원유시료 중 메티실린 내성 황색포도알균의 분포 및 내성 유전자 특성 분석

Abstract

원유시료에서 분리한 methicillin 내성 황색포도알균의 분표율과 내성유전자의 분석을 분자수준에서의 실시하였다. 총 698종의 원유시료에서 286개의 포도알균속 세균을 Staphylococcus Medium 110을 이용하여 분리 후 Vitek 2 system을 이용하여 동정한 결과 94개의 황색포도알균을 확인하였다. 94개의 황색포도알균 중 총 7개의 균에서 methicillin 내성 mecA 유전자가 존재함을 확인하였다. Methicillin-resistant Staphylococcus aureus (MRSA)로 확인된 7 균주에 대해 수종의 항생제에 대한 감수성 시험, staphylococcal cassette chromosome mec (SCCmec) typing, 및 병독소유전자인 Panton-Valentine Leukocidin (pvl)유전자를 분석하였다. 그 결과 7종의 mecA 양성 MRSA 모두 ampicillin과 oxacillin에 내성이었으며 teicoplanin, vancomycin 및 ciprofloxacin에는 모두 감수성임을 확인하였다. 한편 SCCmec typing결과 7균 중 1균주는 SCCmec type II로 확인되었고 6균주는 SCCmec type IV로 확인되었으며 병독소 유전자인 pvl유전자는 검출되지 않았다. This study was investigated to determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) isolated from raw milk samples and to further study on the molecular characteristics of the MRSA isolates. Using Staphylococcus Medium 110, Staphylococcus spp. were isolated from raw milk samples and further identification was carried by Vitek2 system. Minimum inhibitory concentrations (MICs) of antibiotics were conducted by serial dilution method according to the Clinical Laboratory Standards Institute (CLSI) guideline. For the detection of resistance genes and molecular characterization, PCR reaction was performed by gene specific primers and followed by DNA sequencing. Of the 698 milk samples, 94 Staphylococcus aureus (S. aureus) were identified (94 S. aureus/286 Staphylococcus spp.). Of the 94 S. aureus, seven isolates have mecA, a methicillin resistant gene. mecA positive seven isolates were then characterized by staphylococcal cassette chromosome mec (SCCmec) typing, and Panton-Valentine Leukocidin (pvl) gene using PCR. All of mecA positive isolates were resistant to ampicillin and oxacillin, but sensitive to teicoplanin, vancomycin and ciprofloxacin. One of seven isolates was SCCmec type II and six isolates were type IV and all seven isolates were pvl gene negative.