ارزیابی فراوانی ژن های مقاومت به متی سیلین (mecA) و پنتون ولنتین لوکوسیدین (pvl) در سویه های استافیلوکوکوس اورئوس جداسازی شده از نمونه های بالینی بیمارستان های شهر رشت
محورهای موضوعی : میکروب شناسی پزشکی
مریم رحیم پور حصاری
1
(
کارشناس ارشد، گروه زیست شناسی، واحد رشت، دانشگاه آزاد اسلامی، رشت
)
امیر میرزایی
2
(
مربی، باشگاه پژوهشگران جوان و نخبگان، واحد تهران شرق، دانشگاه آزاد اسلامی، تهران
)
علی صالح زاده
3
(
استادیار، گروه زیست شناسی، واحد رشت، دانشگاه آزاد اسلامی، رشت
)
کلید واژه: استافیلوکوکوس اورئوس, ژن mecA, ژن pvl,
چکیده مقاله :
سابقه و هدف: استافیلوکوکوس اورئوس یکی از مهم ترین عوامل عفونت بیمارستانی است و امروزه سویه های مقاوم به متی سیلین شیوع زیادی پیدا کرده اند. هم چنین، توکسین پنتون ولنتین لکوسیدین یکی از عوامل مهم بیماری زا در استافیلوکوکوس اورئوس می باشد. این مطالعه با هدف بررسی مقاومت آنتی بیوتیکی، حضور ژن های مقاومت به متی سیلین و پنتون ولنتین لوکوسیدین در میان سویه های بالینی استافیلوکوکوس اورئوس جداسازی شده از بیمارستان های شهر رشت انجام شد. مواد و روش ها: این مطالعه به صورت مقطعی بر روی 250 نمونه بالینی جمع آوری شده از بیمارستان های مختلف شهر رشت در یک سال (1393-1392) انجام شد. در ابتدا جدایه های استافیلوکوکوس اورئوس توسط آزمون های میکروبی شناسایی شدند. حساسیت آنتی بیوتیکی آن ها بر اساس روش استاندارد CLSI تعیین گردید. همچنین، برای ردیابی وجود ژن های مقاومت به متی سیلین (mecA) و پنتون والنتین لوکوسیدین (pvl) از واکنش زنجیره ای پلی مراز استفاده شد. یافته ها: از میان 250 نمونه مورد بررسی، 50 جدایه متعلق به استافیلوکوکوس اورئوس بودند. آزمایش حساسیت میکروبی نشان داد که 34 سویه (68 درصد) از جدایه های استافیلوکوکوس اورئوس مقاوم به متی سیلین بودند. هم چنین فراوانی ژن های mecA و pvl در بین جدایه های استافیلوکوکوس اورئوس به ترتیب 60 درصد (30 نمونه) و 20 درصد (10 نمونه) بود. نتیجه گیری: نتایج این پژوهش افزایش مقاومت استافیلوکوکوس اورئوس نسبت به آنتی بیوتیک های مختلف را نشان داد. این امر می تواند یک هشدار جدی برای درمان عفونت های ناشی از استافیلوکوکوس اورئوس در منطقه مورد بررسی باشد.
Background & Objectives: Staphylococcus aureus is one of the most important nosocomial infection agent and now methicillin-resistant S. aureus have spread. Also, panton-valentin leukocidin is one of important virulence factors in S. aureus. The study was aimed to evaluate the antibiotic resistant, frequency methicillin resistant and panton- valentin leukocidin genes in clinical S. aureus isolates from hospitals of Rasht, Iran. Materials & Methods: This cross-sectional study was carried out on 250 clinical samples collected from hospitals of Rasht during one year (2013-2014). The S. aureus isolates were identified by microbiological methods. Antibiotic sensitivity was performed by CSLI method using disk diffusion method. In addition, the presence of methicillin resistant (mecA) and panton-valentin leukocidin (pvl) genes were evaluated using polymerase chain reaction (PCR). Results: Totally, 50 S. aureus isolates were recovered. The results of antibiotic susceptibility tests showed that 34 out of 50 S. aureus isolates (68%) were resistant to methicillin. In addition, the prevalence of mecA and pvl gene among isolates were 60% (30 isolates) and 20% (10 isolates), respectively. Conclusion: This study showed increased resistance to different antibiotics in S. aureus that is a serious warning to the treatment of infections caused by this bacterium in the region.
