Molecular Detection of Carbapenem Resistance in Acinetobacter Baumannii Isolated From Patients in Khorramabad City, Iran

(1)

See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/333175528

Molecular Detection of Carbapenem Resistance in Acinetobacter Baumannii Isolated From Patients in Khorramabad City, Iran

Article · May 2019

DOI: 10.2174/1871526519666190517124314

CITATIONS

0

READS

34 6 authors, including:

Somayeh delfani Lorestan University 16PUBLICATIONS 208CITATIONS

SEE PROFILE

Faranak Rezaei Lorestan University 15PUBLICATIONS 168CITATIONS

SEE PROFILE

All content following this page was uploaded by Faranak Rezaei on 26 April 2020.

The user has requested enhancement of the downloaded file.

(2)

Infectious Disorders - Drug Targets, 2019, 19, 1-7 1

RESEARCH ARTICLE

Molecular Detection of Carbapenem Resistance in Acinetobacter Bauman- nii Isolated From Patients in Khorramabad City, Iran

Zeinab Babaie

¹

, Somayeh Delfani

^1,2*

, Faranak Rezaei

^1,2

, Fatemeh Norolahi

³

, Somayeh Mahdian

⁴

and Pegah Shakib

²

1Department of Microbiology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran, ²Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran,³Department of Mi- crobiology, School of Medicine, Pasteur Institute University of Medical Sciences, Tehran, Iran,⁴Department of Micro- biology, School of Medicine, Ilam University of Medical Sciences, Ilam, Iran

A R T I C L E H I S T O R Y Received: November 22, 2018 Revised: February 04, 2019 Accepted: February 04, 2019

DOI:

10.2174/1871526519666190206210808

Abstract: Background: Acinetobacter baumannii is an opportunistic pathogen, which causes a wide range of infections in hospitals, especially in intensive care units. Nowadays, due to the high resistance of Acinetobacter bumanni to antibiotics, this study, in addition to the phenotypic and genotypic investigations of drug resistance, focused on determining the molecular types of Acine- tobacter baumannii isolated from patients in Khorramabad city by the pulsed-field gel electrophoresis (PFGE) method

Materials and Methods: In this cross-sectional study, 50 samples of Acinetobacter baumannii were collected from educational hospitals in Khorramabad city, Iran, from January to August 2015. They were identified in the laboratory using biochemical tests and culture methods. After determining the drug resistance pattern by the disc diffusion method and percentage of resistance genes to carbapenems, Acinetobacter baumannii isolates were analyzed using the PFGE method using the Apa1 enzyme.

Results: The highest antibiotic resistance observed for Acinetobacter baumannii strains was against ampicillin-sulbactam (100%) and aztreonam (98%). The highest sensitivity was to po- lymixin B (100%) and colistin (94%), and also to the OXA-51-like gene present in all samples.

The OXA-23-like gene was positive in 44 (88%) samples. PFGE results showed that Acinetobac- terbaumannii strains had 33 different pulsotype patterns, of which 27 patterns had more than one strain and 23 had only one strain.

Conclusion: Due to the high resistance of Acinetobacter baumannii and its ease of spread and its ability to transfer resistance genes, resistance control methods should be used in the disinfection of hospital areas. Hospital staff should observe hygiene standards and there should also be a re- duction in antibiotic use.

Keywords: Acinetobacterbaumannii, antibiotic resistance, pulsed-field gel electrophoresis (PFGE).

1. INTRODUCTION

Acinetobacter baumannii is a coccobacillus, gram- negative, negative oxidase, obligate aerobic, non-fermented, and non-motile [1]. With few requirements for growth, this bacterium can survive long-term in adverse, dry and aquatic environmental conditions [2]. Acinetobacter baumannii is widely distributed in the hospital environment and is easily transmitted among patients, especially patients admitted to the intensive care unit [3]. A wide range of infections, including urinary tract infections, meningitis, bacteremia,

*Address correspondence to this author at the SomayehDelfani, Department of Microbiology, School of Medicine, Lorestan University of Medical Sci- ences, Khorramabad, Iran; E-mail: [email protected]

wound infections, skin and soft tissue infections, and respiratory infections are due to Acinetobacter baumannii activity [1, 4, 5].

