用[α-^(32)P]—dATP标记恶性疟原虫DNA探针诊断恶性疟疾的研究

STUDIES ON (α-^(32)P)—dATP LABELLED DNA PROBE OF PLASMODIUM FALCIPARUM IN DIAGNOSIS OF FALCIPARUM MALARIA

本文报告从培养的恶性疟原虫提取基因组DNA,以[α-^(32)p]—dATP标记,作为探针,按DNA斑点杂交法检查取自海南省恶性疟患者的血样23份,结果全部呈杂交阳性反应,镜检阳性样本DNA探针检出率为100%,与10例间日疟患者血样全部出现交叉杂交。该探针可检测出恶性疟原虫密度为0.0001%的原虫血症水平,与鼠疟原虫、正常人血和人白细胞无交叉杂交。

The research work on the total genomic DNA probe of Plasmodium falciparum. in detection of malaria parasite in human blood was conducted. The total genomic DNA was isolated from cultured Fcc—1 isolates of Plasmodium falciparum and succeeding purification was done. The purified DNA was then labelled with domesticmade (α-^(32)p)-dATP through nick-translation. And subsequent DNA-DNA spot hybrid — ization was undertaken. the result of hybridization was evaluated by autoradiography. Twenty-three blood samples of patients suffering from Plasmodium falciparum.,from an endemic area of malaria in Hainan Province and ereyopreserved at —45℃ for 6 months were under study. Positive records were revealed in all the 23 samples, the compliance with microscopic blood examination being 100%. Cross hybridization was demonstrated in test of Plasmodium vivax samples while no evidence of cross reaction was exhibited in applying normal human blood, human leucocytes or rodent parasites including Plasmodium yoelii yoelii and Plasmodium berghei. The DNA probe was sensitive enough to detect as minimal as 0. 0001% parasitemia level in 20μl of blood sample.