Metisilin dirençli Staphylococcus aureus'ların arbitrarily primed PCR ve plazmid profil analizi ile değerlendirilmesi

Bu çalışmada; hastanemizin Genel Cerrahi, Ortopedi ve Travmatoloji ile Plastik ve Rekonstrüktif Cerrahi kliniklerinde yatarak tedavi gören 30 hastadan Haziran-Aralık 1997 tarihleri arasında hastane infeksiyonu etkeni olarak izole edilen ve tanımlanan 30 metisiline dirençli Staphylococcus aureus (MRSA) izolatı "arbitrarily primed-polymerase chain reaction (AP-PCR)" ve plazmid profil analizi (PPA) yöntemleriyle moleküler olarak tiplendirilmiştir. Hastalardan infeksiyon etkeni olarak izole edilen 30 MRSA izolatı AP-PCR yöntemi ile bant paternlerine göre yedi grupta (1. grupta: 13,11. grupta: 10, 111. grupta: 2, IV. grupta: 2, V. grupta: I, VI. grupta: 1 ve VII. grupta: I), PPA yöntemi ile ise plazmid profiline göre üç grupta (A grubunda: 22, B grubunda: 3, C grubunda: 5) toplanmıştır. Çalışma sonucunda ilgili kliniklerdeki hastalardan elde edilen izolatların, özellikle 1. ve II. grup ile A grubunda (sırasıyla AP-PCR %77, PPA %73) yer aldığı belirlenmiş ve bu izolatların hastanemizde hastane infeksiyonlarına en sık neden olan kökenler olduğu saptanmıştır.

The evaluation of the Staphylococcus aureus strains by using arbitrarily primed-polymerase chain reaction and plasmid profile analysis techniques

A total of 30 methicillin resistant Staphylococcus aureus (MRSA) strains were isolated as nosocomial pathogens from patients hospitalized in General Surgery, Orthopedics and Reconstructive Surgery Departments from June 1997 through December 1997. These strains were evaluated by moleculer typing methods using arbitrarily primed-polymerase chain reaction (AP-PCR) and plasmid profile analysis (PPA). By using AP-PCR method, these isolates on the basis of their band patterns were divided into seven groups (I through VII) in which each group had reciprocal number of isolates by thirteen, ten, two, two, one, öneand one, respectively. By PPA, these isolates were grouped into three divisions as group A, B and C and each was represented by twentytwo, three and five isolates respectively. It was found that as nosocomial pathogens group I and II (77%) are frequently encountered for AP-PCR, while group A for PPA (73%) dominated.

Kaynakça

1. Musser JM, Kapur V. Clonal analysis of methicillin resistant Staphylococcus aureus strains from intercontinental sources: Association of the mec gene with divergent phylogenetic lineages implies dissemination by horizontal transfer and recombination. J Clin Microbiol 1992; 30:2058-63.,

2. Fang FC, Mc Clelland M, Donald GG, et al. Value of moleculer epidemiologic analysis in nosocomial methicillinresistant Staphylococcus aureus outbreak. JAMA 1993; 270:1323-8.

3. Tenover FC, Arbeit R, Archer G, et al. Comparision of traditional and moleculer methods of typing isolates of Stapylococcus aureus. J Clin Microbiol 1994;32:407- 15.

4. Struelens MJ, Deplano A, Godard C, Nicole M, Serruys E. Epidemiologic typing and delineation of genetic relatedness of methicillin-resistant Staphylococcus aureus by macrorestriction analysis of genomic DNA by using pulsed-field gel electrophoresis. J Clin Microbiol 1992; 30:2599-605.

5. Kumari PDN, Keer V, Hawkey PM, et al. Comparision and application of ribosome spacer DNA amplicon polymorphisms and pulsed-field gel electrophoresis for differentitation of methicillin-resistant Staphylococcus aureus strains. J Clin Microbiol 1997;37:881-5.

6. Saulnier P, Bourneix C, Prevost G, Andremont A. Random amplified polymorphic DNA assay is less discriminant than pulsed-field gel electrophoresis for typing strains of methicilin-resistant Staphylococcus aureus. J Clin Microbiol 1993;31:982-5.

