생물막형성 장내세균의 Curli 및 Cellulose 세포외 바탕질 분석 = Biofilm Forming Ability and Production of Curli and Cellulose in Clinical Isolates of Enterobacteriaceae

국문 초록 (Abstract)

본 연구에서는 병원 감염의 주요 원인균으로 생물막 관련 감염증에서 흔히 분리되고 있는 Citrobacter, Enterobacter 및 Serratia 등의 임상분리 장내세균 22주를 대상으로 생물막 형성능을 조사하고, 생물막의 세포외 바탕질의 구성 성분을 알아보기 위하여 Congo-red 한천배지상의 집락 형상과 calcofluor 염색을 시행하였다. 또한 curli 생성 오페론인 csgBA(C) 유전자의 유무 확인과 csgA 유전자의 염기서열을 규명하였다. $37^{\circ}C$에서 24시간 배양 후 생물막 형성능 분석에서는 2주의 S. marcescens를 제외한 나머지 20주는 모두 생물막 형성능이 있는 것으로, $28^{\circ}C$에서 48시간 배양 후 분석에서는 22주 모두 생물막 형성능이 있는 것으로 확인되었다. Congo-red 한천배지에서의 균집락 형상을 조사한 결과 균속에 따라서 균 집락의 표현형상이 다르게 나타났으며, 동일 균속내에서도 균 집락의 표현형상과 색 농도의 차이가 관찰되어 세포외 바탕질의 주요 성분 및 생성량의 차이가 있음을 시사하였다. 1주의 C. freundii와 4주의 E. cloacae에서 csgBA(C) 유전자 양성을 나타내었다. 염기서열 분석결과, E. cloacae의 csgA는 E. coli의 csgA와 80.9%, E. sakazakii의 csgA와 75.7%, 그리고 C. freundii의 csgA와 67.8%의 상동성이 있는 것으로 나타났다. E. cloacae의 경우 Congo-red 염색의 결과와 curli 유전자 검색 결과가 일치하였을 뿐만 아니라, csgA 유전자를 보유하지 않은 균주의 생물막 형성능이 csgA 유전자를 보유한 균주에 비해 현저하게 낮은 것으로 나타나, curli가 E. cloacae의 생물막 세포외 바탕질 구성성분의 주요 요소일 것으로 추정되었다.

다국어 초록 (Multilingual Abstract)

In this study, 22 clinical isolates of Enterobacteriaceae including Citrobacterfreundii (6 strains), Enterobacter cloacae (5 strains), Enterobacter aerogenes (3 strains), Serratia marcescens (7 strains) and Pantoea spp. (1 strain) were investigated for the biofilm forming ability and biosynthesis of curli and cellulose. Biofilm forming ability was the highest among the isolates of E. cloacae and the lowest among the isolates of E. aerogenes. The expression of the biofilm-forming extracellular matrix components, cellulose and curli fimbriae, was examined by Congo-red (CR) staining and calcofluor staining methods. PCR screening for the presence of curli gene (csgA) revealed that 4 strains of E. cloacae and 1 strain of C. freundii carried the csgA, showing a good correlation between the phenotypic detection of curli fimbriae by CR staining method and the genotypic detection of curli gene by PCR in E. cloacae.

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