1. Malekzade F. Microbiology. 5th ed., University of Tehran press. 2010.
2. Tarai B, Das P, Kumar D. Recurrent challenges for clinicians: emergence of methicillin resistant, vancomycin resistance, and current treatment options. J Lab Physicians. 2013; 5(2): 71-78.
3. Shorr AF. Epidemiology of staphylococcal resistance. Clin Infect Dis. 2007; 45 (Supplement 3): S171-S176.
4. Chambers HF, Deleo FR. Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol. 2009; 7(9): 629-641.
5. Lindsay JA. Evolution of Staphylococcus aureus and MRSA during outbreaks. Infect Genet Evol. 2014; 21: 548-553.
6. Lina G, Piemont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis. 1999; 29(5): 1128-1132.
7. Supersac G, Prevost G, Piemont Y. Sequencing of leucocidin R from Staphylococcus aureus P83 suggests that Staphylococcal leucocidins and gamma-hemolysin are members of a single, two-component family of toxins. Infect Immun. 1993; 61(2): 580-587.
8. Clark J. A brief review of Panton-Valentine leukocidin producing staphylococcal infections in the intensive therapy unit. Curr Anaesthesia and Crit Care. 2002; 19(5): 330-332.
9. Colin DA, Mazurier I, Sire S, Finck-Barbancon V. Interaction of the two components of leukocidin from Staphylococcus aureus with human polymorphonuclear leukocyte membranes: sequential binding and subsequent activation. Infect Immun. 1994; 62(8): 3184-3188.
10. Cupane L, Pugacova N, Berzina D, Cauce V, Gardovska D, Miklasevics E. Patients with Panton-Valentine leukocidin positive Staphylococcus aureus infections run an increased risk of longer hospitalisation. Int J Mol Epidemiol Genet. 2012; 3(1): 48-55.
11. Rossney AS, Shore AC, Morgan PM, Fitzgibbon MM, O'Connell B, Coleman DC. The emergence and importation of diverse genotypes of methicillin-resistant Staphylococcus aureus (MRSA) harboring the Panton-Valentine leukocidin gene (pvl) reveal that pvl is a poor marker for community acquired MRSA strains in Ireland. J Clin Microbiol. 2007; 45(8): 2554-2563.
12. Vandenesch F, Naimi T, Enright MC, Lina G, Nimmo GR, Heffernan H. Community acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leucocidin genes: worldwide emergence. Emerg Infect Dis. 2003; 9(8): 978-984.
13. Brown ML, O'Hara FP, Close NM, Mera RM, Miller LA, Suaya JA. Prevalence and sequence variation of panton-valentine leukocidin in methicillin-resistant and methicillin-susceptible staphylococcus aureus strains in the United States. J Clin Microbiol. 2012; 50(1): 86-90.
14. Santosaningsih D, Santoso S, Budayanti NS, Kuntaman K, Lestari ES, Farida H, Hapsari R, Hadi P, Winarto W, Milheiriço C, Maquelin K, Willemse-Erix D, van Belkum A, Severin JA, Verbrugh HA. Epidemiology of Staphylococcus aureus harboring the mecA or Panton-Valentine leukocidin genes in hospitals in Java and Bali, Indonesia. Am Soc of Trop Med Hyg. 2014; 3(4): 728-734.
