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两种使用福尔马林固定石蜡包埋组织的 DNA 提取方法在聚合酶链反应中鉴定先天性巨细胞病毒感染的效用。

Utility of two DNA extraction methods using formalin-fixed paraffin-embedded tissues in identifying congenital cytomegalovirus infection by polymerase chain reaction.

机构信息

Departamento de Infectología e Inmunología, Instituto Nacional de Perinatología, México, México.

Subdirección de Servicios Auxiliares de Diagnóstico, Instituto Nacional de Perinatología, México, México.

出版信息

Diagn Microbiol Infect Dis. 2020 Aug;97(4):115075. doi: 10.1016/j.diagmicrobio.2020.115075. Epub 2020 May 11.

Abstract

Formalin-fixed paraffin-embedded (FFPE) tissues are a source of biological material for molecular studies; several methods to extract DNA from FFPE tissues have been reported. This process is challenging because of formaldehyde-induced cross-linking between proteins and DNA as well as molecule fragmentation when unbuffered formalin is used for fixation. Here, 2 methods for DNA extraction from FFPE tissues, based on a chelating resin and silica membrane columns, were modified and compared in their capacity to detect human cytomegalovirus (HCMV) in congenital infections. Both methods were tested on 121 samples of brain, lung, spleen, and liver derived from 36 deceased preterm newborns. Twelve patients were selected, and UL55 and UL75 HCMV genes were detected by polymerase chain reaction in 16/36 samples. These 2 methods represent a useful tool for DNA recovery from FFPE tissues and HCMV molecular identification with the advantage of low cost, minimal steps, minimal sample use, being solvent-free, and being easy to implement in the laboratory.

摘要

福尔马林固定石蜡包埋(FFPE)组织是分子研究用生物材料的来源;已经报道了几种从 FFPE 组织中提取 DNA 的方法。由于甲醛诱导的蛋白质和 DNA 之间的交联以及未缓冲的甲醛固定时分子片段化,该过程具有挑战性。在这里,基于螯合树脂和硅胶膜柱的 2 种从 FFPE 组织中提取 DNA 的方法经过修改,并在检测先天性感染中人巨细胞病毒(HCMV)的能力方面进行了比较。这两种方法均在 36 名已故早产儿的 121 个脑、肺、脾和肝样本上进行了测试。选择了 12 名患者,通过聚合酶链反应在 16/36 个样本中检测到 UL55 和 UL75 HCMV 基因。这两种方法代表了从 FFPE 组织中回收 DNA 和进行 HCMV 分子鉴定的有用工具,其优点是成本低、步骤少、样本用量少、无溶剂且易于在实验室中实施。

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