Nowadays, Acinetobacter baumannii is resistant to most antibiotics, including aminoglycosides, carbapenems, cephalosporins and beta-lactams [6-8]. The factor that strengthens this resistance system is its intrinsic ability to survive long- term in all hospital settings, which has led to an increase of the prevalence of this bacterium in hospitals [9, 10].

One of the drugs used worldwide to treat this bacterium infection is the beta-lactam family of antibiotics [11, 12]. But as mentioned, these bacteria are resistant to beta-lactams, due to genetic elements. The most common genes associated with resistance to beta-lactam resistant isolates are

(3)

2 Infectious Disorders - Drug Targets, 2019, Vol. 19, No. 00 Babaie et al.

ampCcephalosporinase genes, OXA type carbapenems, metallobetalactamas, tracheal pumps, and introns [13-16].

Currently, carbapenems are the preferred antibiotics against multi-drug resistant Acinetobacter infections. However, resistance to carbapenems has been a major concern for public health, and there are limited therapeutic options to prevent the horizontal transfer of resistant genes and the inherent differential expression of the genes.

There are many methods for molecular typing of bacteria.

Genotyping techniques are used in the Acinetobacter bau- mannii assimilation. Some of these methods are comprised of Pulsed Field Gel Electrophoresis (PFGE), Ribotyping, Random Amplification of Polymorphic DNA, Amplified fragment length polymorphism, and Repetitive element sequence-based PCR(REP-PCR) [17]. In the meantime, the PFGE technique is considered as the gold standard method for determining the species of Acinetobacter bumannii [10, 18]. Although this method is complicated, its equipment is currently not only found in reference laboratories, but also in some of the advanced laboratories in hospitals. Typically, limited Apa1 and Sma1 enzymes are used to cut chromo- some DNA. Then, chromosomal portions created by electrophoresis are separated from each other, and fingerprinted graphs are compared either by eye or using computer pro- grams [19].

Using this method, no strain of Acinetobacter baumannii had previously been isolated in Khorramabad. This study was conducted to analyze the phenotypic and genotypic resistance of this bacterium, with the aim of determining the molecular types of Acinetobacter baumannii isolated from patients in Khorramabad city by the PFGE method.

2. MATERIALS AND METHODS 2.1. Bacterial Isolates

For this cross-sectional study, Acinetobacter baumannii isolates were collected from educational hospitals in Khor- ramabad, Iran, from January to August 2015. In the laboratory, after culture of the isolates on a MacConkey agar me-

dium, Acinetobacter baumannii isolates were identified by standard biochemical tests.

2.2. Antimicrobial Susceptibility Pattern

In order to investigate the resistance of Acinetobacter baumannii, disc diffusion tests were performed for 13 different antibiotics from MAST UK, including cefotaxime(30 µg), colistin(10 µg) , meropenem(10 µg), polymexin B(300 µg), aztreonam(30 µg), tetracycline(30 µg), cefepime(30 µg),trimethoprim-sulfamethoxazole (1.25/23.75 µg), ampicillin-sulbactam(10/10 µg), amikacin (10 µg), tobramycin(10 µg), ceftriaxon (30 µg), and ciprofloxacin (5 µg), using the Antibiotic disc diffusion method. For the purpose of quality control of the experiments, the standard strain of Es- cherichia coli ATCC 25922 was used.

2.3. Polymerase Chain Reaction (PCR) Detection of Car- bapenemase Genes

To perform the PCR, 24-hour cultures of the bacteria were collected from a plate containing Mueller-Hinton agar in a loop size, then made up to a 200 ml sterilized water sus- pension and boiled for 10 minutes. After centrifugation, the conducted supernatant was used as a template DNA for PCR.

Duplication was performed using a thermocycler machine.

Traceable genes included bla_OXA-51, bla_OXA-58, bla_OXA-23, blaOXA-24, and ISA-ba1 genes. The primers which were used in this study are written down in Table 1, as well as the reaction conditions as prescribed on the pertinent documentation.

