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基于逆转录聚合酶链反应(RT-PCR)的贝类甲型肝炎病毒(HAV)检测方法比较。

A comparison of RT-PCR-based assays for the detection of HAV from shellfish.

作者信息

Di Pinto A, Conversano M C, Forte V T, La Salandra G, Montervino C, Tantillo G M

机构信息

Dipartimento di Sanità e Benessere degli Animali, Università degli Studi, Bari, Italy.

出版信息

New Microbiol. 2004 Apr;27(2):119-24.

Abstract

The hepatitis A virus (HAV) is the most common cause of viral infection linked to shellfish consumption. The lack of correlation between the fecal coliform indicators and the presence of enteric viruses in shellfish and their harvesting waters points to the need for molecular methods to detect viruses. We compared two RT-PCR based techniques currently available for the detection of the hepatitis A virus (HAV) in shellfish. Both approaches involve extraction of viral particles by glycine buffer and concentration of virus particles by one or two PEG precipitation steps. One procedure involves as RNA extraction method the use of oligo (dT) cellulose to select poly (A) RNA, and the other uses a system in which total RNA is bound on silica membrane. Comparison of the two RT-PCR based methods highlighted the efficiency of the first approach which is less time-consuming and technically demanding than the second.

摘要

甲型肝炎病毒(HAV)是与食用贝类相关的最常见病毒感染病因。贝类及其捕捞水域中的粪大肠菌群指标与肠道病毒的存在之间缺乏相关性,这表明需要采用分子方法来检测病毒。我们比较了目前可用于检测贝类中甲型肝炎病毒(HAV)的两种基于逆转录聚合酶链反应(RT-PCR)的技术。两种方法都涉及通过甘氨酸缓冲液提取病毒颗粒,并通过一到两个聚乙二醇(PEG)沉淀步骤浓缩病毒颗粒。一种方法使用寡聚(dT)纤维素选择聚(A)RNA作为RNA提取方法,另一种方法使用将总RNA结合在硅胶膜上的系统。对这两种基于RT-PCR的方法的比较突出了第一种方法的效率,该方法比第二种方法耗时更少且技术要求更低。

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