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使用快速ABO基因分型技术鉴定法医类型样本上的ABO等位基因。

Identification of ABO alleles on forensic-type specimens using rapid-ABO genotyping.

作者信息

Crouse C, Vincek V

机构信息

Palm Beach County Sheriff's Office Crime Laboratory, FL, USA.

出版信息

Biotechniques. 1995 Mar;18(3):478-83.

PMID:7779399
Abstract

Historically, forensic and clinical laboratories utilize serological techniques to identify ABO blood types. These techniques rely on the detection of ABO-associated proteins and are sensitive with very accurate results. This laboratory has simplified the identification of ABO types by taking advantage of previously reported ABO DNA sequence differences. The Rapid-ABO technique involves a two-step process: (i) amplification of DNA samples using primer sets specific for the ABO alleles and (ii) electrophoresis and visualization of amplified ABO fragments on a 3% MetaPhor agarose gel. The major advantage of the Rapid-ABO technique is the identification of ABO genotypes compared to serological tests for ABO phenotypes. This two-step process identifies six possible ABO genotypes including AB, AA, BB, AO, BO and OO. The Rapid-ABO protocol works well with DNA extracted organically or using Chelex 100. Results can be obtained in less than a day utilizing 2 ng of DNA in the amplification reaction. Analysis of 23 animal species shows the Rapid-ABO primers amplify ABO alleles from only human, chimpanzee and gorilla DNA.

摘要

从历史上看,法医和临床实验室利用血清学技术来鉴定ABO血型。这些技术依赖于对ABO相关蛋白质的检测,灵敏度高且结果非常准确。本实验室利用先前报道的ABO DNA序列差异简化了ABO血型鉴定。快速ABO技术包括两个步骤:(i)使用针对ABO等位基因的引物对DNA样本进行扩增,以及(ii)在3%的MetaPhor琼脂糖凝胶上对扩增的ABO片段进行电泳和可视化。与ABO血型血清学检测相比,快速ABO技术的主要优势在于能够鉴定ABO基因型。这个两步过程可鉴定出六种可能的ABO基因型,包括AB、AA、BB、AO、BO和OO。快速ABO方案对有机提取的DNA或使用Chelex 100提取的DNA都适用。在扩增反应中使用2 ng DNA,不到一天就能获得结果。对23种动物物种的分析表明,快速ABO引物仅能从人类、黑猩猩和大猩猩的DNA中扩增出ABO等位基因。

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