Antibiotic resistance assessment, and genotypic and phenotypic detection of norA efflux pump in methicillin and ciprofloxacin resistant Staphylococcus aureus isolates
Subject Areas : Molecular Microbiology
Samaneh sadat Kazemi
1
(
MS.c., Department of Biotechnology, Faculty of Advanced Science & Technology, Pharmaceutical Sciences branch, Islamic Azad University, Tehran, Iran.
)
Fahimeh Nemati Mansour
2
(
Assistant Professor, Department of Biotechnology, Faculty of Advanced Science & Technology, Pharmaceutical Sciences branch, Islamic Azad University, Tehran, Iran.
)
Amir Mirzaei
3
(
Assistant Professor., Young Researchers and Elite Club, East Tehran branch, Islamic Azad University, Tehran, Iran.
)
Fatemeh Ashrafi
4
(
Assistant Professor, Department of Biology, Tehran North branch, Islamic Azad University, Tehran, Iran.
)
Keywords: Staphylococcus aureus, Efflux pump, norA, Ciprofloxacin resistance,
Abstract :
Background & Objectives: Staphylococcus aureus is one of the major causes of nosocomial infections throughout the world. Efflux pumps such as norA play a major role in the development of resistance to different antibiotics in this bacteria. The aim of this study was evaluation of the antibiotic resistance and detection of efflux pump (norA) in methicillin resistant (MRSA) and ciprofloxacin resistant S. aureus isolates using genotypic and phenotypic methods. Materials & Methods: During this sectional study, 250 clinical samples were collected from different hospitals in Tehran, Iran. S. aureus isolates were identified and the antimicrobial susceptibility patterns were determined. Ciprofloxacin minimum inhibitory concentration (MIC) was determined in both MRSA and ciprofloxacin resistant isolates. Furthermore, the presence of norA efflux pump gene in MRSA and ciprofloxacin resistant isolates was assessed phenotypically using ciprofloxacin and ethidium bromide MIC, with CCCP as efflux pump inhibitor, and genetically using PCR method. Results: Totally, 50 S. aureus isolates were recovered. The results of antibiotic susceptibility tests showed that 34 isolates (68%) were resistant to methicillin, of which 12 isolates (24%) were resistant to ciprofloxacin, as well. Moreover, all the MRSA- ciprofloxacin resistant strains harbored the norA gene and active efflux pump. Conclusion: The results showed the correlation between ciprofloxacin resistance and norA efflux pump gene in MRSA isolates. Development of efflux pump inhibitors can be useful in the control of MRSA ciprofloxacin resistant strains.
1. Iliyasu G, Daiyab FM, Tiamiyu AB, Abubakar S, Habib ZG, Sarki AM, Habib AG. Nosocomial infections and resistance pattern of common bacterial isolates in an intensive care unit of a tertiary hospital in Nigeria: A 4-year review. J Crit Care. 2016; 34: 116-120.
2. Changchien CH, Chen SW, Chen YY, Chu C. Antibiotic susceptibility and genomic variations in Staphylococcus aureus associated with Skin and Soft Tissue Infection (SSTI) disease groups. BMC Infect Dis. 2016; 10:16(1): 276.
3. Davoodabadi F, Mobasherizadeh S, Mostafavizadeh K, Shojaei H, Havaei SA, Koushki AM, Moghadasizadeh Z, Meidani M, Shirani K. Nasal colonization in children with communityacquired methicillin-resistant Staphylococcus aureus. Adv Biomed Res. 2016; 11; 5: 86.
4. Mustapha M, Bukar-Kolo YM, Geidam YA, Gulani IA. Phenotypic and genotypic detection of methicillin-resistant Staphylococcus aureus in hunting dogs in Maiduguri metropolitan, Borno State, Nigeria. Vet World. 2016; 9(5): 501-506.
5. Garonzik SM, Lenhard JR, Forrest A, Holden PN, Bulitta JB, Tsuji BT. Defining the active fraction of daptomycin against Methicillin-Resistant Staphylococcus aureus (MRSA) using a pharmacokinetic and pharmacodynamic approach. PLoS One. 2016; 11(6): e0156131.
