مقاومت آنتی‌بیوتیکی وابسته به پمپ افلاکس در جدایه های بالینی و محیطی اسینتوباکتر بامانی
محورهای موضوعی : میکروب شناسی پزشکی
مرجان شاه ایلی
1
(دانشجوی دکتری، دانشگاه آزاد اسلامی، دانشگاه علوم و تحقیقات فارس، گروه میکروب شناسی)
مجید باصری صالحی
2
(دانشیار، دانشگاه آزاد اسلامی، واحد کازرون، گروه میکروب شناسی)
نیما بهادر
3
(دانشیار، دانشگاه آزاد اسلامی، واحد شیراز، گروه میکروب شناسی)
کلید واژه: اسینتوباکتر بامانی, پمپ افلاکس, ژن adeABC, PAβN,
چکیده مقاله :
سابقه و هدف: اعضای جنس اسینتوباکتر فلور طبیعی و عامل عفونت‌های فرصت طلب بیمارستانی هستند. هدف از این مطالعه بررسی کسب مقاومت دارویی از طریق پمپ افلاکس در سویه های اسینتوباکتر بامانی مقاوم به چند دارو و نقش فنیل آرژنین بتا نفتیل آمید (PAβN) به عنوان مهار کننده پمپ افلاکس بود.مواد و روش‌ها: در این مطالعه مقطعی-توصیفی 83 جدایه بالینی و 62 جدایه محیطی جمع آوری گردید. شناسایی باکتری‌های جدا شده با آزمون های استاندارد بیوشیمیایی و پرایمر اختصاصی ژن های 16S rRNA و 16S-23S rRNA با روش واکنش زنجیره‌ای پلی‌مراز (PCR) انجام شد. آزمون حساسیت آنتی‌بیوتیکی به روش دیسک دیفیوژن وسنجش MIC با توجه به استاندارد CLSI انجام گردید. ارزیابی فعالیت پمپ افلاکس با استفاده از مهار کننده PAβN و فراوانی ژن adeABC با استفاده از روش PCR مورد بررسی قرارگرفت.یافته‌ها: از مجموع نمونه های مورد بررسی، در 56 (67.47%) نمونه بالینی و 34 (54.84%) نمونه محیطی باکتری اسینتوباکتر بامانی تشخیص داده شد. بر اساس نتایج آنتی‌بیوگرام 100% نمونه‌ها به سفوتاکسیم و سفتری‌آکسون مقاومت نشان دادند. تنها 7.13% از نمونه‌های بالینی و 5.89% از نمونه‌های محیطی مقاومت چندگانه آنتی بیوتیکی را نداشتند. شیوع adeABC در نمونه های بالینی به ترتیب 69.60 %، 82.14%، 76.80 % و در نمونه های محیطی به ترتیب 50 %، 85.30% و 73.50 % بوده است. همچنین حداقل غلظت بازدارندگی تتراسیکلین در حضور PAβN در تمام جدایه‌های مقاوم با کاهش همراه بود.نتیجه گیری: نتایج نشان داد که پمپ افلاکس می‌تواند در مقاومت آنتی‌بیوتیکی جدایه‌های اسینتوباکتر بامانی نقش داشته باشد. همچنین به دلیل مقاومت جدایه‌های اسینتوباکتر بامانی به سفالوسپورین‌ها، پیشنهاد می‌شود از تجویز آن‌ها خودداری گردد.
