中国寄生虫学与寄生虫病杂志

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武汉市输入性恶性疟原虫多药抗性基因 1 相关位点突变分析

贾西帅,周水茂*,徐明星,杨燕,吴凯   

  1. 武汉市疾病预防控制中心,武汉430015
  • 出版日期:2016-12-30 发布日期:2017-01-10

Mutations of Plasmodium falciparum Multidrug Resistance 1 Gene in Imported Plasmodium falciparum in Wuhan

JIA Xi-shuai, ZHOU Shui-mao*, XU Ming-xing, YANG Yan, WU Kai   

  1. Wuhan Center For Disease Control and Prevention, Wuhan 430015, China
  • Online:2016-12-30 Published:2017-01-10

摘要:

目的 了解武汉市输入性恶性疟原虫多药抗性基因1(Plasmodium falciparum multidrug resistance 1,Pfmdr1)相关位点突变情况。 方法 2010-2015年,采集武汉市从非洲和缅甸等疟疾流行区回国人员中确诊为恶性疟的患者血样。根据恶性疟原虫Pfmdr1 86、1042和1246基因位点设计巢式PCR引物,巢式PCR扩增恶性疟原虫Pfmdr1基因,分别用限制性内切酶ApoⅠ、AseⅠ和EcoRⅤ酶切扩增产物,分析其86、1042和1246位点的突变率。 结果 共检测恶性疟现症患者血样187份,全部成功扩增出Pfmdr1基因,86、1042和1246位点突变率分别为19.3%(36/187)、4.3%(8/187)和9.6%(18/187)。其中非洲输入性疟疾血样175份,86、1042和1246位点突变率分别为20.6%(36/175)、2.9%(5/175)和10.3%(18/175);缅甸输入性疟疾血样12份,其中3例检出1042位点,未检出86位点和1246位点突变。 结论 来自非洲的输入性恶性疟原虫Pfmdr1基因86、1042和1246位点均检出点突变,来自缅甸的输入性恶性疟原虫Pfmdr1基因只检出1042位点发生突变。

Abstract:

Objective To identify Plasmodium falciparum multidrug resistance 1(Pfmdr1) point mutations in imported Plasmodium falciparum in Wuhan. Methods Blood samples were collected from returnees infected with P. falciparum in endemic areas of Africa and Myanmar during 2010-2015 in Wuhan City. Nested PCR primers were specifically designed for Pfmdr1 gene loci 86, 1042 and 1246 of P. falciparum. The Pfmdr1 gene was then amplified by nested PCR, and the products were digested by restriction enzyme ApoⅠ, AseⅠ and EcoRⅤ, respectively. The mutation rate for loci 86, 1042 and 1246 was analyzed. Results A total of 187 patients with falciparum malaria  were involved in the study. Pfmdr1 was amplified from all the blood samples. Restriction enzyme digestion revealed mutation rate of 19.3%(36/187), 4.3% (8/187) and 9.6%(18/187) for loci 86, 1042 and 1246, respectively. In detail, the mutation rate for loci 86, 1042 and 1246 was 20.6%(36/175), 2.9%(5/175) and 10.3%(18/175) respectively in the 175 samples from Africa, and only 3 cases with locus 1042 mutation were found in the 12 samples from Myanmar. Conclusion The loci 86, 1042 and 1246 mutations of Pfmdr1 have all been found in the samples from Africa, with only one point mutation (locus 1042) found in samples from Myanmar.

Key words: Plasmodium falciparum, Plasmodium falciparum multidrug resistance 1 gene, Polymorphism, Gene mutation