一种用于检测常见硫嘌呤甲基转移酶（TPMT）突变的多重等位基因特异性聚合酶链反应检测法。

A multiplexed allele-specific polymerase chain reaction assay for the detection of common thiopurine S-methyltransferase (TPMT) mutations.

作者信息

Roberts Rebecca L, Barclay Murray L, Gearry Richard B, Kennedy Martin A

机构信息

Department of Pathology, Christchurch School of Medicine and Health Sciences, University of Otago, PO Box 4345, Christchurch, New Zealand.

出版信息

Clin Chim Acta. 2004 Mar;341(1-2):49-53. doi: 10.1016/j.cccn.2003.10.029.

DOI:10.1016/j.cccn.2003.10.029

PMID:14967158

Abstract

Thiopurine S-methyltransferase (TPMT) catalyses the S-methylation of thiopurine drugs that are commonly used to treat a wide range of conditions. It is now well established that interpatient variation in sensitivity to these drugs is due to point mutations in the TPMT gene. The mutant alleles TPMT2 (238G>C), TPMT3A (460G>A, 719A>G), TPMT3B (460G>A), and TPMT3C (719A>G) account for 80-95% of TPMT deficiency observed in Caucasian populations. In this paper, we describe a novel, multiplex, allele-specific polymerase chain reaction (PCR) method that detects the 238G>C, 460G>A, and 719A>G mutations, allowing for the simultaneous identification of TPMT2 and TPMT3 alleles. The assay is internally controlled, robust, and does not require the use of restriction endonucleases. Therefore, the assay is not prone to erroneous readings due to incomplete restriction digestion, as documented for existing PCR restriction fragment length polymorphism (RFLP) assays of TPMT.

摘要

硫嘌呤S-甲基转移酶（TPMT）催化常用于治疗多种病症的硫嘌呤类药物的S-甲基化反应。现已明确，患者对这些药物敏感性的个体差异是由TPMT基因中的点突变所致。突变等位基因TPMT2（238G>C）、TPMT3A（460G>A，719A>G）、TPMT3B（460G>A）和TPMT3C（719A>G）占白种人群中观察到的TPMT缺乏症的80 - 95%。在本文中，我们描述了一种新型的多重等位基因特异性聚合酶链反应（PCR）方法，该方法可检测238G>C、460G>A和719A>G突变，从而能够同时鉴定TPMT2和TPMT3等位基因。该检测方法有内部对照，稳健可靠，且无需使用限制性内切酶。因此，与现有的TPMT的PCR限制性片段长度多态性（RFLP）检测方法不同，该检测方法不会因限制性消化不完全而出现错误读数。