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目的研究中缅边境地区恶性疟原虫4个与抗疟药抗性相关基因的多态性,并与该地区以往检测结果进行比较,了解和分析该地区恶性疟原虫药物抗性新趋势。方法应用巢氏PCR或普通PCR方法检测中缅边境地区2014年恶性疟现症病人血样中恶性疟原虫4个抗药相关基因点突变情况,并与2007-2009年检测结果进行比较分析。结果该地区的恶性疟原虫DHFR基因51,59,108和164位点氨基酸有不同程度突变,其中三突变NRNL型为28.1%,IRNI型为10.0%,四突变IRNL型为61.9%,未见野生型和双突变。对PfDHPS的436,437,540和581位点的突变情况进行分析,仅见三突变,突变频率分别为SGEG型14.4%,SGNG型35.6%,AGEA型50.0%。对PfMDR1的86和1246位点进行分析,仅有86位点突变,频率为32%。未见1246位点突变。对PfATPase6分子的多态性分析显示全部为野生型,未见突变型。结论中缅边境地区恶性疟原虫抗性相关基因多态性与以往年比较有变化,其中DHFR、MDR1基因突变率升高,可为该地区恶性疟的防治提供参考。
Objective To study the polymorphisms of 4 genes related to resistance to Plasmodium falciparum in border areas of China and Burma and to compare with the previous detection results in this area to understand and analyze the new trend of drug resistance in Plasmodium falciparum in this area. Methods The mutations of four drug-resistance-related genes of Plasmodium falciparum in the blood samples of falciparum malaria patients in the Sino-Burma border areas were detected by nested PCR or ordinary PCR and compared with the results of 2007-2009. Results The amino acid residues 51, 59, 108 and 164 of DHFR gene of P. falciparum in this region were mutated to varying degrees. Among them, NRNL type of triple mutation was 28.1%, IRNI type was 10.0% and quadruple mutant type of IRNL was 61.9% Double mutation. The mutations of 436, 437, 540 and 581 in PfDHPS were analyzed. Only three mutations were found. The frequency of mutation was 14.4% for SGEG, 35.6% for SGNG and 50.0% for AGEA. Analysis of 86 and 1246 sites of PfMDR1 revealed only 86 mutations at a frequency of 32%. No mutation at point 1246 was found. Polymorphism analysis of the PfATPase6 molecule revealed that all were wild-type, with no mutant. Conclusion The polymorphisms of Plasmodium falciparum resistance-related genes in China-Myanmar border areas have changed compared with the past years. The mutation rate of DHFR and MDR1 genes is increased, which may provide reference for the prevention and treatment of P. falciparum in the area.