[Detection of Plasmodium falciparum infection in humans by (Dig)-DNA probe]

Abstract

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' blood samples, 33 hybridized positively with this probe. The positivity rate was 86.8%. Twenty-one normal blood 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.