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Abstract

Background: Multiple-drug resistant Acinetobacter have widely spread in the last decades imposing a serious nosocomial source of infection. Nevertheless, little knowledge was gaimed on tracing the development of antibiotic resistance in Acinetobacter species. Objectives: Explore Acinetobacter spp. via antimicrobial susceptibility, plasmid profiles, and random amplified polymorphism DNA polymerase chain reaction (RAPD-PCR) typing. Methods: One hundred twelve Acinetobacter isolates (including 66 A. baumannii and 46 non-Acinetobacter baumannii strains) were obtained from three university hospitals. The source of infection of these isolates included blood, urine, wound, and respiratory tract. Their susceptibilities to 17 antibiotics were tested and then all Acinetobacter isolates were typed by plasmid analysis and RAPD-PCR method. Results: A. baumannii isolates revealed nine different patterns of antibiotic resistance. Of those, non- A. baumannii, were associated with plasmid and RAPD-PCR typings (p <0.05). A. baumannii was more resistant to multiple antibiotics than non-A. baumannii (p <0.05). Seven different plasmid profiles were observed among 112 Acinetobacter isolates. Plasmids were found in 107 (95.5%) of the 112 isolates. Unlike in RAPD-PCR typing, there was no difference between the type of Acinetobacter, A. or non-A. baumannii strains and plasmid profiles (p >0.05). By RAPD-PCR, six profiles were found for each A. and non-A. baumannii strains. The pattern 6 was the most common pattern among the isolates. Both plasmid and RAPD-PCR typing showed no association between plasmid profiling and site of infection (p >0.05). Conclusion: There is a wide spread of multi-drug resistant Acinetobacter spp., particularly A. baumannii, in the Middle East region that can be traced efficiently by plasmid and genotyping typing of Acinetobacter. More care should be taken for tracing the development of antimicrobial resistance of Acinetobacter using precise molecular typing techniques.

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diseases Society of America. Clin Infect Dis 2010;50:291-322. 12. Park BJ, Wannemuehler KA, Marston B J, Govender N, Pappas PG, Chiller TM. Estimation of the current global burden of cryptococcal meningitis among persons living with HIV/AIDS. AIDS 2009;23:525-30. 13. Mlinarić-Missoni E, Hagen F, Chew WHM, Važić-Babić V, Boekhout T, Begovac J. In vitro antifungal susceptibilities and molecular typing of sequentially isolated clinical Cryptococcus neoformans strains from Croatia. J Med Microbiol 2011;60:1487-95. 14. Murray PR, Rosenthal KS, Pfaller MA. Medical

, Paterson DL, Rice LB, Stelling J, Struelens MJ, Vatopoulos A, Weber JT, Monnet DL. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 2012;18:268-81. doi: 10.1111/j.1469-0691.2011.03570 25. Célia Maria Carvalho Pereira Araújo Romão, Faria YN, Pereira LR, Asensi MD. Susceptibility of clinical isolates of multiresistant Pseudomonas aeruginosa to a hospital disinfectant and molecular typing. Mem Inst Oswaldo Cruz 2005;100:541-8. doi

mechanisms and specific resistances of clinical and/or epidemiological importance.Version 1.0, 2013. http://www.eucast.org . 19. Ribot, E.M., Fair, M.A., Gautom, R., Cameron, D.N., Hunter, S.B., Swaminathan, B., Barrett, T.J. (2006). Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne Pathog Dis. 3(1): 59-67. https://doi.org/10.1089/fpd.2006.3.59 PMid:16602980 20. Caprioli, A., Maugliani, A., Michelacci, V., Morabito, S. (2014). Molecular typing of Verocytotoxin

first year of life. Acta Vet Scand. 49, 22. http://dx.doi.org/10.1186/1751-0147-49-22 PMid:17848186; PMCid:PMC2040143 12. Beck, R., Sprong, H., Bata, I., Lucinger, S., Pozio, E., Cacció, S. M. (2011b). Prevalence and molecular typing of Giardia spp. in captive mammals at the ZOO of Zagreb, Croatia. Vet Parasitol. 175, 40-46. http://dx.doi.org/10.1016/j.vetpar.2010.09.026 PMid:20970259

Reservoirs of drug resistant bacterial genomes and extrachromosomal DNA segments are a growing problem and cause emergence of new multidrug resistant (MDR) strains [ 1 ]. Antibiotic resistance of Klebsiella infections are causing increasing morbidity and mortality, and an increase in health care costs worldwide. In epidemiological research, not only phenotypical analysis, but also genotypical analysis is conducted by using various molecular typing methods such as plasmid profiling, ribotyping, and polymerase chain reaction (PCR) to find genetic relationships

., Dondo, A., Tagliabue, S., Bonora, S., Zanardi, G., Pacciarini, M.L. (2009). Molecular typing of Mycobacterium bovis strains isolated in Italy from 2000 to 2006 and evaluation of variablenumber- tandem-repeats for a geographic optimized genotyping. J Clin Microbiol. 47 (3): 636-644. https://doi.org/10.1128/JCM.01192-08 PMid:19144792 PMCid:PMC2650904 44. Prodinger, W.M., Brandstätter, A., Naumann, L., Pacciarini, M., Kubica, T., Boschiroli, M.L., Aranaz, A., Nagy, G., Cvetnic, Z., Ocepek, M., Skrypnyk, A., Erler, W., Niemann, S., Pavlik, I., Moser, I. (2005

Miguel, M.J., Muñoz, P.M., Blasco, J.M., Jacques, I., Grayon, M., Cloeckaert, A., Ferreira, A.C., Cardoso, R., Corrêa de Sá, M.I., Walravens, K., Albert, D., Garin-Bastuji, B. (2008). Evaluation of a multiplex PCR assay (bruce-ladder) for molecular typing of all Brucella species, including the vaccine strains. J. Clin. Microbiol.,46, 3484-3487. http://dx.doi.org/10.1128/JCM.00837-08 PMid:18716225 PMCid:PMC2566117 20. García-Yoldi, D., Marín, C.M., de Miguel, M.J., Muñoz, P.M., Vizmanos, J.L., López-Goñi, I. (2006). Multiplex PCR assay for the identification and