6. Borgmann S, Rieß B, von Wernitz-Keibel T, Bühler M, Layer F, Strommenger B. Recovery of a 10-year-old girl from methicillin-resistant Staphylococcus aureus sepsis in response to low-dose ceftaroline treatment. Ther Clin Risk Manag. 2016; 11; 12: 749-753.
7. K. Poole. Efflux pumps as antimicrobial resistance mechanisms. Ann Med. 2007; 39: 162-176.
8. Li XZ, Nikaido H. Efflux-mediated drug resistance in bacteria. Drugs. 2004; 64; 159-204.
9. Kosmidis C, Schindler BD, Jacinto PL, Patel D, Bains K, Seo SM, Kaatz GW. Expression of multidrug resistance efflux pump genes in clinical and environmental isolates of Staphylococcus aureus. Int J Antimicrob Agents. 2012; 40: 204-209.
10. Paulsen IT, Lewis K. Microbial multidrug efflux. Horizon Scientific. 2002; 3(2): 143-144.
11. Li XZ, Nikaido H. Efflux-mediated drug resistance in bacteria: an update. Drugs. 2009; 69: 1555-1623.
12. Ding Y, Onodera Y, Lee JC, Hooper DC. NorB, an efflux pump in Staphylococcus aureus strain MW2, contributes to bacterial fitness in abscesses. J Bacteriol. 2008; 190: 7123-7129.
13. Kumar A, Schweizer HP. Bacterial resistance to antibiotics: active efflux and reduced uptake. Adv Drug Deliv Rev. 2005; 57: 1486-1513.
14. Motallebi M, Jabalameli F, Asadollahi K, Taherikalani M, Emaneini M. Spreading of genes encoding enterotoxins, haemolysins, adhesin and biofilm among methicillin resistant Staphylococcus aureus strains with staphylococcal cassette chromosome mec type IIIA isolated from burn patients. Microb Pathog. 2016; 97: 34-37.
15. Ruu HD, Cornelis V, Christel D, Michele B, Nancy L, Etienne EE, Stobberingh A. Rapid detection of panton-valentine leukocidin from clinical isolates of Staphylococcus aureus strains by Real-Time PCR. J FEMS Microbial Letters. 2004; 240(2): 225-228.
16. Clinical and laboratory standards institute (CLSI), 2006. Performance standards for antimicrobial susceptibility testing; 16th informational supplement. CLSI, Wayne, Pa. M100-S16, 26, no. 3. 2006.
17. Wielders CL1,Fluit AC ,Brisse S ,Verhoef J ,Schmitz FJ .mecA gene is widely disseminated in Staphylococcus aureus population. J Clin Microbiol. 2002; 40(11): 3970-3975.
18. Ding Y, Onodera Y, Lee JC. NorB, an efflux pump in Staphylococcus aureus strain MW2, contributes to bacterial fitness in abscesses. J Bacteriol. 2008; 190(21): 7123-7129.
19. Ardebili A, Talebi M, Azimi L, Rastegar Lari A. Effect of efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone on the minimum inhibitory concentration of ciprofloxacin in Acinetobacter baumannii clinical isolates. Jundishapur J Microbiol. 2014; 7(1): e8691.
20. Ghosh S, Banerjee M. Methicillin resistance & amp; inducible clindamycin resistance in Staphylococcus aureus. Indian J Med Res. 2016; 143(3): 362-364.
21. Pourmand MR, Yousefi M ,Salami SA ,Amini M .Evaluation of expression of NorA efflux pump in ciprofloxacin resistant Staphylococcus aureus against hexa hydroquinoline derivative by Real-time PCR. Acta Med Iran. 2014; 52(6): 424-429.
22. Ardebili A, Lari AR, Beheshti M, Lari ER. Association between mutations in gyrA and parC genes of Acinetobacter baumannii clinical isolates and ciprofloxacin resistance. Iran J Basic Med Sci. 2015; 18(6): 623-626.
