Ruan Li, Zhao Huian, Li Qingge
Molecular Diagnostics Laboratory, The Key Laboratory of The Ministry of Education for Cell Biology and Tumor Cell Engineering, Department of Biomedicine, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China.
J Forensic Sci. 2010 Jan;55(1):19-24. doi: 10.1111/j.1556-4029.2009.01228.x. Epub 2009 Dec 2.
Rapid and informative ABO genotyping has become increasingly popular in forensic use. We developed a multiplex real-time polymerase chain reaction (PCR) approach to genotype ABO major groups and subgroups. Seven differently fluorophor-labeled displacing probes for O(1)(261delG), A(261G), A(796C/803C), B(796A/803C), O(2) (802G>A), A(2) (1059delC), and A(2) (1009A>G) were combined in one or two PCRs to determine either ABO major groups or subgroups. The method correctly detected 13 reference DNA samples. A blind test of 237 samples resulted in complete agreement with their phenotypes, and 110 of these 237 samples as well as with PCR-SSP method. The whole analysis could be finished in less than 100 min at substantially low material cost and the template DNA ranging from 0.16 to 500 ng per reaction could be quantitatively detected. Despite the limited informativeness of ABO genotyping, the developed methods could find application in rapid and inexpensive screening of forensic settings.
快速且信息丰富的ABO基因分型在法医学应用中越来越受欢迎。我们开发了一种多重实时聚合酶链反应（PCR）方法来对ABO主要血型和亚型进行基因分型。七种不同荧光团标记的置换探针，分别针对O(1)(261delG)、A(261G)、A(796C/803C)、B(796A/803C)、O(2) (802G>A)、A(2) (1059delC)和A(2) (1009A>G)，在一个或两个PCR反应中组合使用，以确定ABO主要血型或亚型。该方法正确检测了13个参考DNA样本。对237个样本的盲测结果与它们的表型完全一致，并且这237个样本中的110个样本的结果也与PCR-SSP方法一致。整个分析可以在不到100分钟内完成，材料成本极低，每个反应中0.16至500 ng的模板DNA可以被定量检测。尽管ABO基因分型的信息量有限，但所开发的方法可用于法医学环境中的快速且廉价的筛查。
