초록 ▼

5. 최종 연구결과 요약
○ 세부과제명: 식중독 유발 병원체 SNP 패턴을 이용한 multiplex PNA chip 개발연구
- 식중독균 3종 증균배지에 따른 식중독균의 증식 결과 확인
- 식중독균 3종 증균 배지와 배양시간에 따른 Multiplex PCR과 PNA칩
- 축산물의 가공기준 및 성분규격에 따른 식육 PNA chip 적용실험
- 닭고기, 돼지고기, 쇠고기 등의 식육에서의 PNA 칩을 이용한 식중독균 검출시험
- 가축에서 분리된 야외분리주에 대한 PNA칩 적용시험
• Clostridi

5. 최종 연구결과 요약
○ 세부과제명: 식중독 유발 병원체 SNP 패턴을 이용한 multiplex PNA chip 개발연구
- 식중독균 3종 증균배지에 따른 식중독균의 증식 결과 확인
- 식중독균 3종 증균 배지와 배양시간에 따른 Multiplex PCR과 PNA칩
- 축산물의 가공기준 및 성분규격에 따른 식육 PNA chip 적용실험
- 닭고기, 돼지고기, 쇠고기 등의 식육에서의 PNA 칩을 이용한 식중독균 검출시험
- 가축에서 분리된 야외분리주에 대한 PNA칩 적용시험
• Clostridium perfingens의 검출능 조사 (A, B, C, D, E 형)
- 축산 식품의 PNA칩 적용시험 (도축장/도계장)
• 소고기, 닭고기 식육에서 식중독균 검출조사
- 축산가공품의 PNA 칩 적용시험
• 시중에 유통되고 있는 우유 소세지 등 축산가공품 98점을 수거하여 식중독균 검출 조사 ○세부과제명: Clostridium spp 검출용 poultry ELISA 및 백신개발
- 재조합 NetB항원과 ISA71 adjuvant 혼합 백신은 체중감소억제, Lesion Score 감소 및 Cytokine생산 억제함
- 재조합 PFO항원과 ISA71 adjuvant 혼합 백신 경우 체중증가, Lesion Score 감소 및 IL-8생산 억제함
- Clostridium perfringens 독소에 의한 NE유발에 대한 백신후보군으로 재조합 NetB와 PFO항원을 선발함

Abstract ▼

Food poisoning by Salmonella sp, Camphylobacter sp, C.perfringens causes diarrhea, bloody stools and abdominal pain in humans, and may lead to severe kidney failure or brain injury.
Anovel peptide nucleic acid (PNA) array was developed to identify Salmonella sp, Camphylobacter sp, C.perfringens q

Food poisoning by Salmonella sp, Camphylobacter sp, C.perfringens causes diarrhea, bloody stools and abdominal pain in humans, and may lead to severe kidney failure or brain injury.
Anovel peptide nucleic acid (PNA) array was developed to identify Salmonella sp, Camphylobacter sp, C.perfringens quickly and accurately. This multiplex PNA array exhibited high specificity, significantly detecting Salmonella sp, Camphylobacter sp, C.perfringens strains without cross-reacting with other bacterial strains. This multiuplex PNA array could be a valuable technique for high-through put screening of pathogens that cause food poisoning in humans.
This study was performed to compare four Clostridium perfringens recombinant proteins as vaccine candidates using the Montanide™ ISA 71 VG adjuvant in an experimental model of necrotic enteritis. Broiler chickens were immunized subcutaneously with purified clostridial recombinant NetB toxin, pyruvate: ferredoxin oxidoreductase (PFO), α-toxin, or elongation factor-Tu (EF-Tu), or with vehicle control, in conjunction with ISA 71 VG, and intestinal lesion scores, body weight gains, NetB toxin and PFO antibody levels, and proinflammatory cytokine andchemokine levels were measured as outcomes of protection following oral co-infection with C. perfringens and Eimeria maxima. Birds immunized with all recombinant proteins plus ISA 71 VG showed significantly reduced gut lesions compared with the ISA 71 VG-only group. Birds immunized with NetB toxin or PFO plus ISA 71 VG exhibited significantly increased body weight gains compared with the ISA 71 VG alone group. Greater NetB toxin antibody titers were observed in the NetB/ISA 71 VG group, and greater PFO antibody titers were evident in the PFO/ISA 71 VG group, each compared with the other three vaccine/adjuvant groups. Finally, decreased levels of gene transcripts encoding interleukin-8, tumor necrosis factor superfamily 15, and LPS-induced TNF-α factor were observed in the intestinal lymphocytes of chickens immunized with NetB toxin, PFO, α-toxin, and/or EF-Tu in the presence of ISA 71 VG compared with ISA 71 VG alone. All parameters evaluated were equal in co-infected chickens given ISA 71 VG alone compared with infected/adjuvant-free birds, indicating that the adjuvant itself did not have a disease protective effect. These results suggest that vaccination with clostridial recombinantproteins, particularly NetB toxin or PFO, in combination with ISA 71 VG enhances protective immunity against experimental necrotic enteritis in broiler chickens.

목차 Contents

• 총괄요약표 ... 1
• 연구과제 최종결과보고서 ... 2
• I. 연구배경 및 목표 ... 2
• II. 연구방법 및 수행전략 ... 5
• III. 연구결과 ... 9
• IV. 연구결과요약 ... 24
• V. 고찰 ... 24
• VI. 연구결과 활용실적 및 계획 ... 25
• VII. 참고문헌 ... 25
• X. 영문초록 ... 27
• 끝페이지 ... 27