2.4. Pulsed Field Gel Electrophoresis (PFGE)

To perform PFGE, isolates of patients were cultured on TSA agar for 18 to 24 hours and incubated at 37°C and used to prepare the DNA and the plug according to the standard protocol. To prepare the plug, 400 µl of each microbial sus- pension was poured into a microtube and 400 µL of LMP agarose was added (agarose dissolved in TE buffer). Also, for each sample, 20mg / ml were added to each micro tube and placed into the plug molding unit.

The samples were kept at room temperature for 5-15 minutes andthenkeptin the refrigerator for 10 minutes. Af- Table 1. Sequence of primers for resistance to carbapenems genes and expected PCR product length.

Primer Name Primer Sequence Fragment Length(bp) Reference

BlaOXA-51-like

5´-TAA TGC TTT GAT CGG CCT TG-3´

5´-TGG ATT GCA CTT CAT CTT GG-3´ 353 [20]

BlaOXA -23- like

5´-GAT CGG ATT GGA GAA CCA GA-3´

5´- ATT TCT GAC CGC ATT TCC AT-3´ 501 [20]

BlaOXA -24- like

5´-GGT TAG TTG GCC CCC TTA AA-3´

5´AGT TGA GCG AAA AGG GGA TT-3´ 246 [20]

BlaOXA-58-like

5´-AAG TAT TGG GGC TTGTGC TG-3´

5´-CCC CTC TGC GCT CTA CATAC-3´ 559 [20]

ISAba1 5´-CATTGGCATTAAACTGAGGAGAAA-3´

5´ -TTGGAAATGGGGAAAACGAA-3´ 451 (21)

(4)

Table 2. Percentage of phenotypic resistance of different antibiotics of Acinetobacter baumannii.

Antibiotic Resistant Strains N (%) Intermediate Strains N (%) Sensitive Strains N (%)

Cefotaxime 48(96%) 1(2%) 1(2%)

Colistin 3(6%) 0(0%) 47(94%)

Meropenem 45(90%) 2(4%) 3(6%)

Polymyxin B 0(0%) 0(0%) 50(100%)

Aztreonam 49(98%) 0(0%) 1(2%)

Tetracycline 43(86%) 3(6%) 4(8%)

Cefepime 45(90%) 1(2%) 4(8%)

cotrimoxazole 43(86%) 3(6%) 4(8%)

Ampicillin-sulbactam 50(100%) 0(0%) 0(0%)

Amikacin 17(34%) 17(34%) 16(32%)

Tobramycin 43(86%) 4(8%) 3(6%)

Ceftriaxone 48(96%) 1(2%) 1(2%)

Ciprofloxacin 47(94%) 1(2%) 2(4%)

Fig. (1). Prevalence of antibiotics resistance among Acinetobacter baumannii isolates with 95% confidenceinterval.

ter transferring these plugs into the test tube, lysis buffer was applied for 1.5 to 2 hours inside the shaker incubator at 55°C. Then each plug was washed twice with distilled water and 4 times with TE buffer. At the last wash, after emptying the TE buffer into the tube containing the plug, fresh buffer was added and the samples were placed at a temperature of 4-8°C. In the next step, for enzyme digestion and endonucle- ase genomic DNA fragmentation, Apa1 enzyme was used according to the protocol, the enzyme digestion was as fol- lows. In the first step, the plugs were first mixed with 180cc distilled water and 20 µl of enzyme buffer (10X) and incubated microtubes for 15 minutes at room temperature, and

again the plugs were transferred to another microtube, and were mixed with 175 µl of distilled water, 20 µl, 10X buffer and 5µl of the Apa1 enzyme and incubated for 5 hours at 25°C in a shaker incubator. Electrophoresis was performed in 1% agarose gel in Chef Drii in TBE 0.5X buffer with an initial switching program of 1 minute and 35 seconds, with a final switching time of 35 seconds, and a runtime 19 hours at 6 volts/cm. In the next step, the gel was stained in Ethidium bromide, washed with distilled water, and then analysed in a gel doc machine. Finally, the dendrogram pattern was determined and the relation of the samples clonal to each other was determined using Gel comparison software.

(5)

2.5. Statistical Analysis

Data were collected on standard forms, entered into an Excel database and analyzed by using the SPSS software version 20.0.

3. RESULTS 3.1. Clinical Isolates

Fifty isolates were identified as Acinetobacter bauman- nii, by standard biochemical tests.

