RAPD- and ERIC-Based Typing of Clinical and Environmental Pseudomonas aeruginosa Isolates

• Published in: Journal of AOAC International 2017-03, Vol.100 (2), p.532-536
• Main Authors: Auda, Ibtesam Ghadban, Al-Kadmy, Israa M S, Kareem, Sawsan Mohammed, Lafta, Aliaa Khyuon, A'Affus, Mustafa Hussein Obeid, Khit, Ibrahim Abd Aloahd, Al Kheraif, Abdulaziz Abdullah, Divakar, Darshan Devang, Ramakrishnaiah, Ravikumar
• Format: Article
• Language: English
• Subjects: Bacterial Typing Techniques; Genetic aspects; Genetic screening; Genotype; Health aspects; Humans; Identification and classification; Methods; Microbiological research; Pseudomonas aeruginosa; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - isolation & purification; Random Amplified Polymorphic DNA Technique
• ISSN: 1060-3271; 1944-7922
• DOI: 10.5740/jaoacint.16-0267

Pseudomonas aeruginosa is a major cause of nosocomial infection in children and adults, resulting in significant morbidity and mortality due to its ability to acquire drug resistance. The ability of P. aeruginosa in the environment to cause infection in individuals has been reported previously; henceforth, surveillance of the emergence and transmission of P. aeruginosa strains among patients is important for infection control in a clinical setup. Various gene-typing methods have been used for epidemiological typing of P. aeruginosa isolates for the purpose of surveillance. In this work, the suitability and comparability of two typing methods, enterobacterial repetitive intergenic consensus (ERIC)-PCR and random amplification of polymorphic DNA (RAPD)-PCR fingerprinting, were studied to characterize P. aeruginosa strains isolated from clinical and environmental sources. Forty-four clinical and environmental bacterial isolates of P. aeruginosa were collected between October 2015 and January 2016. DNA extraction, ERIC-PCR and RAPD-PCR, agarose gel electrophoresis, and phylogenetic analyses were carried using the unweighted pair-group method with mean. RAPD typing revealed less clonality among clinical isolates, whereas the ERIC method showed greater similarity in comparison with RAPD. Environmental isolates, however, showed greater similarity using RAPD compared with ERIC typing. With only a few exceptions, most clinical isolates were distinct from environmental isolates, irrespective of the typing method. In conclusion, both the RAPD and ERIC typing methods proved to be good tools in understanding clonal diversity. The results also suggest that there is no relationship between clinical and environmental isolates. The absence of clonality among the clinical isolates may indicate that most P. aeruginosa infection cases could be endemic and not epidemic and that endemic infections may be due to nonclonal strains of P. aeruginosa.