Background & Objectives: Members of Acinetobacter genus are normal flora and the causal agent of the opportunistic nosocomial infections. The aim of this study was to investigate the ability to acquire drug resistance through efflux pump mechanism in multidrug-resistant Acinetobacter spp. and the role of PAβN (Phenyl Arginin β naphtyamid) as an efflux pump inhibitor. Materials & Methods: In the cross-sectional descriptive study 83 clinical isolates and 62 environmental isolates were collected. Identification of the isolated bacteria was carried out by standard biochemical experiments and polymerase chain reaction (PCR) using specific primers of 16S rRNA and 16S-23S rRNA. Antibiotic susceptibility tests were conducted through disc diffusion and MIC test based on CLSI guidelines. The activity of the efflux pump was evaluated using the PaβN, and the adeABC gene frequency was assessed by PCR. Results: Of the total of the samples evaluated, 56 (67.47%) clinical isolates and 34 (54.84%) environmental isolates were identified as Acinetobacter spp. In accordance with the results of the antibiogram test, 100% of the isolates were resistant to cefotaxime and ceftriaxone. Only 7.13% of the clinical isolates and 5.89% of the environmental isolates were not multidrug-resistant (MDR). Prevalence of the adeB was 69.60%, 82.14%, and 76.80% in the clinical isolates and 50%, 85.30%, and 73.50% in the environmental isolates, respectively. Furthermore, tetracycline MIC was decreased in all resistant isolates in the presence of PAβN. Conclusion: The results showed that the efflux pump could play a role in antibiotic resistance in Acinetobacter spp. isolates. Owing to the resistance of Acinetobacter isolates against cephalosporine, it is suggested to avoid prescribing.
pumps in Acinetobacter spp. isolated from inpatients. J Mazandaran Univ Med Sci. 2015; 25(126):
1-10. [In Persian]
2. Nikasa P, Abdiaali E, Bahar MA, Niknam V. Antibiotic resistance of clinical Acinetobacter
baumannii species in the presence of efflux pump inhibitor. J Infec Dise Trop Med. 2008; 13(42):
19-23. [In Persian]
3. Valadkhani F, Zeighami H. Efflux pump systems in bacteria. Lab Diagn. 2016; 8(33): 35-43.
[In Persian]
4. Godarzi H, Doraghi M, Dabiri H, Ghaladvand Z. Examination of expression and function of efflux
pumps gene of non-clonal multidrug-resistant Acinetobacter baumannii isolates. J Res Med Sc.
2013; 37(2): 107-112. [In Persian]
5. AfsharYavari S, Rota S, Caglar K, Fidan I. Efflux pump inhibitors effect on the minimum
inhibitory concentration of Benzalkonium Chloride and Chlorhexidine in Acinetobacter baumannii
strains isolated from hospitalized inpatients in Gazi hospital, Ankara. J Urmia Univ Med. 2016; 27
(5): 393-401. [In Persian]
6. Akpangodwin U, Illyasu M. Effect of length of soil sample storage on soil microbiological and
bochemical properties in Uyo, Nigeria. J Global Agric Rec. 2015; 3(3): 25-37.
7. Xin F, Cai D, Sun Y, Guo D, Wu Z, Jiang D. Exploring the diversity of Acinetobacter populations
in river water with genus-specific primers and probes. J Gen Appl Microbiol. 2014; 60(2): 51-58.
8. Mahon CR, Lehman OC, Manuselis G. Textbook of diagnostic microbiology fifth edition.
Saunders. 2015; 494-512.
9. Krsek M, Wellington EMH. Comparison of different methods for the isolation and purification of
total community DNA from soil. J Microbiol Methods. 1999; 39(1): 1-16.
10. Chen TL, Siu LK, Wu RCC, Shaio MF, Huang LY, Fung CP, Lee CM, Cho WL. Comparison of
one-tube multiplex PCR, automated ribotyping and intergenic spacer (ITS) sequencing for rapid
identification of Acinetobacter baumannii. Clin Microbiol Infect. 2007; 13(8): 801-806.
11. Patel JB, Patel R, Weinstein MP. Performance standards for antimicrobial susceptibility testing.
M100-S23. 2017; p: 66-67.
12. Japoni-Nejad AR, Sofian M, Ghaznavi-Rad E. Molecular detection of AdeABC efflux pump
genes in clinical isolates of Acinetobacter baumannii and their contribution in imipenem resistance. Iran South Med J. 2014; 17(5): 815-823.