3.2. Antimicrobial Resistance of Acinetobacter baumannii strains

The highest antibiotic resistance of Acinetobacter bau- mannii strains was respectively ampicillin sulbactam (100%), aztreonam (98%), cefotaxime and ceftriaxone (96%), ciprofloxacin (94%), cefepime and meropenem (90%), tetracycline, tobramramycin and Trimethoprim- Sulfamethoxazole (86%) and amikacin (44%), the highest antibiotic susceptibility was polymyxin B (100%) and

colistin (94%), respectively. The results of the phenotypic resistance study of Acinetobacter baumannii strains are shown in Table 2 and Fig. (1).

3.3. PCR Amplification for Detection of blaOXA-51, blaOXA- 58, blaOXA-23, blaOXA-24, and ISA-ba1 genes

Acinetobacter baumannii distribution resistance genes in clinical strains are demonstrated in Table 3. All Acinetobac- ter baumannii strains contained the blaOXA-51-like gene.

3.3. Results of PFGE Method

In this study, all 50 strains of PFGE were typed. The results were analyzed using Gel comparison software. Based on an 80% cut off, 34 different patterns of different pulsating patterns were observed. Of the 34 patterns, 27 patterns contained more than one Acinetobacter baumannii strain, and 23 patterns had only one strain.

In this study, the highest frequency was related to patterns of 3,4,5,6,7, with 5 strains, and the lowest frequency of Table 3. Percent of resistance genes in Acinetobacter baumannii clinical strains.

Genes Resistant Strains N (%)

blaOXA-23 like 44(88%)

ISAba1 43(86%)

Fig. (2). Acinetobacter baumannii strain dendrogram with Gel compare software.

(6)

strains 1,2,8,9,10,12,14,15,17,18,20, 21,22,24,25,27,30,35, 36,37,38,45,48 with one strain. The dual strains included [46 and 47], [39 and 50], [43 and 49], [23 and 42], [26 and 28], [34 and 40], [41, 44] and [31 and 29] and triple strains [13, 16 and 19 and 11, 32 and 33]. The results of the PFGE study of strains are shown in Fig. (2).

4. DISCUSSION

Acinetobacter baumannii has been responsible for the spread of many hospital infections in recent decades. The bacterium is scattered throughout the world and causes many problems in humans [1, 22-25]. Previous studies have shown that front-line treatment for Acinetobacter baumannii infections includes amikacin, carbapenems, ceftazidime and quinolones. Among these, resistant strains to carbapenems have turned out to be a huge problem in the world, and are considered as a worldwide problem, since the treatment of these strains is very difficult [26-28]. Unfortunately, no data is available on the antibiotic resistance of Khorramabad city isolates of Acinetobacterbaumannii or on the spreading of genes involved in resistance to carbapenems.

In our study, all Acinetobacter bumanniistrains were resistant to Ampicillin Sulbactam, which was similar to the findings of Wang et al., 2003 [29]. The findings of this study showed that the resistance to meropenem (a member of the carbapenem family) was about 90%, contrary to the Hujer study in 2003, which reported resistance to carbapenems of only 20% [30]. This contrast is related to the difference in antibiotic use in different regions.

In the study of Acinetobacter baumannii antibiotic resistance by Simhon, the resistance to imipenem increased from 1.9% in 1990 to 35.9% in 2000 [31]. Similarly, studies such as TalebiTaher in Tehran in 2010-2011 (percentage resistance to imipenem was 97%), Zarrilli and Szejbach in 2013 (94.99%), indicate that resistance to carbapenems is increasing worldwide [32-34]. This study is similar to the study of Seng Stock and colleagues in the United States, of which more than 98% of the groups were resistant to azotronam [35]. The results of this study, and its comparison with similar studies indicate that resistance to Acinetobacter bauman- nii is constantly changing, and can cause serious concerns in the field of treatment. Differences in the clinical samples studied, the timing of the study and treatment strategies in each geographic region may be responsible for the differences in the results of different studies.

