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使用环介导等温扩增技术(LAMP)对自然环境和实验室中各种物体表面血渍中人类男性DNA的时间进程检测比较

Comparison of Time Course Detection of Human Male DNA from Blood Stains on Various Objects on Surface in a Natural Environment and in a Laboratory Using Loop-Mediated Isothermal Amplification (LAMP).

作者信息

Kumsiri Ratchanok, Kanchanaphum Panan

机构信息

Pathobiology Unit, Faculty of Science, Rangsit University, Pathumthani, Thailand.

Biochemistry Unit, Faculty of Science, Rangsit University, Pathumthani, Thailand.

出版信息

Scientifica (Cairo). 2021 Nov 25;2021:4811608. doi: 10.1155/2021/4811608. eCollection 2021.

Abstract

In forensic study, the biological evidence can easily degrade, especially DNA. Degraded and environmentally challenged samples can produce numerous problems in forensic DNA analysis including loss of band product. Loop-mediated isothermal amplification or LAMP is one of the DNA analysis techniques used in forensic study. This study explores the limitations of the efficiency of the LAMP technique on abandoned DNA. For the DNA template, 8 male and 2 female blood-stained samples were taken from the surfaces, namely, brick, cloth, and tile from inside, and buried outside the laboratory. The LAMP reaction was used to amplify the SRY gene for detecting male DNA. All the blood-stained samples were stored for 1, 7, 15, 30, and 45 day (s). The LAMP product from the blood-stained samples on all the surfaces that were kept in a laboratory was detected using the gel electrophoresis technique from day 1 until day 45. However, the LAMP product on day 30 and 45 was smear and dim. The LAMP product from the blood-stained samples buried outside the laboratory was observed using the gel electrophoresis technique within day 30 (smear and dim). To increase the efficiency of detection, the qLAMP technique detected product on all the male samples from all the surfaces buried outside the laboratory for 45 days. The results indicate that this LAMP condition was possible detecting male DNA and the environmental factors are the main influence on the sensitivity of the LAMP technique. In addition, the qLAMP technique can increase the capacity and sensitivity of the detection.

摘要

在法医研究中,生物证据很容易降解,尤其是DNA。降解的以及受到环境挑战的样本会在法医DNA分析中产生诸多问题,包括条带产物的丢失。环介导等温扩增技术(LAMP)是法医研究中使用的DNA分析技术之一。本研究探讨了LAMP技术在处理废弃DNA时效率的局限性。对于DNA模板,从砖、布和瓷砖表面采集了8份男性和2份女性血样,分别取自室内和实验室外掩埋处。使用LAMP反应扩增SRY基因以检测男性DNA。所有血样均保存1、7、15、30和45天。从第1天到第45天,使用凝胶电泳技术检测保存在实验室的所有表面上血样的LAMP产物。然而,在第30天和第45天的LAMP产物出现拖尾且暗淡。使用凝胶电泳技术在30天内观察了埋在实验室外的血样的LAMP产物(拖尾且暗淡)。为了提高检测效率,定量LAMP技术在埋于实验室外45天的所有表面上的所有男性样本中均检测到了产物。结果表明,这种LAMP条件能够检测男性DNA,环境因素是影响LAMP技术灵敏度的主要因素。此外,定量LAMP技术可以提高检测能力和灵敏度。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a789/8639262/7f9078e9064b/SCIENTIFICA2021-4811608.001.jpg

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