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根据间日疟原虫环子孢子蛋白(CSP)基因序列,设计并合成间日疟原虫特异性引物一对,采用聚合酶链反应技术,进行间日疟原虫感染的检测。结果:(1)从间日疟原虫患者的全血DNA中扩增出约772bp的基因片段,经限制性内切酶切,证实为间日疟原虫CSP基因片段;(2)与间日疟原虫亲缘关系相近的三株恶性疟原虫、弓形虫、利什曼原虫和正常人全血核酸抽提物经扩增后均未见特异性的扩增条带;(3)该检测体系可检测出间日疟原虫感染血样中2.8虫/μl血的水平;(4)将该检测体系用于深圳地区及湖北随州地区间日疟感染患者血样的检测,147份患者的血样中144份检出阳性,可见一条特异性扩增带,与镜检的符合率达98%,而20正常人血样均为阴性。上述结果表明该法灵敏、特异且稳定性好,适用于间日疟感染的检测。
According to the sequence of Plasmodium vivax circumsporozoite protein (CSP) gene, a pair of specific primers were designed and synthesized. Plasmodium vivax was detected by polymerase chain reaction (PCR). Results: (1) The gene fragment of about 772bp was amplified from whole blood DNA of Plasmodium vivax patients and confirmed to be the CSP gene fragment of Plasmodium vivax by restriction enzyme digestion; (2) The three strains of Plasmodium falciparum, Toxoplasma gondii, Leishmania and normal human whole blood nucleic acid extracts with similar genetic relationship were not found any specific amplified bands. (3) The detection system (4) The detection system was applied to the blood samples of infected patients with P. vivax in Shenzhen area and Suizhou area of Hubei Province. 144 blood samples from 147 patients A positive detection showed a specific amplification band, with the coincidence rate of microscopic examination up to 98%, while 20 normal blood samples were negative. The above results show that the method is sensitive, specific and stable, suitable for the detection of P. vivax infection.