13. Jassim KA, Ghaima KK, Saadedin SMK. AdeABC efflux pump genes in multidrug resistant
Acinetobacter baumannii isolates. Avicenna J Clin Microb Infec. 2016; 3(4):1-6.
14. Angoti G, Bandehpour M, Goudarzi H, Zarringhalammoghaddam M, Kouchaki A. Detection of
efflux pump genes (adeA, adeB, adeC and adeM) in Acinetobacter baumannii isolated from
hospitalize patients, North-west of Iran. Infect Epidemiol Med. 2016; 2(4): 8-11.
15. AfsharYavari S, Rota S, Caglar K, Fidan I. Determination of resistance pattern of isolated
Acinetobacter baumannii from intensive care unite (ICUS) in Gazi hospital, Ankara. J Urmia
Nursing Midwifery. 2016; 13(10): 912-918. [In Persian]
16. Dijkshoorn L, Nemec A, Seifert H.An increasing threat in hospitals: multidrug resistant
Acinetobacter baumannii. Nat Rev Microbiol. 2007; 5(12): 939-951.
17. Jafari S, Najafipour S, Kargar M, Abdollahy A, Mardaneh J, Fasihy Ramandy M,
Abdollahi-Kheirabadi S, Moraveje A. Phenotypical evaluation of multi-drug resistant Acinetobacter
baumannii. J Fasa Uni Med Sci. 2012; 2l(4): 254-258.
18. Vafaei S, Mirnejad R, Amirmozaffari N, Imanifoladi AA, Masjedian F. Antibiotic resistance
pattern and extended β lactamase frequency in Acinetobacter baumannii strains isolates from
clinical samples by phenotypic method. J Infec Dise Trop Med. 2013; 18(61): 39-44. [In Persian]
19. Mirnejad R, Mostofi S, Masjedian F. Evaluation of role of class 2 integron in the antibiotic
susceptibility pattern of Acinetobacter baumannii strains isolated from Tehran hospital. Sci J
Hamdan Univ Med Sci. 2011; 18(4): 22-28. [In Persian]
20. Golestani F, javadpour S, Kafilzadeh F, Ghalandarzadeh Daryaei Z. Determination of antibiotic
resistance pattern and frequency of blaVIM & blaIMP genes in clinical isolates of Acinetobacter species
in Bandar Abbass. J Microb World. 2018; 10(4): 229-309. [In Persian]
21. Mirnejad R, Mostofi S, Masjedian F. Antibiotic resistance and carriage class 1 and 2 integrons in
clinical isolates of Acinetobacter baumannii from Tehran, Iran. Asian Pac J Trop Biomed. 2013; 3
(2): 140-145.
22. Su-ying Z, Dong-yang J, Peng-cheng X, Yi-kai Z, Heng-fang S, Hui-ling C, Qian W. An
investigation of drug-resistant Acinetobacter baumannii infections in a comprehensive hospital of
East China. Ann Clin Microbiol Antimicrob. 2015; 14: 7.
23. Taherikalani M, Maleki A, Sadeghifard N, Mohammadzadeh D, Soroush S, Asadollahi P,
Asadollahi K, Emaneini M. Dissemination of class 1, 2 and 3 integrons among different multidrug
resistant isolates of Acinetobacter baumanniiin Tehran hospitals, Iran. Pol J Microbiol. 2011; 60(2):
169-174.
24. Ghajavand H, Havaei A, Nasr Esfahani B, Fazeli H, Moghim S. Frequency of MDR Acinetobacter
baumannii isolates in Intensive Care Units (ICU) of Isfahan hospitals by molecular method and their
antimicrobial resistance pattern. J Isfahan Med Sch. 2014; 32(295): 1-11. [In Persian]
25. Gholami M, Hashemi A, Hakemi-Vala M, Goudarzi H, Hallajzadeh M. Efflux pump inhibitor
phenylalanine-arginine Β-Naphthylamide effect on the minimum inhibitory concentration of
imipenemin Acinetobacter baumannii strains isolated from hospitalized patients in Shahid Motahari
Burn Hospital, Tehran, Iran. Jundishapur J Microbiol. 2015; 8(10): 1-7.
