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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Eco RⅤ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。
Objective To understand the mutation of related sites of Plasmodium falciparum multidrug resistance 1 (Pfmdr1) in Wuhan. Methods From 2010 to 2015, blood samples of patients diagnosed as falciparum malaria from returning workers in malaria-endemic areas such as Africa and Myanmar were collected. The nested PCR primers were designed according to the Pfmdr1 86,1042 and 1246 gene loci of P. falciparum, and the Pfmdr1 gene of P. falciparum was amplified by nested PCR. The amplified products were digested with restriction endonucleases ApoⅠ, AseⅠand Eco RⅤ respectively. The mutation rates of the 86,1042 and 1246 sites. Results A total of 187 blood samples were collected from patients with P. falciparum. All of them successfully amplified Pfmdr1 gene. The mutation rates of 86, 1042 and 1246 were 19.3% (36/187), 4.3% (8/187) and 9.6% 18/187). Among them, 175 were imported malaria blood samples from Africa, and the mutation rates of 86, 1042 and 1246 were 20.6% (36/175), 2.9% (5/175) and 10.3% (18/175), respectively; Myanmar imported malaria blood sample 12 3 cases detected 1042 sites, 86 sites were not detected and 1246 site mutation. Conclusion Point mutations were detected in the Pfmdr1 gene of Pfmdr1 gene of P. falciparum from Africa at point 104, 1042 of Pfmdr1 gene from P. falciparum in Myanmar.