The results of this study, as reported by Saeed Shoja (2016) in Ahvaz, BagheriJoushgani (2016) in Kashan and Azimi in Tehran (2013), were 100% resistant to the bla_OXA-51 gene [36-38]. In a study conducted by Shahchegrahi et al. in Tehran in 2011 on 100 samples, the percentage of resistance to the bla_OXA-51genome was 94% and bla_OXA-2384% [39]. In the study by Karmostaji in Tehran in 2013, the distributions of bla_OXA-51, bla_OXA-23, bla_OXA-24, bla_OXA-58resistance genes were 93.3%, 81.3%, 8.1%, and 0.81%, respectively [40]. In the study by Azizi in 2015, in 65 samples, bla_OXA-51and blaOXA-23 genes were found in 100% of samples, blaOXA-24 in 29 samples (44%) and bla_OXA-58in no samples [41]. In a study conducted by BagheriJoshagani in 2016, the percentage of resistance to bla_OXA-23and blaOXA-24 was 90%, ISAba1,

and blaOXA-51 was 100% and bla_OXA-58were reported as zero samples [36]. In a study conducted by NajarPiraye in Bandar Abbas, Iran in 2014, 78.9% of the samples had OXA-23 resistance gene, 8.8% had blaOXA-24 gene and 1.7% had blaOXA- 58 gene [42]. In the study by RabahBakour in Algeria in 2015, bla_OXA-51, bla_OXA-23and bla_OXA-24carbapenemase genes were detected in 100%, 67.02% and 20.21% of isolates, respectively [43]. In a study conducted by We, A Gebreys in the USA in 2009 on 83 samples, bla_OXA-23 in 13% and IS- Aba1 in 79.5% of clinical isolates was reported [44].The highest existence of resistance was in the bla_OXA-51, bla_OXA-23 and ISAba1 genes, and the lowest percentage in the bla_OXA-58 gene. These are the results of various studies, which are in agreement with our findings. The difference in the prevalence of carbapenemase genes in different regions is due to differences in the pattern of use of beta-lactam and carbapenem antibiotics; so increasing the resistance to beta- lactams, due to the presence of plasmids, also causes resistance to other antibiotics.

In a study by Lin MF et al. in 2011 in China, 11 pulsotypes were obtained from 23 Acinetobacter baumannii isolates by the PFGE technique, which named them A to K respectively [45]. Zarrili, 2004, in Italy, in a genotypic analysis of the Acinetobacter baumannii strains by PFGE from 131 patients revealed 9 distinct PFGE patterns, of which seven had only one strain [46]. Shahchegrahi et al. 2010, in Iran, acquired six types of 29 analyzed isolates, with type A being the most common type that included 19 isolates. The types named as B and C each were four and three isolates respectively, and the rest of D to F each contained one isolate [39].

In the present study of 50 samples, 33 different pulsotype was found, with the highest number of samples in 3, 4,5,6,7, 5 strains. The genomic pattern obtained shows the different origin of the isolates. And as indicated in the results section, 23 pulsotypes had one strain and 11 pulsotypes had more than one strain.

CONCLUSION

Due to the high resistance of Acinetobacter baumannii and its wide distribution, as well as its ability to transfer resistance genes through moving genetic factors among impor- tant bacteria in hospitals, effective methods of hospital disinfection and sanitary observation are required. In addition, the use of broad-spectrum antibiotics should be reduced. Infor- mation from strain typing is, in fact, complementary to wide- spread epidemiological research, and its results, along with other phenotypic methods, such as antibiogram, is valuable in a limited time and space. As PFGE is directly related to the genetic sequencing of strains, it can be typed and repli- cated with good results and diagnostic power, and is nowadays a superior technique in tracing strain origin.

ETHICS APPROVAL AND CONSENT TO PARTICIPATE

Not applicable.

HUMAN AND ANIMAL RIGHTS

No Animals/Humans were used for studies that are base of this research.

(7)

CONSENT FOR PUBLICATION Not applicable.

AVAILABILITY OF DATA AND MATERIALS Not applicable.

FUNDING None.

CONFLICT OF INTEREST

The authors confirm that this article content has no conflict of interest.

ACKNOWLEDGEMENT

This study was carried out in the form of a research pro- ject approved by No A-10-1503-2, with the support of the research vice president of the Department of Microbiology and Razi Herbal Medicines Research Center of Lorestan University of Medical Sciences.

