세균성 피부염 개에서 분리된 Staphylococcus pseudintermedius에서 항생제 감수성 검사와 내성 유전자 획득의 비교 Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis원문보기
세균성 피부염은 항생제 치료가 필요한 흔한 질환이다. 최근 항생제 내성 세균이 전 세계적으로 증가하고 있는 추세이다. 본 연구에서는 개 피부염 병변에서 분리한 S. pseudintermedius의 항생제 내성 유전자 발현을 조사하고, 이를 디스크 감수성 시험결과와 비교하였다. 총 7개의 S. pseudintermedius 균주가 연구에 포함되었다. 세균 동정은 16S rDNA 염기 서열 분석으로 실시하였다. 조사 결과 분리한 S. pseudintermedius 모두가 1개 이상의 항생제 내성유전자(mecA, blaZ and aac[6']/aph[2"])를 보유하였다. 전체 분리균주가 blaZ 유전자에 양성을 보였으나, 두 개 균주만이 amoxicillin/clavulanate에 내성을 나타냈다. Gentamicin 내성을 나타낸 5개 균주 중, 1개는 aac(6')/aph(2") 유전자에 대한 PCR 검사에서 음성을 나타냈다. 이 결과는 내성 유전자 검출 PCR 기법과 디스크 감수성 시험 기법이 항생제 내성을 검사하는 데에 있어 상호보완적인 관계를 가짐을 제시한다.
세균성 피부염은 항생제 치료가 필요한 흔한 질환이다. 최근 항생제 내성 세균이 전 세계적으로 증가하고 있는 추세이다. 본 연구에서는 개 피부염 병변에서 분리한 S. pseudintermedius의 항생제 내성 유전자 발현을 조사하고, 이를 디스크 감수성 시험결과와 비교하였다. 총 7개의 S. pseudintermedius 균주가 연구에 포함되었다. 세균 동정은 16S rDNA 염기 서열 분석으로 실시하였다. 조사 결과 분리한 S. pseudintermedius 모두가 1개 이상의 항생제 내성유전자(mecA, blaZ and aac[6']/aph[2"])를 보유하였다. 전체 분리균주가 blaZ 유전자에 양성을 보였으나, 두 개 균주만이 amoxicillin/clavulanate에 내성을 나타냈다. Gentamicin 내성을 나타낸 5개 균주 중, 1개는 aac(6')/aph(2") 유전자에 대한 PCR 검사에서 음성을 나타냈다. 이 결과는 내성 유전자 검출 PCR 기법과 디스크 감수성 시험 기법이 항생제 내성을 검사하는 데에 있어 상호보완적인 관계를 가짐을 제시한다.
Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs,...
Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.
Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.
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제안 방법
, inducible clindamycin resistance usually produces false-negative result in conventional disc diffusion tests) (3). In this study, we examined antibiotic resistance profiles of the S. pseudintermedius isolates from canine pyoderma in South Korea by using standard disc diffusion test and resistant gene-targeted PCR and then compared the results between two test methods.
To identify S. pseudintermedius, extracted bacterial genomic DNA was amplified using a pair of 27F (5’-AGAGTTTGATCCTGGCTCAG-3’) and 1492R (5’-GGTTACCTTGTTACGACTT-3’) for the 16S ribosomal RNA gene (16S rDNA).
대상 데이터
A total of 56 canine patients with bacterial dermatitis was included in the study. The sample was collected by cotton swab on the skin lesions.
Among the bacterial samples (n = 56), seven S. pseudintermedius were isolated and used in the study. Disc diffusion test showed that all isolates had resistance to more than one antibiotic (Table 2).
pseudintermedius, antibiotic susceptibility testing was performed by the Kirby-Bauer disc diffusion test. For the test, 3 antibiotics were selected: oxacillin, gentamicin, and amoxicillin/clavulanate (BBLTM Sensi-DiscTM Susceptibility test disc, BD, NJ, USA). The interpretation of the test results were made based on the CLSI guideline for veterinary antimicrobial susceptibility testing for pathogens of animal origin (CLSI, 2008).
이론/모형
For the test, 3 antibiotics were selected: oxacillin, gentamicin, and amoxicillin/clavulanate (BBLTM Sensi-DiscTM Susceptibility test disc, BD, NJ, USA). The interpretation of the test results were made based on the CLSI guideline for veterinary antimicrobial susceptibility testing for pathogens of animal origin (CLSI, 2008).
All amplicons were purified and sent for DNA nucleotide sequence analysis (SolGent, Daejeon, Korea). The sequence homology of deduced nucleotide sequences was analyzed with the BLAST search program (National Center for Biotechnology Information, NCBI) based on the interpretative criteria for identifying bacteria and fungi by DNA target sequence (The Clinical and Laboratory Standards Institute guideline MM-18A, CLSI, Wayne, PA, USA).
To examine the antibiotic resistance of the isolated S. pseudintermedius, antibiotic susceptibility testing was performed by the Kirby-Bauer disc diffusion test. For the test, 3 antibiotics were selected: oxacillin, gentamicin, and amoxicillin/clavulanate (BBLTM Sensi-DiscTM Susceptibility test disc, BD, NJ, USA).
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