Molecular Analysis of the Isolates of Acinetobacter baumannii isolated from Tehran Hospitals Using ERIC-PCR Method

Abstract

Background and Objective:Acinetobacter is a genus of non-fermenting Gram-negative cocci or coccobacilli, which have low nutritional requirements for growth and can survive for a long time in adverse conditions, on dry surfaces, and also in aqueous environments. The importance of the members of Acinetobacter genus as pathogens involved in nosocomial infections, is increasing. Acinetobacter baumannii is the most common species involved in a broad spectrum of nosocomial infections, including pneumonia, bacteremia, surgical wound infections, urinary tract infections, and meningitis. In this study, enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR) technique was used for analysis and molecular typing of Acinetobacter strains, which has high discrimination power compared to phenotypic markers. Methods:In the present study, a total of 40 A. baumanniies strains were isolated from patients hospitalized in Tehran hospitals. After identification and confirmation of the isolates by serotyping and biochemical tests, a single colony of each isolate was cultured on liquid LB medium, and after DNA extraction, PCR was performed. After electrophoresis of PCR product, gel images were stored electronically for analysis and comparison of the isolates. Results:In this study, 40 strains of A. baumannii were analyzed by ERIC-PCR method, of which 29 strains were typed into 10 groups and 11 other strains had no PCR bands or had a band that could not be assigned to any of the above groups. Conclusion:In this study, it was found that A. baumanniie strains could be typed using repetitive sequences. This extent of polymorphism shows that ERIC-PCR is a useful method for analysis of genetic variation of A. baumannii strains. High genetic variation of A. baumannii strains may be due to wide geographical distribution of this species in Iran

Distribution and Genotyping of Aquatic Acinetobacter baumannii Strains Isolated from the Puzi River and Its Tributaries Near Areas of Livestock Farming

“…Further, cluster II contained isolates from the other Puzi River area (p-value = 90%) ( Figure 3). Maleki et al (2016) found that the diversity of genetic patterns of A. baumannii that were observed by ERIC-PCR analysis was due to the wide distribution in hospitals [2]. Here, we postulate that this high diversity is due to the distributions of sampling sites with different sources.…”

“…Molecular genotyping methods, including plasmid profiling, ribotyping, PFGE, MLST, and PCR-based typing methods, have been evaluated as potential methods to characterise A. baumannii isolates [6]. Despite MLST and PFGE being highly discriminative genotyping methods, PCR-based DNA fingerprinting techniques have advantages of performance ease and economic viability [2,6,12]. To date, there have been many studies describing different PCR-based methods to type MDR-AB [47][48][49].…”

“…Acinetobacter baumannii is a Gram-negative, non-fermenting, aerobic bacterium [1][2][3] which is considered to be ubiquitous as it can be recovered from various environments, including soil or surface water [4][5][6][7]. It is also an important health care-associated pathogen, which mostly causes opportunistic infections in immunocompromised individuals [6].…”

“…In the treatment of various A. baumannii infections, a reliable method for identification and characterization of the strains is necessary. Numerous methods have been developed for analysis and molecular typing of A. baumannii, such as plasmid analysis, ribotyping, multilocus sequence genotyping (MLST), pulsed-field gel electrophoresis (PFGE), and several other sequence-based polymerase chain reaction (PCR) typing techniques [2,4,6,[11][12][13]. Despite MLST and PFGE being highly discriminative genotyping methods, sequence-based DNA fingerprinting PCR techniques have advantages of performance ease and economic viability [2,12,14].…”

“…Numerous methods have been developed for analysis and molecular typing of A. baumannii, such as plasmid analysis, ribotyping, multilocus sequence genotyping (MLST), pulsed-field gel electrophoresis (PFGE), and several other sequence-based polymerase chain reaction (PCR) typing techniques [2,4,6,[11][12][13]. Despite MLST and PFGE being highly discriminative genotyping methods, sequence-based DNA fingerprinting PCR techniques have advantages of performance ease and economic viability [2,12,14]. Since the enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique has a sound discrimination index (over 94%) [12] and a relatively low cost for large-scale genotyping of A. baumannii when compared with other sequence-based PCR techniques, it has been widely adopted in the studies of environmental A. baumannii studies.…”