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骨组织中聚合酶对聚合酶链反应抑制剂的抗性*

Polymerase resistance to polymerase chain reaction inhibitors in bone*.

作者信息

Eilert Kenneth D, Foran David R

机构信息

Forensic Science Program, School of Criminal Justice, Michigan State University, East Lansing, MI 48824, USA.

出版信息

J Forensic Sci. 2009 Sep;54(5):1001-7. doi: 10.1111/j.1556-4029.2009.01116.x. Epub 2009 Aug 3.

Abstract

Amplification of DNA from aged or degraded skeletal remains can be a challenging task, in part due to naturally occurring inhibitors of the polymerase chain reaction. PCR inhibitors may act by inactivating a polymerase itself, or compete with or bind other reaction components, although various polymerases may be differentially susceptible to such insult. In this study, ten thermostable polymerases from six bacterial species were examined for their ability to amplify DNA in the presence of bone-derived or individual PCR inhibitors. Two polymerases, one from Thermus aquaticus and one from Thermus thermophilus, showed lower susceptibility to inhibition from bone, while polymerases from Thermus flavus were highly susceptible. Addition of bovine serum albumin improved the activity of most of the enzymes. Taken together, the results indicate that thermostable DNA polymerases have different susceptibility to bone-derived PCR inhibitors, and that those most often used in forensic laboratories may not be optimal when working with DNA from skeletal remains.

摘要

从老化或降解的骨骼遗骸中扩增DNA可能是一项具有挑战性的任务,部分原因是聚合酶链反应存在天然抑制剂。PCR抑制剂可能通过使聚合酶本身失活起作用,或者与其他反应成分竞争或结合,尽管各种聚合酶对此类损害的敏感性可能不同。在本研究中,检测了来自六种细菌的十种热稳定聚合酶在存在骨源或单独PCR抑制剂的情况下扩增DNA的能力。两种聚合酶,一种来自嗜热水生栖热菌,一种来自嗜热栖热菌,对来自骨骼的抑制作用敏感性较低,而来自黄栖热菌的聚合酶高度敏感。添加牛血清白蛋白可提高大多数酶的活性。综合来看,结果表明热稳定DNA聚合酶对骨源PCR抑制剂的敏感性不同,并且在处理骨骼遗骸的DNA时,法医实验室最常用的那些聚合酶可能并非最佳选择。

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