REFERENCES

[1] Peleg AY, Seifert H, Paterson DL. Acinetobacter baumannii:

emergence of a successful pathogen. Clinical microbiology reviews. 2008;21(3):538-82.

[2] Baumann P. Isolation of Acinetobacter from soil and water. Journal of bacteriology. 1968;96(1):39-42.

[3] Gaynes RP, Edwards JR, Jarvis WR, Culver DH, Tolson JS, Mar- tone WJ, et al. Nosocomial infections among neonates in high-risk nurseries in the United States. Pediatrics. 1996;98(3):357-61.

[4] Bergogne-Berezin E, Towner K. Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features.

Clinical microbiology reviews. 1996;9(2):148.

[5] Mak JK, Kim M-J, Pham J, Tapsall J, White PA. Antibiotic resistance determinants in nosocomial strains of multidrug-resistant Acinetobacter baumannii. Journal of antimicrobial chemotherapy.

2008;63(1):47-54.

[6] Boroumand M, Akhyani H, Sheikhvatan M, Yazdi SH, Saboorian R, Hashemi S, et al. Evaluation of antimicrobial resistance of Acinetobacter baumannii to Imipenem, Ciporofloxacin and Ceftaz- idime using E Test. Iranian Journal of Public Health.

2009;38(2):130-3.

[7] Seifert H, Baginski R, Schulze A, Pulverer G. Antimicrobial susceptibility of Acinetobacter species. Antimicrobial agents and chemotherapy. 1993;37(4):750-3.

[8] Shi Z-Y, Liu PY-F, Lau Y-J, Lin Y-H, Hu B-S, Shir J-M. Antimi- crobial susceptibility of clinical isolates of Acinetobacter baumannii. Diagnostic microbiology and infectious disease. 1996;24(2):81- 5.

[9] Karah N, Sundsfjord A, Towner K, Samuelsen Ø. Insights into the global molecular epidemiology of carbapenem non-susceptible clones of Acinetobacter baumannii. Drug Resistance Updates.

2012;15(4):237-47.

[10] Mo G, She D, Guan X, Cui J, Wang R, Chen L. Antimicrobial resistance and genotyping of Acinetobacter baumannii in ICU.

Zhonghuajie he hehu xi zazhi= Zhonghuajiehe he huxizazhi= Chi- nese journal of tuberculosis and respiratory diseases.

2010;33(9):656-9.

[11] Michalopoulos A, Falagas ME. Treatment of Acinetobacter infections. Expert opinion on pharmacotherapy. 2010;11(5):779-88.

[12] El Salabi A, Walsh TR, Chouchani C. Extended spectrum β- lactamases, carbapenemases and mobile genetic elements responsible for antibiotics resistance in Gram-negative bacteria. Critical reviews in microbiology. 2013;39(2):113-22.

[13] Magnet S, Courvalin P, Lambert T. Resistance-nodulation-cell division-type efflux pump involved in aminoglycoside resistance in

Acinetobacter baumannii strain BM4454. Antimicrobial agents and chemotherapy. 2001;45(12):3375-80.

[14] Nemec A, Dolzani L, Brisse S, van den Broek P, Dijkshoorn L.

Diversity of aminoglycoside-resistance genes and their association with class 1 integrons among strains of pan-European Acinetobac- ter baumannii clones. Journal of medical microbiology.

2004;53(12):1233-40.

[15] SADEGHIFARD NK, RANJBAR R, GHASEMI A, Pakzad I, ZAEIMI YJ, Zaheri A, et al. A study of antimicrobial resistance of acinetobacterbaumannii and non-acinetobacterbaumannii strains isolated from three hospitals in Tehran. 2006.

[16] Gu B, Tong M, Zhao W, Liu G, Ning M, Pan S, et al. Prevalence and characterization of class Iintegrons among Pseudomonas aeru- ginosa and Acinetobacter baumannii isolates from patients in Nan- jing, China. Journal of clinical microbiology. 2007;45(1):241-3.

[17] Liu PY-F, Wu W-L. Use of different PCR-based DNA fingerprint- ing techniques and pulsed-field gel electrophoresis to investigate the epidemiology of Acinetobacter calcoaceticus-Acinetobacter baumannii complex. Diagnostic microbiology and infectious disease. 1997;29(1):19-28.

