合成酶联DNA探针对恶性疟病人的检测

A STUDY OF ENZYME-CONJUGATED SYNTHETIC DNA PROBE FOR THE DIAGNOSIS OF FALCIPARUM MALARIA

应用一种合成的P.f特异的21个硷基的酶联DNA探针(PFRl—AP),对培养的海南株P.f DNA及恶性疟病人血样进行检测。DNA杂交结果表明,该探针检测纯化的P.f DNA最低限度达8pg;62例经血片镜检确诊的P.f病人血样,39例杂交反应阳性,阳性率为75％。如将阴性病例的检测血样增加至100μl,则累计阳性率可达88.46％。3例P.f+P.v混合感染者杂交反应阳性。5例P.v患者血样为阴性反应,32例正常人亦未出现杂交反应。本研究结果提示,应用该探针检测有较高的敏感性,可与^(32)P标记的P.f基因群DNA或重组DNA的现场检测结果相近似,而且对P.v及正常人DNA无交叉反应,特异性也好。

A 21-base, Plasmodium falciparum specific, enzyme-conjugated synthetic DNA probe (PFRl-AP) was used for the diagnosis of falciparum malaria. The blood samples (53 P. falciparum, 5 P. vivax, and 3 P. f and P.v mixed infection cases ) collected from Hainan province were tested. The samples of 32 college students were used for normal control. The probe proved to be specific and sensitive. 10pg of purified P.f DNA could be always detected, and there was no cross reaction with the purified DNA of human leukocytes. When testing Hainan blood specimens, PFRl-AP specifically detected P.f infections. In dot blot, when Nytran membrane with 50 microliters of treated blood samples being used, 39 out of 52 P.f specimens hybridized with this probe positively. When the volume of blotted sample was increased to one hundred microliters, the accumulative total positive rate rose up to 88.46%. The samples of P.v and normal control showed negative reaction with this probe.