摘要
应用ＰＣＲ－ＳＳＣＰ银染技术，对皮肤利什曼病（ＣＬ）病人（Ｐ１７）的病原体ＳＳＵｒＲＮＡ基因及ｋＤＮＡ进行分析。将新疆克拉玛依地区皮肤利什曼病患者病变组织及有关利什曼原虫种株，Ｌ．ｉｎｆａｎｔｕｍ，Ｌ．ｔｒｏｐｉｃａ，Ｌ．ｄｏｎｏｖａｎｉＸｉｎｊｉａｎｇ“７７１”株，分别抽提各标本的基因组ＤＮＡ和ｋＤＮＡ后，再用相应的引物，Ｒ２２２和Ｒ３３３ＰＣＲ扩增ＳＳＵｒＲＮＡ基因（３９７ｂｐ片段）和１３Ａ、１３Ｂ扩增ｋＤＮＡ微环（１２０ｂｐ片段）。ＰＣＲ扩增的ＳＳＵｒＲＮＡ基因（３９７ｂｐ片段）经非变性聚丙烯酰胺凝胶电泳，银染显色后，ＣＬＰ１７病原体的ｓｓＤＮＡ的迁移率与Ｌ．ｔｒｏｐｉｃａ者接近，而与Ｌ．ｉｎｆａｎｔｕｍ者亦相差较小。ＰＣＲ扩增的ＣＬＰ１７病原体和Ｌ．ｉｎｆａｎｔｕｍ的ｋＤＮＡ微环（１２０ｂｐ）片段，经ＳＳＣＰ分析，ＣＬＰ１７病原体的Ｍｃ１和Ｍｃ２分别为０．８２５９、０．７２２２，Ｌ．ｉｎｆａｎｔｕｍ的ＭＦ１和ＭＦ２分别为０．９０７４、０．７９６２。ＭＣ与ＭＦ相比，二者相差较小；ＣＬＰ１７病原体和Ｌ．ｔｒｏｐｉｃａ的ｋＤＮＡ微环（１２０ｂｐ片段），经ＳＳＣＰ分析，ＣＬＰ１７病原体的ＭＣ１和ＭＣ２分别为?
PCR SSCP silver staining technique was applied to analyze the SSU rRNA gene and kDNA of cutaneous leishmaniasis pathogen (CLP) from Karamy,Xinjiang and related leishmania species:L-infantum L.tropica and L-donovani Xinjiang “771” strain.The 397bp fragment of SSU rDNA and 120bp fragment of kDNA were amplified with primers R222,R333 and 13A,13B respectively.The amplified fragments of CLP and related Leishmania species were run in 8% non denaturing mini polyacrylamide gel and then silver stained.The mobility of ssDNA of SSU rDNA from CLP was closer to that from Ltropica than that from Linfantum.The SSCP analysis of kDNA amplified fragments showed that the mobility of the first and the second ssDNA of kDNA from CLP17 was MC1=0.8259 and MC2=0.7222 respectively,the mobility of those from Linfantum was Mf1=09074 and Mf2=0.7962 respectively,the differences between MC1 and Mf1,MC2 and Mf2were shown .The SSCP analysis of DNA amplified fragments showed that the mobility of those from CLP was MC1=0.8448 and Mc2=0.7069,the mobility of those from L-tropica was Mt1=0.8448 and Mt2=0.7241 respectively,their mobility were quit similar.All these indicatied that there is a certain degree of similarity among CLP17,L-tropica,L-infantum;but CLP17 is different from L-tropica and L-infantum.
出处
《寄生虫与医学昆虫学报》
CAS
1998年第3期142-147,共6页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家教委博士学科点专项基金
国家自然科学基金