15. Hu Q, Cheng H, Yuan W, Zeng F, Shang W, Tang D, Xue W, Fu J, Zhou R, Zhu J, Yang J, Hu Z, Yuan J, Zhang X, Rao Q, Li S, Chen Z, Hu X, Wu X, Rao X. Panton-Valentine Leukocidin (PVL)-positive health care-associated methicillin-resistant Staphylococcus aureus isolates are associated with skin and soft tissue infections and colonized mainly by infective PVL-encoding bacteriophages. J Clin Microbiol. 2015; 53(1): 67-72.
16. Moghadam SO, Havaei SA. Prevalence of methicillin-resistant Staphylococcus aureus carrying panton-valentine leukocidin gene in cutaneous infections in the city of Isfahan. J Med Bacteriol. 2012; 19(2): 9-16.
17. Motamedi H, Rahmat Abadi SS. The Association of panton-valentine leukocidin and mecA genes in methicillin-resistant Staphylococcus aureus isolates from patients referred to educational hospitals in Ahvaz, Iran. Jundishapur J Microbiol. 2015; 8(8): e22021.
18. Lu SY, Chang FY, Cheng CC, Lee KD, Huang YC. Methicillin-resistant Staphylococcus aureus nasal colonization among adult patients visiting emergency department in a medical center in Taiwan. PLoS ONE. 2011; 6(6): e18620.
19. Shakeri F, Golalipour M, Rahimi Alang S, Ghaemi EA. Spa diversity among MRSA and MSSA strains of Staphylococcus aureus in north of Iran. Int J Microbiol. 2010; (2): 351397.
20. Ghaznavi-Rad E, Nor Shamsudin M, Sekawi Z, van Belkum A, Neela V. A simplified multiplex PCR assay for fast and easy discrimination of globally distributed staphylococcal cassette chromosome mec types in meticillin-resistant Staphylococcus aureus. Med Microbiol. 2010; 59(10): 1135-1139.
21. Ohadian Moghadam S, Havaei SA, Pourmand MR. Prevalence of methicillin-resistant Staphylococcus aureus carrying panton– valentine leukocidin gene in cutaneous infections in the city of Isfahan. J Med Bacteriol. 2012; 1(2): 9-16.
22. Zamani A, Sadeghian S, Ghaderkhani J, Alikhani MY, Najafimosleh M, Taghi Goodarzi M. Detection of methicillin-resistance (mec-A) gene in Staphylococcus aureus strains by PCR and determination of antibiotic susceptibility. Ann Microbiol. 2007; 57(2): 273-276.
23. Moradi N, Javadpour S, Karmostaji A. Reduced sensitivity of staphylococcus aureus to vancomycin. Hormozgan Uni Med Sciences. 2011; 15(3): 169-177.
24. Haghgoo S, Moaddab S, Rafi A. Study of antibiotic resistance pattern of Staphylococcus aureus strains isolated from blood cultures in Tabriz Shahid Madani Hospital. J Jundishapur. 2012; 3(2): 383-390.
25. Molla-abbaszadeh H, Mobayen H, Mirzaei H. Identification of pantonvalentine leukocidin (pvl) genes in Staphylococcus aureus isolated from ipatients of Emam Reza and Shohada Hospitals of Tabriz by real-time PCR. Iran J Med Microbiol. 2013; 6(4): 72-80.
26. Khosravi AD, Hoveizavi H, Farshadzadeh Z. The prevalence of genes encoding leukocidins in Staphylococcus aureus strains resistant and sensitive to methicillin isolated from burn patients in Taleghani Hospital, Ahvaz, Iran. Burns. 2012; 38(2): 247-251.
27. Otokunefor K, Sloan T, Kearns AM, James R. Molecular characterization and panton-valentine leucocidin typing of community-acquired methicillin-sensitive Staphylococcus aureus clinical isolates. J Clin Microbiol. 2012; 50(9): 3069-72.
