质粒型pBF_4DNA探针用于疟疾诊断的研究

DETECTION OF MALARIA PARASITE IN BLOOD SAMPLES USING PLASMID pBF4 DNA PROBE

质粒pBF_4DNA 用^(32)P 标记后,与现场采集的50份疟疾病人血样进行斑点杂交,9份镜检为恶性疟者杂交全为阳性,与镜检符合率为100%;41份镜检为间日疟的标本,有3份出现杂交弱阳性;10份正常人血两者检查均为阴性。DNA 探针检测恶性疟原虫的灵敏度为0.001%原虫血症。血样存放2年之久,仍可用于 DNA 杂交。

Clone pBF4 was selected from Plasmodium falciparum genomic DNA library and contained DNA se-quences which were highly repeated within the parasite genome.The recombinant plasmid pBF4 DNA was ex-tracted and labelled with ^(32)p and hybridized with malaria samples from endemic areas of Yunnan province.A-mong 9 eases of P.falciparum determined by microscopy all of them demonstrated positive by probe hy-bridization,of 41 cases of P.vivax determined by microscopy 3 demonstrated weak positive of P.falciparumby hybridization.Ten normal human blood samples were negative both in microscopy and hybridization,thehybridized sensitivity of the probe with cultured parasite was 0.001% parasiteamia.The stude also showedthat the samples stored at -70℃ for 2 years still could be used for DNA hybridization.