23. Piddock LJV. Clinically relevant chromosomally encoded multidrug resistance efflux pumps in bacteria. Clin Microbiol Rev. 2006; 19: 382-402.
24. Saiful AJ, Mastura M, Zarizal S, Mazurah MI, Shuhaimi M, Ali AM. Efflux genes and active efflux activity detection in Malaysian clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). J Basic Microbiol. 2008; 48(4): 245-251.
25. Patel D, Kosmidis C ,Seo SM ,Kaatz GW .Ethidium bromide MIC screening for enhanced efflux pump gene expression or efflux activity in Staphylococcus aureus. Antimicrob Agents Chemother. 2010; 54(12): 5070-5073.
26. Costa SS, Junqueira E ,Palma C ,Viveiros M ,Melo-Cristino J ,Amaral L ,Couto I. Resistance to antimicrobials mediated by efflux pumps in Staphylococcus aureus. Antibiotics (Basel). 2013; 2(1): 83-99.
27. Couto I, Costa SS ,Viveiros M ,Martins M ,Amaral L .Efflux-mediated response of Staphylococcus aureus exposed to ethidium bromide. J Antimicrob Chemother. 2008; 62(3): 504-513.
1. Iliyasu G, Daiyab FM, Tiamiyu AB, Abubakar S, Habib ZG, Sarki AM, Habib AG. Nosocomial infections and resistance pattern of common bacterial isolates in an intensive care unit of a tertiary hospital in Nigeria: A 4-year review. J Crit Care. 2016; 34: 116-120.
2. Changchien CH, Chen SW, Chen YY, Chu C. Antibiotic susceptibility and genomic variations in Staphylococcus aureus associated with Skin and Soft Tissue Infection (SSTI) disease groups. BMC Infect Dis. 2016; 10:16(1): 276.
3. Davoodabadi F, Mobasherizadeh S, Mostafavizadeh K, Shojaei H, Havaei SA, Koushki AM, Moghadasizadeh Z, Meidani M, Shirani K. Nasal colonization in children with communityacquired methicillin-resistant Staphylococcus aureus. Adv Biomed Res. 2016; 11; 5: 86.
4. Mustapha M, Bukar-Kolo YM, Geidam YA, Gulani IA. Phenotypic and genotypic detection of methicillin-resistant Staphylococcus aureus in hunting dogs in Maiduguri metropolitan, Borno State, Nigeria. Vet World. 2016; 9(5): 501-506.
5. Garonzik SM, Lenhard JR, Forrest A, Holden PN, Bulitta JB, Tsuji BT. Defining the active fraction of daptomycin against Methicillin-Resistant Staphylococcus aureus (MRSA) using a pharmacokinetic and pharmacodynamic approach. PLoS One. 2016; 11(6): e0156131.
6. Borgmann S, Rieß B, von Wernitz-Keibel T, Bühler M, Layer F, Strommenger B. Recovery of a 10-year-old girl from methicillin-resistant Staphylococcus aureus sepsis in response to low-dose ceftaroline treatment. Ther Clin Risk Manag. 2016; 11; 12: 749-753.
7. K. Poole. Efflux pumps as antimicrobial resistance mechanisms. Ann Med. 2007; 39: 162-176.
8. Li XZ, Nikaido H. Efflux-mediated drug resistance in bacteria. Drugs. 2004; 64; 159-204.
9. Kosmidis C, Schindler BD, Jacinto PL, Patel D, Bains K, Seo SM, Kaatz GW. Expression of multidrug resistance efflux pump genes in clinical and environmental isolates of Staphylococcus aureus. Int J Antimicrob Agents. 2012; 40: 204-209.
10. Paulsen IT, Lewis K. Microbial multidrug efflux. Horizon Scientific. 2002; 3(2): 143-144.
11. Li XZ, Nikaido H. Efflux-mediated drug resistance in bacteria: an update. Drugs. 2009; 69: 1555-1623.
12. Ding Y, Onodera Y, Lee JC, Hooper DC. NorB, an efflux pump in Staphylococcus aureus strain MW2, contributes to bacterial fitness in abscesses. J Bacteriol. 2008; 190: 7123-7129.
