Novel approach to quantitative reverse transcription PCR assay of mRNA component in autopsy material using the TaqMan fluorogenic detection system: dynamics of pulmonary surfactant apoprotein A

被引：25

作者：

Ishida, K ^{[1
]}

Zhu, BL ^{[1
]}

Maeda, H ^{[1
]}

机构：

[1] Osaka City Univ, Sch Med, Dept Legal Med, Abeno Ku, Osaka 5458585, Japan

来源：

FORENSIC SCIENCE INTERNATIONAL | 2000年 / 113卷 / 1-3期

关键词：

forensic pathology; quantitative reverse transcription-PCR; TaqMan fluorogenic detection system; surfactant apoprotein A; real-time PCR;

D O I：

10.1016/S0379-0738(00)00252-8

中图分类号：

DF [法律]; D9 [法律]; R [医药、卫生];

学科分类号：

0301 ; 10 ;

摘要：

A novel approach to quantitative reverse transcription (RT)-PCR assay of mRNA component using fluorescent TaqMan methodology and a new instrument (ABI Prism 7700 sequence detection system) was developed for autopsy materials. Pulmonary surfactant. apoprotein A (SP-A) mRNA from a cadaveric lung was quantitated in real-time. The target SP-A gene and the endogenous reference of glyceraldehyde-3-phosphate (GAPDH) were amplified in the same tube, and an amount of the target was normalized to the reference. This assay had a high reproducibility and discrimination even in forensic autopsy materials up to 96 h postmortem. Elevated SP-A expressions were determined in some cases. This system without post-PCR sample handling would be a very useful tool in pathological diagnosis and DNA analysis. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.

引用

页码：127 / 131

页数：5

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