间日疟原虫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 VIV AX

作　　者：万磊[1] 智刚[1] 陈培霞[1] 薛采芳[1] 姜绍谆[1] 

机构地区：[1]西安第四军医大学

出　　处：《中华微生物学和免疫学杂志》1994年第2期96-99,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金

摘　　要：根据已知序列，设计出一对含克隆位点的寡聚核苷酸引物，用于间日疟原虫ＳＳＵｒＤＮＡ特定片段扩增；采用双温度点ＰＣＲ法，从我国间日疟患者血样ＤＮＡ抽提物中，直接扩增出预期大小片段（约６４０ｂｐ），而培养红内期恶性疟原虫株、人源利什曼原虫株，人源弓形虫株及正常人ＤＮＡ样本均无此扩增带出现。扩增片段进一步经限制性酶切和Ｎｏｒｔｈｅｒｎ印迹杂交分析，证实确为间日疟原虫ＳＳＵｒＤＮＡ目的片段。According to known SSUrDNA sequence,oligonucleotide primer pairs specific to Plasmod ium and to P.ui-uax,which incorporated cloning sites HincⅡand PstⅠ,were defined respectively with computer programming.Thedesired DNA fragment size of P.vivax, about 640 base pairs in length ,was directly amplified by two temperaturepoint PCR from cenventionally extracted genomic DNA of blood sample of P.rirax malaria patient from Y unnanprovince of China. No such amplified band appeared from the DNA oxtracted from cultivated erythrocytic stage ofP. falciparum isolated (FCC/AH ,FCC1/HN and FCC/YN),L.donoxani,T.gondii and human.The SSUrDNAfragment amplified of P.vivax was identified with methods of digestion of restriction endonucleases and Northernblot hybridization.

关 键 词：间日 疟原虫 SSUrDNA 聚合酶链反应 

分 类 号：R382.31[医药卫生—医学寄生虫学]