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首页> 外文期刊>Transactions of the Royal Society of Tropical Medicine and Hygiene >An efficient PCR--SSCP-based method for detection of a chloroquine resistance marker in the PfCRT gene of Plasmodium falciparum.
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An efficient PCR--SSCP-based method for detection of a chloroquine resistance marker in the PfCRT gene of Plasmodium falciparum.

机译：一种基于PCR-SSCP的高效方法，用于检测恶性疟原虫PfCRT基因中的氯喹抗性标记。

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摘要

The spread of chloroquine resistance throughout the world poses a major problem in combating malaria. In the present study, an efficient polymerase chain reaction-single strand conformational polymorphism (PCR--SSCP)-based assay detected the PfCRT K76T point mutation, which is a marker for chloroquine resistance. For the first time, we have used a PCR--SSCP-based technique to identify the mutation in a single-step labelling reaction during PCR and SSCP gel electrophoresis. This assay is 100% efficient, giving no false-positive or -negative results, and can be carried out within a short bench time. We have successfully analysed 120 natural isolates using the PCR-SSCP method for detection of the chloroquine resistance marker and found 91 of the 120 samples to show the PfCRT T76 mutation, and 71% (65 of the 91 samples) showed a positive correlation with chloroquine resistance from the clinical data of the patients. The PCR-SSCP technique can also be applied for the detection of new haplotypes of the PfCRTgene and surveillance of chloroquine-resistant malaria in malaria-endemic localities around the world.
机译：氯喹抗药性在世界范围内的传播构成了与疟疾作斗争的主要问题。在本研究中，一种有效的聚合酶链反应-单链构象多态性（PCR--SSCP）为基础的检测方法检测到PfCRT K76T点突变，该突变是氯喹抗性的标志物。我们首次使用基于PCR-SSCP的技术在PCR和SSCP凝胶电泳过程中的单步标记反应中鉴定突变。该测定是100％有效的，没有假阳性或阴性结果，并且可以在短的工作时间内进行。我们已经使用PCR-SSCP方法成功分析了120种天然分离株以检测氯喹抗性标记物，发现120个样品中有91个显示PfCRT T76突变，而71％（91个样品中的65个）与氯喹呈正相关从患者的临床数据抵抗。 PCR-SSCP技术还可用于检测PfCRT基因的新单倍型，并在世界范围内疟疾流行地区监测对氯喹耐药的疟疾。

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