恶性疟原虫疫苗的研究──Ⅰ．环子孢子蛋白基因中央保守区的体外扩增

STUDY ON PLASMODIUM FALCIPARUM VACCINE──I.IN VITRO AMPLIFICATION OF THE CENTRAL CONSERVATIVE REGION OF CIRCUMSPOROZOITE PROTEIN GENE

恶性疟原虫环子孢子蛋白基因中央保守区编码产物具有３７个ＮＡＮＰ串联重复序列，并认为（ＮＡＮＰ）ｎ具有保护性免疫作用。本文通过酚／氯仿抽提乙醇沉淀方法提取恶性疟原虫全基因组ＤＮＡ，首次采用ＰＣＲ方法获得环子孢子蛋白ＣＳＰ基因中央保守区片段。我们用地高辛标记的ＰＣＲ产物作ＤＮＡ探针，杂交试验表明具有高度的敏感性和特异性。同时，本文还探讨了ＰＣＲ技术在病原学检测、疫苗研制、流行病学调查、疗效考核等方面的潜在应用前景。

It is very interesting field for parasitologists to study circumsporozoite protein(CSP) of Plasmodium falciparum. The central conservative region of CSP gene encodes 37 tandem repetitive copies of a Asn-Ala-Asn-Pro(NANP) tetrapeptide, and it is recognized that(NANP)-[n] can bring protective immunity to hosts. In this report, genomic DNA of the parasite protozoan is extracted with phenol /chloroform and precipitated with ethanol, and we successfully gain the fragments of the central conservative region of CSP gene using PCR technique. We have labelled PCR production with digoxigenin as DNA probe, the gene hybridization assay shows that it is both sensitive and specific. In this report, the latent application of PCR are discussed in detection of Plasmodium falciparum, research of vaccine, epidemical investigation, assessment of the effect of drug treatments, etc. The project supported by National Natural Science Foundation of China.