28. Osman NAM, Intisar IE, Mohamed YM, El-Eragi AM, Eldirdery MM, Salih MA. Molecular study of panton-valentine leukocidin genes among Staphylococcus aureus clinical isolates in Khartoum State, Sudan. Am J Microbiol Res. 2015; 3(3): 107-111.
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1. Malekzade F. Microbiology. 5th ed., University of Tehran press. 2010.
2. Tarai B, Das P, Kumar D. Recurrent challenges for clinicians: emergence of methicillin resistant, vancomycin resistance, and current treatment options. J Lab Physicians. 2013; 5(2): 71-78.
3. Shorr AF. Epidemiology of staphylococcal resistance. Clin Infect Dis. 2007; 45 (Supplement 3): S171-S176.
4. Chambers HF, Deleo FR. Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol. 2009; 7(9): 629-641.
5. Lindsay JA. Evolution of Staphylococcus aureus and MRSA during outbreaks. Infect Genet Evol. 2014; 21: 548-553.
6. Lina G, Piemont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis. 1999; 29(5): 1128-1132.
7. Supersac G, Prevost G, Piemont Y. Sequencing of leucocidin R from Staphylococcus aureus P83 suggests that Staphylococcal leucocidins and gamma-hemolysin are members of a single, two-component family of toxins. Infect Immun. 1993; 61(2): 580-587.
8. Clark J. A brief review of Panton-Valentine leukocidin producing staphylococcal infections in the intensive therapy unit. Curr Anaesthesia and Crit Care. 2002; 19(5): 330-332.
9. Colin DA, Mazurier I, Sire S, Finck-Barbancon V. Interaction of the two components of leukocidin from Staphylococcus aureus with human polymorphonuclear leukocyte membranes: sequential binding and subsequent activation. Infect Immun. 1994; 62(8): 3184-3188.
10. Cupane L, Pugacova N, Berzina D, Cauce V, Gardovska D, Miklasevics E. Patients with Panton-Valentine leukocidin positive Staphylococcus aureus infections run an increased risk of longer hospitalisation. Int J Mol Epidemiol Genet. 2012; 3(1): 48-55.
11. Rossney AS, Shore AC, Morgan PM, Fitzgibbon MM, O'Connell B, Coleman DC. The emergence and importation of diverse genotypes of methicillin-resistant Staphylococcus aureus (MRSA) harboring the Panton-Valentine leukocidin gene (pvl) reveal that pvl is a poor marker for community acquired MRSA strains in Ireland. J Clin Microbiol. 2007; 45(8): 2554-2563.
12. Vandenesch F, Naimi T, Enright MC, Lina G, Nimmo GR, Heffernan H. Community acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leucocidin genes: worldwide emergence. Emerg Infect Dis. 2003; 9(8): 978-984.
13. Brown ML, O'Hara FP, Close NM, Mera RM, Miller LA, Suaya JA. Prevalence and sequence variation of panton-valentine leukocidin in methicillin-resistant and methicillin-susceptible staphylococcus aureus strains in the United States. J Clin Microbiol. 2012; 50(1): 86-90.
14. Santosaningsih D, Santoso S, Budayanti NS, Kuntaman K, Lestari ES, Farida H, Hapsari R, Hadi P, Winarto W, Milheiriço C, Maquelin K, Willemse-Erix D, van Belkum A, Severin JA, Verbrugh HA. Epidemiology of Staphylococcus aureus harboring the mecA or Panton-Valentine leukocidin genes in hospitals in Java and Bali, Indonesia. Am Soc of Trop Med Hyg. 2014; 3(4): 728-734.
15. Hu Q, Cheng H, Yuan W, Zeng F, Shang W, Tang D, Xue W, Fu J, Zhou R, Zhu J, Yang J, Hu Z, Yuan J, Zhang X, Rao Q, Li S, Chen Z, Hu X, Wu X, Rao X. Panton-Valentine Leukocidin (PVL)-positive health care-associated methicillin-resistant Staphylococcus aureus isolates are associated with skin and soft tissue infections and colonized mainly by infective PVL-encoding bacteriophages. J Clin Microbiol. 2015; 53(1): 67-72.
