摘要
采用异羟基洋地黄毒苷配基(Digoxigenin,(Dig)-11-dUTP),以随机引物法标记含有恶性疟原虫(P.f.)DNA的重组质粒片段(酶切克隆pPF14),制成pPF14-F-Dig探针。用此探针,以斑点杂交试验检测提纯的P.f.DNA及海南地区疟疾病人。结果显示,pPF14-F-Dig探针至少可检出40pg的P.f.DNA和低至0.005％原虫血症。38例P.f.病人中,检出阳性33例。3例恶性疟原虫和间日疟原虫混合感染者中,2例阳性。4例间日疟病人中,阴性3例,可疑1例。21例正常人血皆为阴性。
In this study, we used Digoxigenin, (Dig)-11-dUTP, in the Genius?kit purchased from Boehringer Mannheim to label the Plasmodium falciparum (P.f.) DNA fragment by random primed labelling method. The P. falciparum DNA fragment was isolated from the recombinant clone pPF14 by enzyme digestion and agarose electrophoresis. This probe is designated pPF14-F-Dig.Purified P.f. DNA and blood samples from malaria patients in Hainan Province of China were detected by the probe in dot-blot hybridization. Thirty-eight patients with falciparum malaria, three with falciparum malaria and vivax malaria and four with vivax malaria were detected with the pPF14-F-Dig probe.The results showed that the pPF14-F-Dig probe could detect at least 40 pg of P.f. DNA and a parasitemia of 0.005 percent. Out of 38 detected P.f.patients' bloot samples,33 hybridized positively with this probe.The positivity rate was 86.8%. Twenty-one normal bloot samples all showed negative. In addition, 2 of 3 samples with P. f. and P. v. were positive. Three of 4 samples with P.v. were negative and one was doubtful. The preliminary result of this study indicates that the pPF14-F-Dig probe could detect the patients of falciparum malaria with good sensitivity and specificity, and that this probe can be used to detect the patients with P. falciparum in Hainan Province.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
1991年第4期287-289,共3页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金
江苏省科委资助
关键词
疟原虫
DNA探针
疟疾
恶性
诊断
Plasmodium jalcipamm, falciparum malaria, (Dig)-DNA probe, hybridization