[18] Nasonova E. Pulsed field gel electrophoresis: theory, instruments and applications. Tsitologiia. 2008;50(11):927-35.

[19] Dehbalaei MA, Najar-Peerayeh S, Behmanesh M, Taherikalani M.

Polyclonal Distribution of blaOXA-23 Gene Among Acinetobacter baumannii Isolated from Intensive Care Unit Patients in Tehran;

Pulsed-Field Gel Electrophoresis Analysis. Jundishapur Journal of Microbiology. 2018;11(1).

[20] Feizabadi M, Fathollahzadeh B, Taherikalani M, Rasoolinejad M, Sadeghifard N, Aligholi M, et al. Antimicrobial susceptibility patterns and distribution of blaOXA genes among Acinetobacter spp.

Isolated from patients at Tehran hospitals. Jpn J Infect Dis.

2008;61(4):274-8.

[21] Rafei R, Dabboussi F, Hamze M, Eveillard M, Lemarié C, Gaultier M-P, et al. Molecular analysis of Acinetobacter baumannii strains isolated in Lebanon using four different typing methods. PLoS One. 2014;9(12):e115969.

[22] Lee K, Yong D, Jeong SH, Chong Y. Multidrug-resistant Acineto- bacter spp.: increasingly problematic nosocomial pathogens. Yon- sei medical journal. 2011;52(6):879-91.

[23] Fournier PE, Richet H, Weinstein RA. The epidemiology and control of Acinetobacter baumannii in health care facilities. Clinical infectious diseases. 2006;42(5):692-9.

[24] Towner K. Acinetobacter: an old friend, but a new enemy. Journal of Hospital Infection. 2009;73(4):355-63.

[25] Wisplinghoff H, Schmitt R, Wöhrmann A, Stefanik D, Seifert H.

Resistance to disinfectants in epidemiologically defined clinical isolates of Acinetobacter baumannii. Journal of Hospital Infection.

2007;66(2):174-81.

[26] Prashanth K, Badrinath S. In vitro susceptibility pattern of Acine- tobacter species to commonly used cephalosporins, quinolones, and aminoglycosides. Indian journal of medical microbiology.

2004;22(2):97.

[27] Van Looveren M, Goossens H, Group AS. Antimicrobial resistance of Acinetobacter spp. in Europe. Clinical microbiology and infection. 2004;10(8):684-704.

[28] Ruiz M, Marti S, Fernandez-‐Cuenca F, Pascual A, Vila J. High prevalence of carbapenem-‐hydrolysingoxacillinases in epidemiologically related and unrelated Acinetobacter baumannii clinical isolates in Spain. Clinical Microbiology and Infection.

2007;13(12):1192-8.

[29] Wang S, Sheng W-H, Chang Y-Y, Wang L, Lin H, Chen M, et al.

Healthcare-associated outbreak due to pan-drug resistant Acineto- bacter baumannii in a surgical intensive care unit. Journal of Hospi- tal Infection. 2003;53(2):97-102.

[30] Hujer KM, Hujer AM, Hulten EA, Bajaksouzian S, Adams JM, Donskey CJ, et al. Analysis of antibiotic resistance genes in multidrug-resistant Acinetobacter sp. isolates from military and ci- vilian patients treated at the Walter Reed Army Medical Center.

Antimicrobial agents and chemotherapy. 2006;50(12):4114-23.

[31] Simhon A, Rahav G, Shazberg G, Block C, Bercovier H, Shapiro M. Acinetobacter baumannii at a tertiary-care teaching hospital in Jerusalem, Israel. Journal of clinical microbiology. 2001;39(1):389- 91.

[32] SA J-M, Sh B. Risk factors and antimicrobial susceptibility in ventilator associated pneumonia: a brief report. Tehran University Medical Journal TUMS Publications. 2012;70(9):577-82.

(8)

[33] Zarrilli R, Giannouli M, Tomasone F, Triassi M, Tsakris A. Car- bapenem resistance in Acinetobacter baumannii: the molecular epi- demic features of an emerging problem in health care facilities. The Journal of Infection in Developing Countries. 2009;3(05):335-41.

