Ha Ji-Hyoung, Choi Changsun, Ha Sang-Do
School of Food Science and Technology, Chung-Ang University, 4726 Seodongdaero, Ansung, Gyunggi-do, 456-756, Republic of Korea.
Department of Food Science and Technology, Chung-Ang University, 72-1 Nae-ri, Daeduk-myun, Ansung, Gyunggi-do, 456-756, Republic of Korea.
Food Environ Virol. 2014 Dec;6(4):290-6. doi: 10.1007/s12560-014-9156-2. Epub 2014 Jun 22.
Outbreaks of viral diseases are frequently associated with the consumption of minimally processed shellfish. Among the viruses in these outbreaks, hepatitis A virus (HAV) and human norovirus (NoV) have been increasingly reported as the most common food-borne pathogens. These viruses must be concentrated in tested samples in order to be detected. In this study, a method for the detection of NoV and HAV in shellfish using an immuno-magnetic separation (IMS) procedure combined with reverse transcriptase (RT)-PCR was developed. The IMS/RT-PCR method was applied to investigate the recovery rates of HAV, NoV GI.1, and GII.4 from oyster and mussel. Based on IMS/RT-PCR results, recovery rates for HAV from oyster and mussel test samples were 2.4 and 1.1%, respectively. The NoV GI.1 recovery rates from oyster and mussel samples were 4.9-9.2% (mean 6.9%) and 4.3-8.6% (mean 6.2%), respectively, and the NoV GII.4 recovery rates were 8.8 and 8.5%, respectively. These results verified that HAV, NoV GI.1, and GII.4 can be detected in all the test samples using the IMS/RT-PCR method, although the three inoculated viruses were recovered with low efficiency. In conclusion, the IMS/RT-PCR method can be used to efficiently and rapidly detect viruses such as HAV and NoV in shellfish such as oyster and mussel.
病毒性疾病的暴发常常与食用轻度加工的贝类有关。在这些暴发中出现的病毒里，甲型肝炎病毒（HAV）和人诺如病毒（NoV）作为最常见的食源性病原体，其报告日益增多。为了检测这些病毒，必须在检测样本中对其进行浓缩。在本研究中，开发了一种使用免疫磁珠分离（IMS）程序结合逆转录酶（RT）-PCR来检测贝类中NoV和HAV的方法。将IMS/RT-PCR方法应用于研究牡蛎和贻贝中HAV、NoV GI.1和GII.4的回收率。根据IMS/RT-PCR结果，牡蛎和贻贝检测样本中HAV的回收率分别为2.4%和1.1%。牡蛎和贻贝样本中NoV GI.1的回收率分别为4.9%-9.2%（平均6.9%）和4.3%-8.6%（平均6.2%），NoV GII.4的回收率分别为8.8%和8.5%。这些结果证实，尽管三种接种病毒的回收率较低，但使用IMS/RT-PCR方法可在所有检测样本中检测到HAV、NoV GI.1和GII.4。总之，IMS/RT-PCR方法可用于高效、快速地检测牡蛎和贻贝等贝类中的HAV和NoV等病毒。