26. Khayat H, Sadeghifard N, Pakzad I, Azimi L, Delfani S, Sayehmiri K, Soroush S, Bogdanovic L,
Taherikalani M. Determination of different fluoroquinolone mechanisms among clinical isolates of
Acinetobacter baumannii in Tehran, Iran . Iran Red Crescent Med J. 2017; 19(9): 1-8.
27. Lin MF, Lin YY, Tu CC, Lan CY. Distribution of different efflux pump genes in clinical isolates
of multidrug-resistant Acinetobacter baumannii and their correlation with antimicrobial resistance.
J Microbiol Immunol Infect. 2017; 50(2): 224-231.
28. Rizzo C, Marascio N, Zicca E, Pavia G, Quirino A, Lamberti AG, Liberto MC, Focà A. Efflux
pump AdeABC assessment in Acinetobacter baumannii strains isolated in a teaching hospital. J
Med Microb Diag. 2016; 5(3): 1-3.
29. Sarhaddi N, Soleimanpour S, Farsiani H, Mosavatd A, Dolatabadi S, Salimizand H, Amel
Jamehdar S. Elevated prevalence of multidrug-resistant Acinetobacter baumannii with extensive
genetic diversity in the largest burn center of northeast Iran. J Glob Anti Microb Re. 2017; 8: 60-66.
30. Pannek S, Higgins PG, Steinke P, Jonas D, Akova M, Bohnert JA, Seifert H, Kern WV. Multidrug
efflux inhibition in Acinetobacter baumannii: comparison between 1-(1-naphthylmethyl)-piperazine
and phenyl-arginine-b-naphthylamide. J Antimicrob Chemother. 2006; 57(5): 970-974.
31. Qingya D, Mingxiang Z, Jun L, Haichen W, Yongmei H, Wen’en L. AdeABC efflux pump and
resistance of Acinetobacter baumannii against carbapenem. J Cent South Univ. 2017; 42(4):
426-433.
32. Peleg AY, Adams J, Paterson DL. Tigecycline efflux as a mechanism for non-susceptibility in
Acinetobacter baumannii. Antimicrob Agents Chemother. 2007; 51(6): 2065-2069
_||_
pumps in Acinetobacter spp. isolated from inpatients. J Mazandaran Univ Med Sci. 2015; 25(126):
1-10. [In Persian]
2. Nikasa P, Abdiaali E, Bahar MA, Niknam V. Antibiotic resistance of clinical Acinetobacter
baumannii species in the presence of efflux pump inhibitor. J Infec Dise Trop Med. 2008; 13(42):
19-23. [In Persian]
3. Valadkhani F, Zeighami H. Efflux pump systems in bacteria. Lab Diagn. 2016; 8(33): 35-43.
[In Persian]
4. Godarzi H, Doraghi M, Dabiri H, Ghaladvand Z. Examination of expression and function of efflux
pumps gene of non-clonal multidrug-resistant Acinetobacter baumannii isolates. J Res Med Sc.
2013; 37(2): 107-112. [In Persian]
5. AfsharYavari S, Rota S, Caglar K, Fidan I. Efflux pump inhibitors effect on the minimum
inhibitory concentration of Benzalkonium Chloride and Chlorhexidine in Acinetobacter baumannii
strains isolated from hospitalized inpatients in Gazi hospital, Ankara. J Urmia Univ Med. 2016; 27
(5): 393-401. [In Persian]
6. Akpangodwin U, Illyasu M. Effect of length of soil sample storage on soil microbiological and
bochemical properties in Uyo, Nigeria. J Global Agric Rec. 2015; 3(3): 25-37.
