通过巢式逆转录聚合酶链反应检测贝类中的甲型肝炎病毒。
Detection of hepatitis A virus in shellfish by nested reverse transcription-PCR.
作者信息
Croci L, De Medici D, Morace G, Fiore A, Scalfaro C, Beneduce F, Toti L
机构信息
Laboratorio Alimenti, Istituto Superiore di Sanità, Rome, Italy.
出版信息
Int J Food Microbiol. 1999 Apr 1;48(1):67-71. doi: 10.1016/s0168-1605(99)00028-8.
A method for the detection of HAV in shellfish, based on the use of guanidinium isothiocyanate-containing solution for RNA extraction and purification steps, followed by nested PCR, is hereby proposed. Tests were carried out on mollusc samples spiked with HAV strain FG. Results showed that in samples subjected only to one round of PCR it was possible to detect HAV at concentrations of 10(3)-10(4) TCID50/10 g of mollusc. The use of the nested PCR renders the system more sensitive and specific enabling the identification of HAV concentrations as low as 1 TCID50/10 g of mollusc. Furthermore thus method, in addition to allowing the avoidance of confirming tests, such as hybridization, proved to be inexpensive and simple to perform.
本文提出一种检测贝类中甲型肝炎病毒（HAV）的方法，该方法在RNA提取和纯化步骤中使用含异硫氰酸胍的溶液，随后进行巢式聚合酶链反应（PCR）。对添加了HAV FG株的软体动物样本进行了测试。结果表明，在仅经过一轮PCR的样本中，能够检测到浓度为10(3)-10(4) 半数组织培养感染剂量（TCID50）/10克软体动物的HAV。使用巢式PCR使该系统更灵敏、特异，能够鉴定低至1 TCID50/10克软体动物的HAV浓度。此外，该方法除了无需进行杂交等确证试验外，还证明成本低廉且操作简单。
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