCR抑制剂以及病毒富集过程中病毒的回收率低,阻碍了天然污染的贝类中HAV的PCR检测。研究中建立了一种经苷氨酸缓冲液洗涤,2次PEG沉降富集病毒,然后进行RNA提取和RT-PCR对贝类中的甲肝病毒进行检测的方法。经比较,采用小体系肠道样品检测比采用全贝检测的富集效果更佳,并比较了PEG8000和PEG6000对病毒富集的效果,回收率分别为13.5%和7.6%,此方法可有效地降低PCR抑制剂的影响,最低检测限可达10个TCID50/1.5 g。

Study on the Detection of Hepatitis a Virus in Shellfish by RT-PCR

Hepatitis A is one of the major infectious diseases epidemiologically associated with worldwide shellfish consumption.The efficacy of method using polymerase chain reaction(PCR) to detect hepatitis A virus(HAV) in contaminated shellfish can be hindered by low recoveries during the concentration process and by natural PCR inhibitors in shellfish.The method for the detection of Hepatitis A virus from shellfish by twice polyethylene glycol(PEG)-precipitation,extraction of virus RNA and RT-PCR was developed in the report.The comparison of two different methods for sampling showed that the small system with the stomachs and digestive diverticula was more effective than the large system with whole shellfish tissue.The effect of virus concentration from shellfish by PEG8000 was compared with that by PEG6000,and the recovery was 13.5% and 7.6% respectively.The method developed can reduce the inhibition of PCR inhibitors and the detection limit was 10TCID_(50)/1.5g.