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用于敏感嵌合体检测的单倍型计数:早期白血病复发检测的潜力

Haplotype Counting for Sensitive Chimerism Testing: Potential for Early Leukemia Relapse Detection.

作者信息

Debeljak Marija, Mocci Evelina, Morrison Max C, Pallavajjalla Aparna, Beierl Katie, Amiel Marie, Noë Michaël, Wood Laura D, Lin Ming-Tseh, Gocke Christopher D, Klein Alison P, Fuchs Ephraim J, Jones Richard J, Eshleman James R

机构信息

Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, Maryland.

Department of Oncology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, Maryland.

出版信息

J Mol Diagn. 2017 May;19(3):427-436. doi: 10.1016/j.jmoldx.2017.01.005.

Abstract

Fields of forensics, transplantation, and paternity rely on human identity testing. Currently, this is accomplished through amplification of microsatellites followed by capillary electrophoresis. An alternative and theoretically better approach uses multiple single-nucleotide polymorphisms located within a small region of DNA, a method we initially developed using HLA-A and called haplotype counting. Herein, we validated seven additional polymorphic loci, sequenced a total of 45 individuals from three of the 1000 Genomes populations (15 from each), and determined the number of haplotypes, heterozygosity, and polymorphic information content for each locus. In addition, we developed a multiplex PCR that amplifies five of these loci simultaneously. Using this strategy with a small cohort of leukemic patients who underwent allogeneic bone marrow transplantation, we first attempted to define a threshold (0.26% recipient) by examining seven patients who tested all donor and did not relapse. Although this initial threshold will need to be confirmed in a larger cohort, we detected increased recipient DNA above this threshold 90 to 145 days earlier than microsatellite positivity, and 127 to 142 days before clinical relapse in four of eight patients (50%). Haplotype counting using these novel loci may be useful for ultrasensitive detection in fields such as bone marrow transplantation, solid organ transplant rejection, patient identification, and forensics.

摘要

法医学、移植学和亲子鉴定领域都依赖于人类身份检测。目前,这是通过扩增微卫星然后进行毛细管电泳来完成的。一种替代的且理论上更好的方法是使用位于DNA小区域内的多个单核苷酸多态性,我们最初使用HLA - A开发了这种方法并称之为单倍型计数。在此,我们验证了另外七个多态性位点,对来自千人基因组计划中三个群体的共45个人(每个群体15人)进行了测序,并确定了每个位点的单倍型数量、杂合性和多态信息含量。此外,我们开发了一种多重PCR,可同时扩增其中五个位点。使用这种策略对一小群接受异基因骨髓移植的白血病患者进行检测,我们首先通过检查七名检测结果全为供体且未复发的患者来尝试确定一个阈值(0.26%受体)。尽管这个初始阈值需要在更大的队列中得到证实,但我们在八名患者中的四名(50%)中检测到,受体DNA超过该阈值的时间比微卫星阳性早90至145天,比临床复发早127至142天。使用这些新位点进行单倍型计数可能对骨髓移植、实体器官移植排斥反应、患者身份识别和法医学等领域的超灵敏检测有用。

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