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병·의원에서 분리한 메티실린 내성 황색포도알균에서 SCCmec 유형과 Aminoglycoside 변형효소 유전자의 분포

Distribution of Genes Encoding Aminoglycoside Modifying Enzymes and Type Staphylococcal Chromosomal Cassette mec in Methicillin-resistant Staphylococcus aureus from Non-tertiary Hospitals

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초록

목 적 : 대부분의 AME 효소 유전자는 transposon이나 plasmid 상태로 전달되며 staphylococcal chromosomal cassette mec (SCCmec) 내에도 삽입되어 있다. 최근 MRSA의 SCCmec 유형에 따라 다양한 내성유전자를 가지는 것으로 보고되고 있다. 따라서 1, 2차 의료기관에서 분리된 황색포도알균의 항생제 내성률을 조사하고 MRSA 에서 SCCmec type에 따른 AME 효소 유전자의 분포를 확인하였다. 재료 및 방법 : 2004년 1, 2차 의료기관에서 분리된 황색포도알균 640주를 대상으로 methicillin의 내성유전자 mecA와 AME 효소 유전자 aac(6')-aph(2"), aph(3')-IIIa, ant(4')-Ia 등을 multiplex PCR 방법을 이용하여 확인하였다. 항생제 감수성 시험과 MIC 값은 한천배지 희석법으로 확인하였고 SCCmec type도 결정하였다. 결 과 : 640주의 황색포도알균 중 MRSA율은 39.7%였으며 MRSA 분리주 모두 mecA 유전자가 검출되었다. Aminoglycoside 중 kanamycin, tobramycin에 대한 내성률은 98.1%이었고, gentamicin 68.7%, amikacin 30.8%, netilomicin 2.8%이었으며 vancomycin에는 모두 감수성이었다. Oxacillin MIC50과 MIC90은 각각 128 µg/mL, 256 µg/mL이었으며 254주의 MRSA 중 214주를 선별하여 AME 효소 유전자를 확인한 결과 aph(3')-IIIa 13.1%, aac(6')-aph(2") 77.1%, ant(4)-Ia 53.3%이었으며 SCCmec type에 따라서는 type II, type III, type IV가 각각 50.9%, 16.4%, 32.7%였다. SCCmec type에 따른 AME 효소 유전자의 분포는 SCCmec type II에서는 aac(6')-aph(2")와 aac(6')-aph(2")/ant(4')-Ia가 각각 49.5%, 36.7%에서 검출되었고 type III에서는 aph(3')-IIIa/aac(6')-aph(2")가 60%, aac(6')-aph(2")가 31.4%, type IV에서는 aac(6')- aph(2")/ant(4')-Ia와 ant(4')-Ia가 각각 41.4%, 50%로 나타났다. 결 론 : 1, 2차 의료기관에서 분리한 황색포도알균 분리주의 메티실린 내성률은 39.7%이었다. MRSA 분리주 중 SCCmec type II와 III에서는 aac(6')-aph(2")가, SCCmec type IV에서는 ant(4')-Ia가 90% 이상 검출되어 SCCmec type과 AME 효소 유전자 분포와는 연관성이 있는 것으로 나타났다.

Abstract

Background : Many genes encoding aminoglycoside modifying enzymes (AMEs) on transposon or plasmid were transferred from one strain to another strain and inserted into a staphylococcal chromosomal cassette mec (SCCmec). There are very diverse subtypes in SCCmec type to the insertion of resistant genes. Therefore, we researched the resistance rates of antibiotics and distribution of AME genes according to SCCmec type in MRSA strains. Materials and Methods : We isolated 640 Staphylococcus aureus from non-tertiary hospitals in 2004, detected mecA, aac(6')-aph(2"), aph(3')-IIIa, and ant(4')-Ia using the multiplex PCR method, tested antibacterial susceptibility disk diffusion and minimal inhibitory concentration, and determined SCCmec type. Results : Of 640 S. aureus isolates, MRSA rate was 39.7% and all MRSA isolates carried mecA gene. Among 214 MRSA selected, aminoglycoside-resistant rates were 98.1% in kanamycin and tobramycin, 68.7% in gentamicin, 30.8% in amikacin, and 2.8% in netilmicin. The detection rates for aac(6')-aph(2"), aph(3')-IIIa, and ant(4')-Ia were 77.1%, 13.1%, and 53.3%, respectively. Also, SCCmec type was 50.9% in SCCmec type II, 16.4% in type III, and 32.7% in type IV. The genes encoding AMEs were distributed aac(6')-aph(2") (49.5%) and aac(6')-aph(2")/ant(4')-Ia (36.7%) in SCCmec type II, aph(3')-IIIa/aac(6')-aph(2") (60%) and aac(6')-aph(2") (31.4%) in type III, and aac(6')-aph(2")/ant(4')-Ia (41.4%) and ant(4')-Ia (50%) in type IV. Conclusion : 39.7% of S. aureus isolated from non-tertiary hospitals was resistant to methicillin. More than 90% of MRSA isolates were detected aac(6')-aph(2") in SCCmec type II and III, and ant(4')-Ia in type IV. With these results, the genes encoding AMEs may be closed related to SCCmec type.

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