Joly S, Pujol C, Schröppel K, Soll D R
Department of Biological Sciences, University of Iowa, Iowa City 52242, USA.
J Clin Microbiol. 1996 Dec;34(12):3063-71. doi: 10.1128/jcm.34.12.3063-3071.1996.
Candida tropicalis has emerged as the second most frequent colonizing Candida species, and it has been documented in nosocomial infections. To develop an effective fingerprinting system for this species, which is amenable to computer-assisted analyses and which provides information on the genetic relatedness of independent isolates, two DNA fragments, Ct3 (18,000 bp) and Ct14 (20,000 bp), were cloned from a genomic library of Sau3AI partial digestion products. Both probes generate complex Southern blot patterns containing 8 to 20 bands, when hybridized to EcoRI- or EcoRI-HaeIII-digested DNA of independent C. tropicalis isolates. The two probes show no cross-hybridization and are both species specific for C. tropicalis. A comparison of the capacity of the two probes to identify the same strain in different isolates, and differentiate unrelated strains, using computer-assisted computation of similarity coefficients and the genesis of dendrograms, suggests that while Ct14 is more effective in grouping independent isolates, Ct3 is more effective in discriminating small differences in the patterns of highly related isolates and is therefore a more effective probe for determining microevolution within a clonal population and substrain shuffling in recurrent infections. Because of their alternative attributes, it is suggested that both probes be used in fingerprinting studies of C. tropicalis.
热带念珠菌已成为第二常见的定植念珠菌物种,并且已在医院感染中被记录。为了开发一种针对该物种的有效指纹识别系统,该系统适用于计算机辅助分析并能提供独立分离株的遗传相关性信息,从Sau3AI部分消化产物的基因组文库中克隆了两个DNA片段,Ct3(18,000 bp)和Ct14(20,000 bp)。当与独立的热带念珠菌分离株的EcoRI或EcoRI - HaeIII消化的DNA杂交时,这两种探针均产生包含8至20条带的复杂Southern印迹图谱。这两种探针没有交叉杂交,并且都是热带念珠菌的种特异性探针。使用计算机辅助计算相似系数和构建树状图,比较这两种探针在不同分离株中鉴定同一菌株以及区分不相关菌株的能力,结果表明,虽然Ct14在对独立分离株进行分组方面更有效,但Ct3在区分高度相关分离株模式中的微小差异方面更有效,因此是确定克隆群体内微进化和复发性感染中菌株改组的更有效探针。由于它们的不同特性,建议在热带念珠菌的指纹识别研究中同时使用这两种探针。