论文部分内容阅读
半知菌亚门中的念株菌(Candida)约有20个种是人体呼吸道,胃肠道,皮肤及生殖器官的常见腐生菌,它们还是AIDS患者念株菌性鹅口疮、食道炎及侵袭性感染常见的原因.通常的血清学鉴定方法是以不同菌体表面多糖抗原物质的差异为依据,但由于念珠菌有些菌体间抗原物质很相似,相互间有交叉反应,因此很难反映出实际的感染情况;而传统的表型分型法除了受环境因素和真菌本身变异的影响外,还受到实验本身重复性差,操作过程繁琐以及实验结果的判断有时并不十分明确等诸多因素的干扰.因此,建立一种稳定、快速、准确的致病性念珠菌基因分型鉴定方法在临床上对指导诊断和治疗具有重要的意义.由Zabeau等人1992年创立的AFLP(Amplified fragment length polymorphism)技术是目前国际上构建DNA指纹图谱的最新方法之一,该方法结合了RFLP(Restriction frag-ment length polymorphism)技术和RAPD(Random amplified polymorphic DNA)技术的特点,使用人工接头(Adapter)与限制性内切酶酶切的基因组DNA片段连接并以此为DNA模板,合成系列3’-末端随机变化数个碱基且与人工接头序列相互补的PCR引物进行PCR特异条件扩增,经电泳检测后可获得DNA指纹图谱.本研究针对临床上常见的8种致病性念珠菌,以Pst I酶切基因组DNA构建了致病性念株
There are about 20 Candida species in the Candida, which are common saprophytic bacteria in the human respiratory tract, gastrointestinal tract, skin and reproductive organs. They are also endemic to thrush, esophagitis and esophagitis in AIDS patients common cause infections, usually of serological identification method is based on differences in different cell surface antigen polysaccharide based material, but due to Candida species among some bacterial antigens are similar, there are cross-reactive with each other, it is difficult to reflect The actual situation of infection; and the traditional phenotyping method in addition to environmental factors and fungal mutation itself, but also by the poor reproducibility of the experiment itself, the operation is cumbersome and the results of the experiment is sometimes not very clear judgment and many other factors Therefore, the establishment of a stable, fast and accurate genotyping pathogenic Candida identification methods has important significance in guiding diagnosis and treatment in clinical practice. Zabeau and others founded by the 1992 AFLP (Amplified fragment length polymorphism) Technology is currently one of the latest international methods to construct DNA fingerprinting, which combines the RFLP (Restriction frag-ment length polymorphism) technology and RAP D (Random amplified polymorphic DNA) is characterized by the use of artificial adapter (Adapter) and restriction endonuclease digested genomic DNA fragments and as a DNA template, a series of synthetic 3’-end random number of bases and with artificial linker sequence complementary to PCR primers specific for PCR amplification conditions, after electrophoresis DNA fingerprints obtained in this study for 8 kinds of common pathogenic Candida clinically, Pst I digested genomic DNA was constructed Pathogenic concept strain