[34] Szejbach A, Mikucka A, Bogiel T, Gospodarek E. Usefulness of phenotypic and genotypic methods for metallo-beta-lactamases detection in carbapenem-resistant Acinetobacter baumannii strains.

Medical science monitor basic research. 2013;19:32.

[35] Sengstock D, Thyagarajan R, Apalara J, Mira A, Chopra T, Kaye K. Multidrug-resistant Acinetobacter baumannii: an emerging pathogen among older adults in community hospitals and nursing homes. Clinical Infectious Diseases. 2010;50(12):1611-6.

[36] Josheghani SB, Moniri R, Firoozeh F, Sehat M, Dastehgoli K, Koosha H, et al. Emergence of bla OXA-Carrying Carbapenem Re- sistance in Multidrug-Resistant Acinetobacter baumannii in the In- tensive Care Unit. Iranian Red Crescent Medical Journal.

2017;19(5).

[37] Azimi L, Lari AR, Talebi M, Namvar AE, Jabbari M. Comparison between phenotypic and PCR for detection of OXA-23 type and metallo-beta-lactamases producer Acinetobacter spp. GMS hygiene and infection control. 2013;8(2).

[38] Shoja S, Moosavian M, Rostami S, Abbasi F, Tabatabaiefar MA, Peymani A. Characterization of oxacillinase and metallo-β- lactamas genes and molecular typing of clinical isolates of Acine- tobacter baumannii in Ahvaz, South-West of Iran. Jundishapur journal of microbiology. 2016;9(5).

[39] Shahcheraghi F, Abbasalipour M, Feizabadi M, Ebrahimipour G, Akbari N. Isolation and genetic characterization of metallo-β- lactamase and carbapenamase producing strains of Acinetobacter baumannii from patients at Tehran hospitals. Iranian journal of microbiology. 2011;3(2):68.

[40] NajarPeerayeh S, Karmostaji A, Salmanian AH. Distribution of OXA-type class D β-lactamase genes among nosocomial multi

drug resistant Acinetobacter baumannii isolated in Tehran hospitals. Jundishapur Journal of Microbiology. 2013;6(5):1-5.

[41] Salimizand H, Modarresi F, Azizi O, Mansouri S, Rahmati M, Barkhordari K. Antimicrobial Profile and Phenotypic Metallo-β- Lactamase Detection of Acinetobacter baumannii Isolated From Clinical and Environmental Specimens. Zahedan Journal of Re- search in Medical Sciences. 2015;17(6).

[42] Karmostaji A, Javadpour S, Davoodian P, Moradi N. In vitro activity of tigecycline and colistin against clinical isolates of Acineto- bacter baumannii in hospitals in Tehran and Bandar-Abbas, Iran.

Electronic physician. 2014;6(3):919.

[43] Khorsi K, Messai Y, Hamidi M, Ammari H, Bakour R. High prevalence of multidrug-resistance in Acinetobacter baumannii and dis- semination of carbapenemase-encoding genes blaOXA-23-like, blaOXA-24-like and blaNDM-1 in Algiers hospitals. Asian Pacific journal of tropical medicine. 2015;8(6):438-46.

[44] Srinivasan VB, Rajamohan G, Pancholi P, Stevenson K, Tadesse D, Patchanee P, et al. Genetic relatedness and molecular characterization of multidrug resistant Acinetobacter baumannii isolated in central Ohio, USA. Annals of clinical microbiology and antimicro- bials. 2009;8(1):21.

[45] Lin M-F, Chang K-C, Lan C-Y, Chou J, Kuo J-W, Chang C-K, et al. Molecular epidemiology and antimicrobial resistance determinants of multidrug-resistant Acinetobacter baumannii in five proximal hospitals in Taiwan. Japanese journal of infectious diseases. 2011;64(3):222-7.

[46] Zarrilli R, Crispino M, Bagattini M, Barretta E, Di Popolo A, Tri- assi M, et al. Molecular epidemiology of sequential outbreaks of Acinetobacter baumannii in an intensive care unit shows the emergence of carbapenem resistance. Journal of clinical microbiology.

2004;42(3):946-53.

DISCLAIMER: The above article has been published in Epub (ahead of print) on the basis of the materials provided by the author. The Editorial Department reserves the right to make minor modifications for further improvement of the manuscript.

View publication stats View publication stats