16. Moghadam SO, Havaei SA. Prevalence of methicillin-resistant Staphylococcus aureus carrying panton-valentine leukocidin gene in cutaneous infections in the city of Isfahan. J Med Bacteriol. 2012; 19(2): 9-16.
17. Motamedi H, Rahmat Abadi SS. The Association of panton-valentine leukocidin and mecA genes in methicillin-resistant Staphylococcus aureus isolates from patients referred to educational hospitals in Ahvaz, Iran. Jundishapur J Microbiol. 2015; 8(8): e22021.
18. Lu SY, Chang FY, Cheng CC, Lee KD, Huang YC. Methicillin-resistant Staphylococcus aureus nasal colonization among adult patients visiting emergency department in a medical center in Taiwan. PLoS ONE. 2011; 6(6): e18620.
19. Shakeri F, Golalipour M, Rahimi Alang S, Ghaemi EA. Spa diversity among MRSA and MSSA strains of Staphylococcus aureus in north of Iran. Int J Microbiol. 2010; (2): 351397.
20. Ghaznavi-Rad E, Nor Shamsudin M, Sekawi Z, van Belkum A, Neela V. A simplified multiplex PCR assay for fast and easy discrimination of globally distributed staphylococcal cassette chromosome mec types in meticillin-resistant Staphylococcus aureus. Med Microbiol. 2010; 59(10): 1135-1139.
21. Ohadian Moghadam S, Havaei SA, Pourmand MR. Prevalence of methicillin-resistant Staphylococcus aureus carrying panton– valentine leukocidin gene in cutaneous infections in the city of Isfahan. J Med Bacteriol. 2012; 1(2): 9-16.
22. Zamani A, Sadeghian S, Ghaderkhani J, Alikhani MY, Najafimosleh M, Taghi Goodarzi M. Detection of methicillin-resistance (mec-A) gene in Staphylococcus aureus strains by PCR and determination of antibiotic susceptibility. Ann Microbiol. 2007; 57(2): 273-276.
23. Moradi N, Javadpour S, Karmostaji A. Reduced sensitivity of staphylococcus aureus to vancomycin. Hormozgan Uni Med Sciences. 2011; 15(3): 169-177.
24. Haghgoo S, Moaddab S, Rafi A. Study of antibiotic resistance pattern of Staphylococcus aureus strains isolated from blood cultures in Tabriz Shahid Madani Hospital. J Jundishapur. 2012; 3(2): 383-390.
25. Molla-abbaszadeh H, Mobayen H, Mirzaei H. Identification of pantonvalentine leukocidin (pvl) genes in Staphylococcus aureus isolated from ipatients of Emam Reza and Shohada Hospitals of Tabriz by real-time PCR. Iran J Med Microbiol. 2013; 6(4): 72-80.
26. Khosravi AD, Hoveizavi H, Farshadzadeh Z. The prevalence of genes encoding leukocidins in Staphylococcus aureus strains resistant and sensitive to methicillin isolated from burn patients in Taleghani Hospital, Ahvaz, Iran. Burns. 2012; 38(2): 247-251.
27. Otokunefor K, Sloan T, Kearns AM, James R. Molecular characterization and panton-valentine leucocidin typing of community-acquired methicillin-sensitive Staphylococcus aureus clinical isolates. J Clin Microbiol. 2012; 50(9): 3069-72.
28. Osman NAM, Intisar IE, Mohamed YM, El-Eragi AM, Eldirdery MM, Salih MA. Molecular study of panton-valentine leukocidin genes among Staphylococcus aureus clinical isolates in Khartoum State, Sudan. Am J Microbiol Res. 2015; 3(3): 107-111.