生物素化质粒PFrep20探针检测恶性疟原虫感染的实验研究  

Study on Detection of P.falciparum in Blood Samples with Biotiny lated Plasmid PFrep20 DNA Probe

作　　者：万磊[1] 陈培霞[1] Goman M 刘珺[1] 刘忠湘[1] 

机构地区：[1]第四军医大学寄生虫学教研室,西安市710032 [2]英国爱丁堡大学分子生物研究室

出　　处：《地方病通报》1993年第2期1-5,共5页Endemic Diseases Bulletin

摘　　要：Bio-11-dUTP经切口平移法标记于ρPFrep20探针,自身杂交敏感度为10ρg;检出血培养云南株Pf(恶性疟原虫)和病人血样中Pf密度低限分别为1/10~5和1/10~4(虫/红细胞);病人血样中最低Pf检出数为12000个;检测30份云南省恶性疟患者血样,28份阳性;检测20份云南省和海南省间日疟患者血样、29份正常人血样均为阴性;与三种动物疟原虫及人白细胞的DNA未发现杂交反应。表明ρPFrep20对云南株Pf具高度同源性和特异性,生物素化此探针检测Pf具较高敏感性。Plasmid PFrep20 DNA probe labeled with bio-11-dUTP by nick traslation was evaluated for detection of P.falciparum in blood samples. The hybridization sensitivity of biotinylated probe was at least 10 ρg, which was able to detect parasitemia level of 0.001% in 30μl of RBC from culture of P.falciparum (Yunnan isolate) and 0.01% in 30μl of RBC from p. alciparum malaria patients of Yunnan Province, smallest number of parasites detected were 12 000 in blood samples. With this probe 28 samples were obviously positive in detection of 30 blood samples from falciparum malaria patients of Yunnan Province, but detected 20 blood samples from vivax malaria patients of Yunnan and Hainan Provinces and 29 blood samples from donors of Xian all negative. In addition, this probe did not hybridize with the DNA of P.cynomolgi, P.knowlesi, P.yoelii and human leucocytes. It is demenstrated that plasmid PFrep20 probe is extremely homologous and specific to DNA of P.falciparum isolates of Yunnan Province.

关 键 词：生物素标记 探针 疟疾 诊断 

分 类 号：R531.304[医药卫生—内科学]