Maiti Biswajit, Raghunath Pendru, Karunasagar Iddya, Karunasagar Indrani
Department of Fishery Microbiology, UNESCO-MIRCEN for Marine Biotechnology, Karnataka Veterinary, Animal and Fisheries Sciences University, College of Fisheries, Mangalore, 575002, India.
J Microbiol Methods. 2009 Sep;78(3):312-8. doi: 10.1016/j.mimet.2009.07.009. Epub 2009 Jul 19.
Two PCR based typing methods i.e. random amplified polymorphic DNA analysis (RAPD) and enterobacterial repetitive intergenic consensus sequence (ERIC)-PCR were evaluated for typing of 42 Aeromonas isolates from clinical and environmental sources and whole cell protein (WCP) profiles were analyzed. Both RAPD and ERIC-PCR showed a high level of genetic diversity. Numerical index of the discriminatory (D) values were 0.94 and 0.96 (>0.90) for RAPD and ERIC-PCR, respectively. No correlation in banding pattern and evidence of genetic similarity was found between Aeromonas isolates from environmental and clinical sources. Therefore these techniques are highly reproducible and sensitive methods for typing the Aeromonas isolate from different sources. WCP profile showed two major variable regions i.e. 20 kDa to 45 kDa region and 70 kDa to 85 kDa region. Though WCP profiling had less discriminatory power, use of this method in combination with other established typing methods such as RAPD and ERIC-PCR may be helpful for reliable typing of Aeromonas isolates or to identify new proteins with pathogenic potential.
对两种基于聚合酶链反应(PCR)的分型方法,即随机扩增多态性DNA分析(RAPD)和肠杆菌基因间重复一致序列(ERIC)-PCR进行了评估,用于对42株来自临床和环境来源的气单胞菌进行分型,并分析了全细胞蛋白(WCP)图谱。RAPD和ERIC-PCR均显示出高度的遗传多样性。RAPD和ERIC-PCR的鉴别力(D)值的数值指标分别为0.94和0.96(>0.90)。在来自环境和临床来源的气单胞菌分离株之间,未发现条带模式的相关性和遗传相似性证据。因此,这些技术是用于对不同来源的气单胞菌分离株进行分型的高度可重复且灵敏的方法。WCP图谱显示出两个主要可变区,即20 kDa至45 kDa区域和70 kDa至85 kDa区域。尽管WCP图谱分析的鉴别力较低,但将该方法与其他已建立的分型方法(如RAPD和ERIC-PCR)结合使用,可能有助于对气单胞菌分离株进行可靠分型或鉴定具有致病潜力的新蛋白。