Seok Ki Lee, Dong Hwan Choi, Hyou-Arm Joung, Nae-Rym Lee, Jun Hyoung Ahn, Bong Hyun Chung, Yong-Beom Shin, Min-Gon Kim
언어
영어(ENG)
URL
https://www.earticle.net/Article/A98636
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원문정보
초록
영어
Rapid detection of food-borne bacterial pathogens is desperately needed in clinical, food safety and. environmental field. In this study, we developed a rational method to evaluate the diagnostic ability by using a fluorescence labeled colony-PCR (FLC-PCR) that is possible for simultaneous detection of 16S ribosomal DNA of multiple bacterial species. The diagnostic ability of the Fluorescent (Cyanine 5) labeled colony polymerase chain reaction (FLC-PCR) based DNA microarray method was evaluated. The PCR frgments were carried out successfully, and these genes were differentiated by the heterologous region of their PCR products in same region of bacterial 16s rDNA sequence. The 8 species-specific capture probes (Eco2, Eco3, Lisn2, Lis3, Clo3, StaA, Vib1, and Vc1, 5'-Amine labelled) were designed from 16S rDNA genes encoding food borne bacterial genomic DNA of different six pathogens (Escherichia coli O157:H7, Vibrio cholera, Vibrio parahaemolyticus, Listeria monocytogenes, Clostridium perfringens, and Staphylococcus aureus). The FLC-PCR frgments (302 bp) of 6 pathogens were hybridized 1 hour (60℃) to immobilize speciesspecific and universal probes on microarray. The fragment sizes of PCR products were identical to the sizes calculated from nucleotide sequences of 16s rRNA deposited in EMBL, GenBank, and/or DDBJ databases, and the sequences were also identical. In conclusions, the method and newly designed primers applied in this work can differentiate the Pathogen’s 16s rRNA sequence very fast (< 3 hours) and effectively, without liquid cultivation and purified DNA.
저자
Seok Ki Lee [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Dong Hwan Choi [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Hyou-Arm Joung [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Nae-Rym Lee [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Jun Hyoung Ahn [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Bong Hyun Chung [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Yong-Beom Shin [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
Min-Gon Kim [ BioNanotechnology Research Center,KRIBB, Daejeon 305-806, South Korea ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동