7. Xin F, Cai D, Sun Y, Guo D, Wu Z, Jiang D. Exploring the diversity of Acinetobacter populations
in river water with genus-specific primers and probes. J Gen Appl Microbiol. 2014; 60(2): 51-58.
8. Mahon CR, Lehman OC, Manuselis G. Textbook of diagnostic microbiology fifth edition.
Saunders. 2015; 494-512.
9. Krsek M, Wellington EMH. Comparison of different methods for the isolation and purification of
total community DNA from soil. J Microbiol Methods. 1999; 39(1): 1-16.
10. Chen TL, Siu LK, Wu RCC, Shaio MF, Huang LY, Fung CP, Lee CM, Cho WL. Comparison of
one-tube multiplex PCR, automated ribotyping and intergenic spacer (ITS) sequencing for rapid
identification of Acinetobacter baumannii. Clin Microbiol Infect. 2007; 13(8): 801-806.
11. Patel JB, Patel R, Weinstein MP. Performance standards for antimicrobial susceptibility testing.
M100-S23. 2017; p: 66-67.
12. Japoni-Nejad AR, Sofian M, Ghaznavi-Rad E. Molecular detection of AdeABC efflux pump
genes in clinical isolates of Acinetobacter baumannii and their contribution in imipenem resistance. Iran South Med J. 2014; 17(5): 815-823.
13. Jassim KA, Ghaima KK, Saadedin SMK. AdeABC efflux pump genes in multidrug resistant
Acinetobacter baumannii isolates. Avicenna J Clin Microb Infec. 2016; 3(4):1-6.
14. Angoti G, Bandehpour M, Goudarzi H, Zarringhalammoghaddam M, Kouchaki A. Detection of
efflux pump genes (adeA, adeB, adeC and adeM) in Acinetobacter baumannii isolated from
hospitalize patients, North-west of Iran. Infect Epidemiol Med. 2016; 2(4): 8-11.
15. AfsharYavari S, Rota S, Caglar K, Fidan I. Determination of resistance pattern of isolated
Acinetobacter baumannii from intensive care unite (ICUS) in Gazi hospital, Ankara. J Urmia
Nursing Midwifery. 2016; 13(10): 912-918. [In Persian]
16. Dijkshoorn L, Nemec A, Seifert H.An increasing threat in hospitals: multidrug resistant
Acinetobacter baumannii. Nat Rev Microbiol. 2007; 5(12): 939-951.
17. Jafari S, Najafipour S, Kargar M, Abdollahy A, Mardaneh J, Fasihy Ramandy M,
Abdollahi-Kheirabadi S, Moraveje A. Phenotypical evaluation of multi-drug resistant Acinetobacter
baumannii. J Fasa Uni Med Sci. 2012; 2l(4): 254-258.
18. Vafaei S, Mirnejad R, Amirmozaffari N, Imanifoladi AA, Masjedian F. Antibiotic resistance
pattern and extended β lactamase frequency in Acinetobacter baumannii strains isolates from
clinical samples by phenotypic method. J Infec Dise Trop Med. 2013; 18(61): 39-44. [In Persian]
19. Mirnejad R, Mostofi S, Masjedian F. Evaluation of role of class 2 integron in the antibiotic
susceptibility pattern of Acinetobacter baumannii strains isolated from Tehran hospital. Sci J
Hamdan Univ Med Sci. 2011; 18(4): 22-28. [In Persian]
20. Golestani F, javadpour S, Kafilzadeh F, Ghalandarzadeh Daryaei Z. Determination of antibiotic
resistance pattern and frequency of blaVIM & blaIMP genes in clinical isolates of Acinetobacter species
in Bandar Abbass. J Microb World. 2018; 10(4): 229-309. [In Persian]
21. Mirnejad R, Mostofi S, Masjedian F. Antibiotic resistance and carriage class 1 and 2 integrons in
clinical isolates of Acinetobacter baumannii from Tehran, Iran. Asian Pac J Trop Biomed. 2013; 3
(2): 140-145.
