合肥市输入性恶性疟原虫Pfcrt基因K76T位点突变研究

Mutations of K76T positions of Pfcrt Gene in the Imported Plasmodium falciparum cases in Hefei

目的探讨合肥市输入性恶性疟原虫Pfcrt基因K76T位点的突变情况。方法采集2010年从非洲疟疾高流行区,务工回国输入性恶性疟病人血样11份,采用恶性疟原虫Pfcrt基因序列特异引物,以滤纸血片中的恶性疟原虫DNA为模板,进行巢氏PCR扩增基因,扩增产物经限制性内切酶酶切后鉴定。结果收集到11例滤纸血样,成功扩增6例,其中2例被酶切,为Pfcrt等位基因野生型;4例未被酶切,为Pfcrt等位基因突变型。结论合肥市输入性恶性疟中Pfcrt等位基因发生突变,应该加强输入性恶性疟的药物抗性监测。

Objective To investigate the mutations of the K76T position of Pfcrt gene of Plasmodium falciparum in the imported cases reported in Hefei.Methods Blood samples were collected from 11 patients infected with Plasmodium falciparum.A nested PCR（polymerase chain reaction） method was applied to amplify the Pfcrt gene and template DNA from Plasmodium falciparum in blood spot specimens.The amplified PCR products were evaluated after digestion by restriction enzyme.Results Six blood samples had been amplified successfully in the pool.Four of the six blood samples carrying the mutant allele at codon K76T were not digested,while the remaining 2 blood samples with wild allele K76T were digested.Conclusion The imported Plasmodium falciparum cases in Hefei have a high mutation rate of K76T position.Therefore,drug resistance in the imported Plasmodium falciparum treatment should deserve more attention in the future.