Institute of Biotechnology, University of Caxias do Sul, R. Francisco G. Vargas 1130, Caxias do Sul, Rio Grande do Sul 95001-970, Brazil.
World J Microbiol Biotechnol. 2012 Oct;28(10):3007-13. doi: 10.1007/s11274-012-1111-5. Epub 2012 Jun 29.
Based on sequence alignment, oligonucleotide primers targeting the Aeromonas extracellular lipase gene were developed for PCR detection of member of the genus. A pair of primers designed for conserved regions of the gene amplified a 276 bp sequence in all Aeromonas species and tested strains, but did not have a positive result with other Gram-positive and Gram-negative bacteria, showing high specificity and sensitivity. Selective enrichment in alkaline peptone water, followed by centrifugation, and direct usage of cells suspension as template, detected initial populations of 10 c.f.u. ml⁻¹. Single-strand conformation polymorphism analysis of the PCR products allowed the characterization of Aeromonas strains with a high discriminatory power (Simpson's index = 0.988). The method presented here provides a useful tool for the rapid detection of Aeromonas and the characterization of Aeromonas isolates.
基于序列比对，针对气单胞菌细胞外脂肪酶基因设计了寡核苷酸引物，用于 PCR 检测该属成员。一对针对基因保守区域设计的引物在所有气单胞菌物种和测试菌株中扩增出 276 bp 的序列，但与其他革兰氏阳性和革兰氏阴性细菌没有阳性结果，表现出高度的特异性和敏感性。在碱性蛋白胨水中进行选择性富集，然后离心，直接使用细胞悬浮液作为模板，可检测到初始种群为 10 c.f.u. ml⁻¹。PCR 产物的单链构象多态性分析允许对气单胞菌菌株进行高区分能力的特征描述（辛普森指数=0.988）。本文提出的方法为快速检测气单胞菌和气单胞菌分离株的特征描述提供了有用的工具。
