Determination of Antibiotic Resistance Profile and Frequency of

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Determination of Antibiotic Resistance Profile and Frequency of
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ƭŶºƗƶºƫŚƷźºƐƣƂƿřżºƟřƢƿźƏŻř MBLS ŶǀƫƺţŵřźĭƾŤƳŚſ
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ŻřŶºƿŵźĭƆŴƄºƯƮƴěƾưƿřĨƀƿŵŚŝƶƀƿŚƤƯŹŵEDTA
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ŚºŝƺºĮƫřDNAÏxlTaq DNA polymerase Îunit
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Gene Atlas ÐÏÏ system, (ASTEC, źƬĪƿŚºſƺƯźţ
ŶƃƭŚŬƳřźƿŻJapan
ATCC ÏÔÕÒÐřŻƺºƴƿĥƹźŗōŽŚƳƺƯƹŵƺºſŵŹřŶƳŚŤºſřƽƶƿƺſ
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blaIMP, blaSPM, blaSIMƽŚºƷƱĥƪƯŚů
ƽŚƷƶƿƺſŶƿŵźĭ
ƾºſŚƴƃ śƹźºƨǀƯ ƂºŴŝŻřƵŶƃƶǀƸţŢŞŨƯƩźŤƴĩƱřƺƴƗƶŝ
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ƽŶǀţƺŘƬĩƺƳƾƫřƺţ
blaIMP-F
blaIMP-R
blaSPM-F
blaSPM-R
blaSIM-F
blaSIM-R
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5'-CCAAACCACTACGTTATCT-3'
5'-AAAATCTGGGTACGCAAACG-3'
5'-ACATTATCCGCTGGTACAGG-3'
5'-TACAAGGGATTCGGCATCG-3'
5'-TAATGGCCTGTTCCCATGTG-3'
ƩƺƈŰƯƵŻřŶƳř
188 bp
271 bp
570 bp
ŚƷƶŤƟŚ
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ƵŶºƃ ƽŹƹōƖưū ƽřŻƺƴƿĥƹźŗōŽŚƳƺƯƹŵƺſ ƶƫƹżƿř ÎÏÍ Żř
ƶŤºƃŹƹŵƱŶºƃŻŚºŝƽƶºƬůźƯƶºƤǀƣŵê šŶºƯƶŝŵřźĭƾŤƳŚſ
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Źŵ(Annealing Step)ŚºƷźưƿřźě ƩŚƈţř ƽƶƬůźƯ
ƶǀƳŚŧÐÍ
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ƽƶºūŹŵÔÏŹŵ(Extension Step)ƝŶºƷ ƽƶŤºƃŹ ƱŶºƃ
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ƾƿŚºƸƳƱŶºƃ ƪºƿƺƏ ƽƶºƬůźƯ ƹ ƶǀƳŚŧ ÒÍ šŶƯ ƶŝ ŵřźĭƾŤƳŚſ
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.ƶƤǀƣŵ ÕšŶƯƶŝ
ƍƺŝźƯƶƳƺưƳŶƇŹŵÑÏÒÒΝƾſŹźŝŵŹƺƯƾƴǀƫŚŝƽŚƷƶƳƺưƳ
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ƱŚƄºƳŶƇŹŵÏÎÓƶƫƹżƿřÏÓŹŵƲǀſŚĪǀƯōƹŶƇŹŵÑÐÐ
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ŶºƇŹŵ ÏÖÒ ƹŶºƇŹŵ ÐÑÑ ƲǀĮƳŚǀƯŹƺƏƶŝŶƿŻƺĮǀƬĭƺƴǀƯō
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ƿřŢƯƹŚƤƯ ŶƇŹŵÎŹřŵƺưƳ
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ƽŚºƷƶºƳƺưƳƶŝƍƺŝźƯŶƇŹŵÕӝƾƀƠƴţƽŚƷƶƳƺưƳƶŝƍƺŝźƯ
ŶºƫƺƯ ƽřŻƺºƴƿĥƹźŗōŽŚƳƺƯƹŵƺºſ ƶºƫƹżƿř ÐÒ ŻřŶƳŵƺŝ Ɓƺĭ
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ƹŵ
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PCRšLJƺƈŰƯŻŹƺƟƹźŤĪƫřÎƪĪƃ
ƶƿƺſ
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ÏĨƷŚģblaIMP ƱĥŶūřƹŢŞŨƯƩźŤƴĩƽƶƳƺưƳ
ÎĨƷŚģÎÍÍbpƾƫƺĪƫƺƯƱŻƹźĩŚƯM
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Multiplex PCRƩƺƈŰƯŻŹƺƟƹźŤĪƫřÏƪĪƃ
.blaIMPƱĥŶūřƹƾ
ƴǀƫŚŝƽ
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ƽƶƳƺưƳ
ÎĨƷŚģÎÍÍbp
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ƶƳƺưƳ
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ƶƳƺưƳ
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ƶƳƺưƳ
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ÓĨƷŚģblaSIMƱĥŶūřƹƾƴǀƫŚŝ
ƶƳƺưƳ
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ÒĨƷŚģ
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ŦŰŝ
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ÎÐÖÐƱŚŝōƹźƸƯÖÐƽƵŹŚưƃ
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7-
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Determination of Antibiotic Resistance Profile and Frequency ofMetalloBeta- Lactamases in Pseudomonas Aeruginosa Isolates
Hemmati F1, Sourouri Zanjani R2, Haghi F1, Zeighami H1
1
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Dept. of Microbiology, Zanjan University of Medical Sciences, Zanjan, Iran
2
Dept. of Microbiology, School of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran
Corresponding Author: Sourouri Zanjani R, Dept. of Microbiology, School of Medicine, Baqiyatallah University of
Medical Sciences, Tehran, Iran
E-mail: [email protected]
Received: 29 Sep 2013
Accepted: 2 Mar 2014
Background and Objective: Pseudomonas aeruginosa is an opportunistic pathogen and one of the most
frequent pathogens in nosocomial infections. This bacterium shows high resistance to majority of antibiotics
including â-lactams.The aim of this study was to investigate the pattern of antimicrobial resistance and
evaluation of IMP, SPM and SIM Metallo- Beta- Lactamase producing Pseudomonas aeruginosa, in clinical
samples isolated from Zanjan hospitals, by phenotypic and PCR methods.
Materials and Methods: In this cross-sectional study, 120 Pseudomonas aeruginosa isolates were collected
from the various clinical specimens in Zanjan hospitals from 2013 to 2014. After identification of isolates
using biochemical tests, the antibiotic susceptibility test (Kirby-Baur method) was done according to CLSI
advice against 7 antibiotics. The Combined Disk method was then carried out for detection of MBLs and the
blaIMP, blaSPM and blaSIM genes were determined by a PCR method.
Results: In this study, Cefotaxime and Amikacin with 43.3% and 21.6% showed the highest and lowest
resistance against isolates, respectively. From a total of 120 isolates, 35 (29.2%) strains were imipenem
resistant and metallo- beta-lactamase producer. From 35 MBL producing isolates, 28 (80%) strains, 20
(57.1%) strains and 5 (14.3%) strains carried blaIMP, blaSPM and blaSIM genes, respectively and 1 (2.8%)
isolate carried all blaIMP, blaSPM and blaSIM genes.
Conclusion: According to the results, high prevalence of resistance to aztreonam and imipenem can be a
warning against treatment process and distribution of resistance to other strains.
Keywords: Antibiotic resistance, Mtallo-Beta-Lactamase, Pseudomonas aeruginosa
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