13. Kumar A, Schweizer HP. Bacterial resistance to antibiotics: active efflux and reduced uptake. Adv Drug Deliv Rev. 2005; 57: 1486-1513.
14. Motallebi M, Jabalameli F, Asadollahi K, Taherikalani M, Emaneini M. Spreading of genes encoding enterotoxins, haemolysins, adhesin and biofilm among methicillin resistant Staphylococcus aureus strains with staphylococcal cassette chromosome mec type IIIA isolated from burn patients. Microb Pathog. 2016; 97: 34-37.
15. Ruu HD, Cornelis V, Christel D, Michele B, Nancy L, Etienne EE, Stobberingh A. Rapid detection of panton-valentine leukocidin from clinical isolates of Staphylococcus aureus strains by Real-Time PCR. J FEMS Microbial Letters. 2004; 240(2): 225-228.
16. Clinical and laboratory standards institute (CLSI), 2006. Performance standards for antimicrobial susceptibility testing; 16th informational supplement. CLSI, Wayne, Pa. M100-S16, 26, no. 3. 2006.
17. Wielders CL1,Fluit AC ,Brisse S ,Verhoef J ,Schmitz FJ .mecA gene is widely disseminated in Staphylococcus aureus population. J Clin Microbiol. 2002; 40(11): 3970-3975.
18. Ding Y, Onodera Y, Lee JC. NorB, an efflux pump in Staphylococcus aureus strain MW2, contributes to bacterial fitness in abscesses. J Bacteriol. 2008; 190(21): 7123-7129.
19. Ardebili A, Talebi M, Azimi L, Rastegar Lari A. Effect of efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone on the minimum inhibitory concentration of ciprofloxacin in Acinetobacter baumannii clinical isolates. Jundishapur J Microbiol. 2014; 7(1): e8691.
20. Ghosh S, Banerjee M. Methicillin resistance & amp; inducible clindamycin resistance in Staphylococcus aureus. Indian J Med Res. 2016; 143(3): 362-364.
21. Pourmand MR, Yousefi M ,Salami SA ,Amini M .Evaluation of expression of NorA efflux pump in ciprofloxacin resistant Staphylococcus aureus against hexa hydroquinoline derivative by Real-time PCR. Acta Med Iran. 2014; 52(6): 424-429.
22. Ardebili A, Lari AR, Beheshti M, Lari ER. Association between mutations in gyrA and parC genes of Acinetobacter baumannii clinical isolates and ciprofloxacin resistance. Iran J Basic Med Sci. 2015; 18(6): 623-626.
23. Piddock LJV. Clinically relevant chromosomally encoded multidrug resistance efflux pumps in bacteria. Clin Microbiol Rev. 2006; 19: 382-402.
24. Saiful AJ, Mastura M, Zarizal S, Mazurah MI, Shuhaimi M, Ali AM. Efflux genes and active efflux activity detection in Malaysian clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). J Basic Microbiol. 2008; 48(4): 245-251.
25. Patel D, Kosmidis C ,Seo SM ,Kaatz GW .Ethidium bromide MIC screening for enhanced efflux pump gene expression or efflux activity in Staphylococcus aureus. Antimicrob Agents Chemother. 2010; 54(12): 5070-5073.
26. Costa SS, Junqueira E ,Palma C ,Viveiros M ,Melo-Cristino J ,Amaral L ,Couto I. Resistance to antimicrobials mediated by efflux pumps in Staphylococcus aureus. Antibiotics (Basel). 2013; 2(1): 83-99.
27. Couto I, Costa SS ,Viveiros M ,Martins M ,Amaral L .Efflux-mediated response of Staphylococcus aureus exposed to ethidium bromide. J Antimicrob Chemother. 2008; 62(3): 504-513.