7. Albay A, Yıldıran ŞT, Saraçlı MA, Kısa Ö, Güney Ç. Nozokomiyal MRSA suşlarının plazmid profillerinin araştırılması. Gülhane Tıp Dergisi 1999;41:134-7.

8. Otkun M, Akata F, Kocagöz S, ve ark. Metisiline dirençli Staphylococcus aureus’un kantitatif antibiyogram ve “Arbitrarily primed-PCR (AP-PCR)” yöntemleriyle epidemiyolojik sürveyansı. Hastane İnfeksiyonları Dergisi 1997;1:106-15.

9. Zuccareli AJ, Roy I, Harding GP, Couperus JJ. Diversity and stability of restriction enzyme profiles of plasmid DNA from methicillin-resistant Staphylococcus aureus. J Clin Microbiol 1990;28:97-102.

10. Collins JK, Smith JS, Kelly MT. Comparison of phage typing, plasmid mapping, and antibiotic resistance patterns as epidemiological markers in nosocomial outbreak of methicillin-resistant Staphylococcus aureus infections. Diagn Microbiol Infect Dıs 1984;2:239-45.

11. Oboyashi Y, Fujita J, Ichiyama S, et al. Investigation of nosocomical infection caused by arbekacin-resistant Staphylococcus aureus. Diagn Microbiol Infect Dis 1997;28:53-9.

12. Maslow JN, Mulligan ME, Arbeit RD. Molecular epidemiology: Applications of contemporary techniques to the typing of microorganisms. Clin Infect Dis 1993;17:153- 64.

13. Farmer JJ. Staphylococcus and Micrococcus. In: Kloos WE, Bannerman TL (eds). Manual of Clinical Microbiology. 7th ed, Washington: American Society for Microbiology, 1999:264-82.

14. Koneman EW, Allen SD, Janda WM, Schreckenberger PC, Winn WC. Color Atlas and Textbook of Diagnostic Microbiology. 5th ed, Philadelphia: Lippincott-Raven, 1997:539-66.

15. National Committee for Clinical Laboratory Standards. Performance Standards for Antimicrobial Susceptibility Tests. Ninth Informational Supplement. M100-S9, Vol. 19, No: 1, Villanova PA. NCCLS, 1999.

16. Ünal S, Hopkins J, Flokowitsch JE, Ernie WU CY, Preston DA, Skatrud PL. Detection of methicillin-resistant staphylococci by using the polymerase chain reaction. J Clin Microbiol 1992;30:1685-91.

17. Welsh J, McClelland M. Characterization of Pathogenic Microorganisms by Genomic Fingerprinting Using Arbitrarily Primed-PCR. In: Persing DH, Smith TF, Tenover FC, White TJ (eds). Diagnostic Molecular Microbiology, Principles and Applications. American Society for Microbiology, Washington DC. 1993 Mayo Foundation Rochester, 595-62.

18. Wildemauwe C, Godard C, Vanhoof R, Van Bossuyt E, Hannecart-Pokorni E. Changes in major populations of methicillin-resistant Staphylococcus aureus in Belgium. J Hosp Infect 1996;34:197-203.

19. Bialkowska-Hobrzanska H, Jaskot D, Hammerberg O. Evaluation of restriction endonuclease fingerprint of chromosomal DNA and plasmid profile analysis for characterization of multiresistant coagulase-negative staphylococci in bacteremic neonates. J Clin Microbiol 1990; 28:269-75.

20. Wachsmuth K. Molecular epidemiology of bacterial infections: Examples of methodology and of investigations of outbreaks. Review Infect Dis 1986;8:682-92.

21. Einstein BI. New moleculer techniques for microbial epidemiology and the diagnosis of infectious diseases. J Infect Dis 1990;161:595-602.

22. Hartstein AI, Morthland VH, Eng S, Archer GL, Schoenknecht FD, Rashad AL. Restriction enzyme analysis of plasmid Staphylococcus aureus blood culture isolates. J Clin Microbiol 1989;27:1874-9.

23. Tricinski K, Hryniewicz W, Kluytmans J, et al. Simultaneous persistence of methicillin-resistant and methicillinsusceptible clones of a warsaw hospital. J Hosp Infect 1997;36:291-303.

Kaynak Göster