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目的了解输入性恶性疟原虫氯喹抗性转运蛋白基因(Pfcrt基因)K76T点突变发生情况。方法采集2008-2012年从非洲、东南亚等疟疾流行区回国人员恶性疟现症患者血样,根据恶性疟原虫Pfcrt基因序列设计巢式PCR引物,以血样中的恶性疟原虫DNA为模板,进行巢式PCR RFLP分析。结果92份输入性恶性疟现症患者血样中,突变型Pfcrt基因50份,占54.3%;野生型Pfcrt基因42份,占45.7%。采自非洲国家(尼日利亚、赤道几内亚、利比里亚等9国)回国人员的70份血样中,野生型Pfcrt基因37份,突变型Pfcrt基因33份,分别占52.9%和47.1%;采自东南亚国家(缅甸和印度尼西亚)回国人员的22份血样中,野生型Pfcrt基因5份,突变型Pfcrt基因17份,分别占22.7%和77.3%。不同地区回国人员Pfcrt基因突变发生率差异有统计学意义(χ2=6.12,P<0.05)。结论来自不同流行区的恶性疟原虫分离株Pfcrt基因突变发生率也不同,Pfcrt基因K76T位点突变检测在输入性恶性疟原虫氯喹抗性监测中具有应用价值。
Objective To investigate the incidence of K76T point mutation in the input gene of Plasmodium falciparum chloroquine resistance transporter (Pfcrt gene). Methods Blood samples of falciparum malaria patients returning from malaria-endemic areas in Africa and Southeast Asia from 2008 to 2012 were collected. According to the Pfcrt gene sequence of Plasmodium falciparum, nested PCR primers were designed. Plasmodium falciparum DNA in blood samples was used as a template for nested PCR RFLP analysis. Results Of the 92 cases of imported P. falciparum, 50 were mutated Pfcrt genes (54.3%) and 42 were wild-type Pfcrt genes (45.7%). Of the 70 blood samples taken from returnees in African countries (Nigeria, Equatorial Guinea, Liberia and other 9 countries), 37 were wild-type Pfcrt genes and 33 were Pfcrt mutations, accounting for 52.9% and 47.1% of the total. Blood samples collected from Southeast Asian countries Burma and Indonesia), 5 were wild-type Pfcrt genes and 17 were mutated Pfcrt genes, accounting for 22.7% and 77.3% respectively. The incidence of Pfcrt gene mutation in returnees in different regions was significantly different (χ2 = 6.12, P <0.05). Conclusions The incidence of Pfcrt gene mutations in P. falciparum isolates from different endemic areas is also different. The detection of Pfcrt gene K76T mutation has the value of monitoring chloroquine resistance in Plasmodium falciparum.