22. Su-ying Z, Dong-yang J, Peng-cheng X, Yi-kai Z, Heng-fang S, Hui-ling C, Qian W. An
investigation of drug-resistant Acinetobacter baumannii infections in a comprehensive hospital of
East China. Ann Clin Microbiol Antimicrob. 2015; 14: 7.
23. Taherikalani M, Maleki A, Sadeghifard N, Mohammadzadeh D, Soroush S, Asadollahi P,
Asadollahi K, Emaneini M. Dissemination of class 1, 2 and 3 integrons among different multidrug
resistant isolates of Acinetobacter baumanniiin Tehran hospitals, Iran. Pol J Microbiol. 2011; 60(2):
169-174.
24. Ghajavand H, Havaei A, Nasr Esfahani B, Fazeli H, Moghim S. Frequency of MDR Acinetobacter
baumannii isolates in Intensive Care Units (ICU) of Isfahan hospitals by molecular method and their
antimicrobial resistance pattern. J Isfahan Med Sch. 2014; 32(295): 1-11. [In Persian]
25. Gholami M, Hashemi A, Hakemi-Vala M, Goudarzi H, Hallajzadeh M. Efflux pump inhibitor
phenylalanine-arginine Β-Naphthylamide effect on the minimum inhibitory concentration of
imipenemin Acinetobacter baumannii strains isolated from hospitalized patients in Shahid Motahari
Burn Hospital, Tehran, Iran. Jundishapur J Microbiol. 2015; 8(10): 1-7.
26. Khayat H, Sadeghifard N, Pakzad I, Azimi L, Delfani S, Sayehmiri K, Soroush S, Bogdanovic L,
Taherikalani M. Determination of different fluoroquinolone mechanisms among clinical isolates of
Acinetobacter baumannii in Tehran, Iran . Iran Red Crescent Med J. 2017; 19(9): 1-8.
27. Lin MF, Lin YY, Tu CC, Lan CY. Distribution of different efflux pump genes in clinical isolates
of multidrug-resistant Acinetobacter baumannii and their correlation with antimicrobial resistance.
J Microbiol Immunol Infect. 2017; 50(2): 224-231.
28. Rizzo C, Marascio N, Zicca E, Pavia G, Quirino A, Lamberti AG, Liberto MC, Focà A. Efflux
pump AdeABC assessment in Acinetobacter baumannii strains isolated in a teaching hospital. J
Med Microb Diag. 2016; 5(3): 1-3.
29. Sarhaddi N, Soleimanpour S, Farsiani H, Mosavatd A, Dolatabadi S, Salimizand H, Amel
Jamehdar S. Elevated prevalence of multidrug-resistant Acinetobacter baumannii with extensive
genetic diversity in the largest burn center of northeast Iran. J Glob Anti Microb Re. 2017; 8: 60-66.
30. Pannek S, Higgins PG, Steinke P, Jonas D, Akova M, Bohnert JA, Seifert H, Kern WV. Multidrug
efflux inhibition in Acinetobacter baumannii: comparison between 1-(1-naphthylmethyl)-piperazine
and phenyl-arginine-b-naphthylamide. J Antimicrob Chemother. 2006; 57(5): 970-974.
31. Qingya D, Mingxiang Z, Jun L, Haichen W, Yongmei H, Wen’en L. AdeABC efflux pump and
resistance of Acinetobacter baumannii against carbapenem. J Cent South Univ. 2017; 42(4):
426-433.
32. Peleg AY, Adams J, Paterson DL. Tigecycline efflux as a mechanism for non-susceptibility in
Acinetobacter baumannii. Antimicrob Agents Chemother. 2007; 51(6): 2065-2069