Identification of dual false indirect exclusions on the D5S818 and FGA loci
Identification of dual false indirect exclusions on the D5S818 and FGA loci
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DOI：
10.1016/j.legalmed.2010.08.006
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发表时间：
2011-01-01
期刊：
影响因子：
1.5
通讯作者：
Wang, Yue
中科院分区：
文献类型：
--
作者：
Jiang, Wenxiao;Kline, Margaret;Wang, Yue
在这里，我们提出了一个案例，其中的产妇测试的结果是模糊的，由于两个错误的间接排除发生在两个15遗传的孩子通过使用AmpFISTR Identifiler PCR扩增试剂盒（应用生物系统，福斯特市，CA）。Identifiler试剂盒未能扩增D5S818系统在孩子身上的等位基因11，也未能捕获母亲和孩子身上FGA系统上等位基因13的存在。这种情况通过使用Powerofiler 16 PCR扩增试剂盒（Promega，麦迪逊，WI）来补救，该试剂盒使用具有与Identifiler试剂盒不同的等位基因范围的不同引物。通过测序确认母性，发现Identifier试剂盒未能在D5S818系统上扩增等位基因11是由于重复序列下游55个碱基对（bp）处鸟嘌呤（G）变为胸腺嘧啶（T）的突变导致引物结合位点不相容的结果。Identifiler试剂盒不能拾取FGA系统的等位基因13是由于等位基因的位置超出范围。如果没有充分调查，间接排除可能会产生误导，因为等位基因范围以及引物结合亲和力是两个必须解决的混杂因素，以确保测试结果的准确性。(C)2010爱思唯尔爱尔兰有限公司版权所有。
Here, we present a case in which the result of a maternity test was obscured due to two false indirect exclusions that occurred in two out of 15 genetic kid through the use of the AmpFISTR Identifiler PCR Amplification kit (Applied Biosystems, Foster City, CA). The Identifiler kit failed to amplify allele 11 of the D5S818 system on the child and failed to capture the existence of allele 13 on the FGA system on both mother and child. The situation was remedied through use of the PowerPlex 16 PCR Amplification Kit (Promega, Madison, WI) which used different primers with a different allele range than that of the Identifiler kit. Maternity was confirmed through sequencing and it was found that the failure of the Identifier kit to amplify allele 11 on the D5S818 system was the result of an incompatibility to the primer-binding site due to a mutation that changed a guanine (G) into a thymine (T) 55 base pairs (bp) downstream of the repeat. The inability of the Identifiler kit to pick up allele 13 of the FGA system was due to the out-of-range location of the allele. Indirect exclusions can be misleading if they are not fully investigated since allele range as well as primer-binding affinity are two confounders that must be addressed to ensure accuracy of the test results. (C) 2010 Elsevier Ireland Ltd. All rights reserved.