López-Sabater E I, Deng M Y, Cliver D O
Department of Food Microbiology and Toxicology, WHO Collaborating Center on Food Virology, University of Wisconsin-Madison, USA.
Lett Appl Microbiol. 1997 Feb;24(2):101-4. doi: 10.1046/j.1472-765x.1997.00357.x.
In order to detect the low numbers of hepatitis A viral (HAV) particles which may potentially be present in food and cause a serious illness, an original procedure which combines immunomagnetic separation and PCR is described. The use of streptavidin magnetic beads coated with biotinylated human anti-HAV IgG allows virus capture and the removal of the RT-PCR inhibitory compounds which usually are present in shellfish extracts. Following immunomagnetic capture, the separated HAV were lysed, the beads discarded, and the supernatant containing the viral RNA subjected to the RT-PCR protocol. Levels of HAV ranging from 10 to 10(5) pfu were successfully detected in artificially contaminated samples of shucked American oyster (Crassostrea virginica).
为了检测食品中可能存在的少量甲型肝炎病毒（HAV）颗粒，这些颗粒可能会引发严重疾病，本文描述了一种将免疫磁分离和聚合酶链反应（PCR）相结合的原创方法。使用包被有生物素化人抗HAV IgG的链霉亲和素磁珠可实现病毒捕获，并去除通常存在于贝类提取物中的逆转录聚合酶链反应（RT-PCR）抑制性化合物。免疫磁捕获后，裂解分离出的HAV，丢弃磁珠，将含有病毒RNA的上清液进行RT-PCR检测。在人工污染的去壳美国牡蛎（弗吉尼亚巨蛎）样本中成功检测到了10至10⁵ 空斑形成单位（pfu）的HAV水平。
