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  1. Project presentation on “ Detection of Staphylococcal carriage state among Nursing Staff” Presented by, Ayesha amreen 2nd M.sc., 4th semester Microbiology M.S.C.W Mysore

  2. Staphylococci - most commenest ubiquitous bacteria Staphylococcus aureus - leading pathogen causing nosocomial & also community acquired infections. Indiscriminate use of antibiotics - Drug resistance Penicillin - Very effective medication to treat Staphylococcal infections Penicillin resistance - Penicillinase inactivated the antibiotic Discovery of new antibiotic Methicillin in 1959 Methicillin resistance Staphylococcus aureus(MRSA) - 'Superbug' Mechanism of drug resistance - alteration in the binding sites and production of enzymes Methicillin resistance was mediated by penicillin binding protein[PBP2a], Encoded by the mec A gene situated on the mobile genetic element, Staphyloccal cassette chromosome mec (SCC mec) Introduction

  3. Resistant strains developed in hospitals are being increasingly detected as the causative agents of Community Acquired & Nosocomial wound infections. Hand carriage of Staphylococcus aureus in health care workers is the commonest mode of transmission to patients during cleaning of wounds. Healthy carrier state of Staphylococcus aureus - A major problem

  4. To screen the nursing staff of high risk areas such as ICU, OT, Emergency ward etc., for Staphylococcal colonization in the throat & on the hands. To detect the MRSA carriage rate among the Nursing staff. To study the antibiogram of the Staphylococcal isolates. Aims and objectives

  5. Materials and methods Swab used to collect sample Collection of finger and throat swab

  6. Materials and methods Title Culturing on Mannitol salt agar medium Pink colonies Yellow colonies Identification of S.aureus on MSA Tube Coagulase test

  7. Materials and methods Oxacillin resistance salt screening agar test Antibiotic Susceptibility Test

  8. Results SEX AND WARD DISTRIBUTION OF NURSING STAFF Out of110 nursing staffs, 18 were males & 92 were females. Majority ofthe nursing staff were from Dialysis unit & Major OT. STAPHYLOCOCCAL GROWTH FROM THROAT SWAB Among110 nursing staff screened,14 isolates were isolated from throat swabsof nursing staff, of the 14 isolates, 11 isolates yielded the growth ofcoagulase positive staphylococci [COPS]& 3 isolates yielded thegrowth of coagulase negative staphylococci [CONS] . out of 11 isolates,significant growth of coagulase positive staphylococci was observedamong 5 nursing staff of Dialysis unit

  9. STAPHYLOCOCCAL GROWTH FROM FINGER SWAB Outof 110 nursing staff screened,14 isolates were isolated from fingerswabs of nursing staff, of the 14 isolates, 3 isolates yielded thegrowth of coagulase positive staphylococci [COPS]& 11 isolatesyielded the growth of coagulase negative staphylococci [CONS]. Amongthe 3 isolates all the 3 were from Dialysis unit & significantgrowth of coagulase negative staphylococci was observed among 4 nursingstaff of Dialysis unit & Major OT Pink coloured coagulase negative Staphylococcal colonies Yellow coloured Staphylococcus aureus colonies

  10. DETECTION OF OXACILLIN RESISTANCE BY USING OXACILLIN RESISTANT SCREENING AGAR IN THROAT SWAB Among110 nursing staff screened, 11 isolates yielded the growth of coagulasepositive staphylococci in the throat swabs& all the 11 isolateswere carriers of Methicillin Sensitive Staphylococcus aureus[MSSA] & none of them carried Methicillin Resistant Staphylococcus aureus [MRSA] 3 isolates yielded the growth of coagulase negative staphylococci . Ofthe 3 isolates 1 was carrier of Methicillin Sensitive- coagulasenegative Staphylococcus aureus [MS-CONS] & 2 of them carriedMethicillin Resistant- coagulase negative Staphylococcus aureus[MR-CONS]

  11. DETECTION OF OXACILLIN RESISTANCE BY USING OXACILLIN RESISTANT SCREENING AGAR IN FINGER SWAB Outof 110 nursing staff screened, 3 isolates yielded the growth ofcoagulase positive staphylococci in the finger swabs& all the 3isolates were carriers of Methicillin Sensitive Staphylococcus aureus[MSSA] & none of them carried Methicillin Resistant Staphylococcus aureus [MRSA] 11 isolates yielded the growth of coagulase negative staphylococci . Ofthe 11 isolates 8 were carriers of Methicillin Sensitive- coagulasenegative Staphylococcus aureus [MS-CONS] & 3 of them carriedMethicillin Resistant- coagulase negative Staphylococcus aureus[MR-CONS]

  12. COMPARISION OF DISC DIFFUSION WITH ORSA METHOD FOR DETECTION OF MRSA IN THROAT SWAB Among 8 coagulase positive S taphylococci, 7 of them were sensitive by both disc diffusion & ORSA methods.Only 1 strain showed resistance to oxacillin by disc diffusion method ,but on ORSA none of the strain was resistant COMPARISION OF DISC DIFFUSION WITH ORSA METHOD FOR DETECTION OF MRSA IN FINGER SWAB Among 3coagulase positive staphylococci , all the 3 strains were sensitive byboth disc diffusion & ORSA methods & none of the strains wereresistant to oxacillin by both methods.

  13. blue colony - MRSA 1. Result plate 2. standard plate 1. Oxacillin Resistant Salt Screening agar plate showing negative result for Methicillin Resistant Staphylococcus aureus. 2. Oxacillin Resistant Salt Screening agar plate showing Positive result for Methicillin Resistant Staphylococcus aureus.

  14. Conclusion Of the 110 participants, from 12 wards 18 were males & 92 were females. Of 110 nursing staffs screened ,the Staphylococcal carriage rate in both throat & on hand was 12.7% .None of them werecarriers of MRSA which was identified on the basis of disc diffusion& Oxacillin Screen Agar test. All isolates were sensitive toVancomycin. A considerably low Staphylococcal carriage rate noticed in our study , lowers the risk of carriage of MRSA strain also. Staphylococcus aureus throat & hand carriers nurses should be identified & trained topractice infection control measures. Nurses who acquire MRSA inhospital also transmit the organism to their hosehold eventuallyspreading such Nosocomially acquired Multidrug Resistant bacteria inthe community , therefore, such carriage study should be conducted at aregular basis in all health sectors followed by the treatment of theidentified carriers.

  15. References 1. Barber, M. Methicillin-resistant staphylococci. J Clin Pathol 1961; 14:385. 2. Benner, EJ, Kayser, FH. Growing clinical significance of methcillin-resistant Staphylococcus aureus. Lancet 1968; 2:741. 3. Abdelkarim Waness. Revisiting methicillin-resistant Staphylococcus aureus infections. Year : 2010 | Volume : 2 | Issue : 1 | Page : 49-56 4. Practical Medical Microbiology. 14th edition. Mackie & Mc.Cartney.J.Gerald collee, Andrew G.Fraser, Barrie P.Marmion, Antony simmons. Churchill Livingstone. pp. 245-258.

  16. 5. National Committee for Clinical Laboratory Standards: Methods for dilution antimicrobial susceptibility testing for bacterial that grow aerobically. Approved standard M7-A5. 2000, National Committee for Clinical Laboratory Standards, Wayne, Pa 5th edition. http://www.netwellness.org/ http://www.gdargaud.net http://www.nlm.nih.gov/ http://www.calgarylabservices.com http://www.precisionliftinc.com/ http://www.inside-hospitals.co.uk/

  17. THANK YOU

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