恶性疟原虫SSUrDNA特定片段的体外扩增及鉴定  

AMPLIFICATION IN VITRO AND IDENTIFICATION OF SMALL SUBUNIT RIBOSOMAL DNA FRAGMENT OF PLASMODIUM FALCIPARUM 

作　　者：万磊[1,2] 陈培霞[1,2] 薛采芳[1,2] 刘忠湘[1,2] 姜绍谆[1,2] 

机构地区：[1]第四军医大学寄生虫学教研室 [2]第四军医大学微生物学教研室

出　　处：《中国寄生虫学与寄生虫病杂志》1994年第3期169-171,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金

摘　　要：根据已知疟原虫、其它寄生人体原虫及人的小亚单位核糖体核糖核酸基因（ＳＳＵｒＤＮＡ）编码区序列，借助计算机核酸序列软件分析，设计出一对用于恶性疟原虫（Ｐ．ｆ．）ＳＳＵｒＤＮＡ特定片段扩增的引物。采用双温度点聚合酶链反应（ＰＣＲ）技术，从我国Ｐ．ｆ．ＦＣＣ／ＹＮ茅芽株红内期基因组ＤＮＡ中，扩增出约５７０个碱基对（ｂｐ）的ＤＮＡ片段，与预期长度相符；而间日疟原虫（Ｐ．ｖ．）、利什曼原虫（Ｌ．ｄ．）、弓形虫（Ｔ．ｇ．）及人白细胞ＤＮＡ样本均无此扩增带出现。扩增片段经限制性内切酶消化和Ｎｏｒｔｈｅｒｎ印迹杂交分析，证实为Ｐ．ｆ．ＳＳＵｒＤＮＡ目的片段。According to computer analysis of SSUrDNA sequences of Plasmodium ,other protozooa and human,two oligonucleotide primers were designed.A DNA fragment,about 570 base pairs,was successfully amplified by two temperature point polymerase chain reaction from the genomic DNA of cultivated erythrocytic stage of P.falciparum FCC/YN(Simao),but no fragment was obtained from that of P.vivax,L.donovani,T.gondii and humans.It has been confirmed that the amplified fragment was indeed expected SSUrDNA segment of P.falciparum by means of restriction endonuclease digestions and Northern blot hybridization.

关 键 词：疟原虫 聚合酶链反应 SSUrDNA 

分 类 号：R382.31[医药卫生—医学寄生虫学]