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目的 :建立 PCR检测恶性疟原虫的新方法。方法 :作者采用自行设计并合成的一对恶性疟原虫特异引物 ,经 PCR扩增环子孢子蛋白 ( CSP)基因 3 端保守区序列 2 4 5bp片段 ,观察了它的特异性、敏感性和稳定性。结果 :1从 4株培养的恶性疟原虫和 2例恶性疟患者血样中均扩增出约 2 4 5bp的 DNA目的片段 ,用业已鉴定的 CSP基因序列作模板再行扩增证实其为恶性疟原虫CSP基因片段 ;2对间日疟原虫、利什曼原虫、弓形虫和健康人血样进行 PCR反应 ,均未见扩增条带 ;3本检测系统可检出恶性疟原虫感染血样中 0 .18个原虫所含的 DNA模板 ;4采用不同方法制备模板及不同反应方式均能获得满意结果 ;结论 :PCR扩增恶性疟原虫 CSP基因 3 端片段用于恶性疟原虫检测具有灵敏、高度特异且稳定性好等优点。
Objective: To establish a new method for PCR detection of Plasmodium falciparum. Methods: The authors designed and synthesized a pair of Plasmodium falciparum specific primers to amplify the 24 4p fragment of the 3-terminal conserved region of the circumsporozoite protein (CSP) gene by PCR and observed its specificity, sensitivity and stability Sex. RESULTS: A DNA fragment of about 24 bp was amplified from the blood samples of 4 strains of P. falciparum and 2 cases of P. falciparum. The amplified sequence of CSP gene was used as a template to confirm that it was falciparum malaria CSP gene fragment of protozoan; 2 pairs of PCR reactions between blood samples of Plasmodium vivax, Leishmania, Toxoplasma gondii and healthy people, no amplification bands; 3 detection system can detect Plasmodium falciparum infection blood samples 0. 18 protozoa contained DNA template; 4 using different methods to prepare the template and different reaction methods can get satisfactory results; Conclusion: PCR amplification of Plasmodium falciparum CSP gene 3-terminal fragment of Plasmodium falciparum for detection of sensitive, highly specific and Good stability and so on.