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Listeria 속균 신속 검출을 위한 면역크로마토그라피 킷트 개발
정병열,정석찬,김종만,Jung, Byeong-yeal,Jung, Suk-chan,Kim, Jong-man 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.2
We developed the one-step strip based on an immunochromatographic (IC) assay for the rapid detection of Listeria spp. Genus-specific monoclonal antibody to flagella of L. monocytogenes was conjugated with 40 nm colloidal gold particles which were prepared in our laboratory. The specificity of the IC strip was tested with pure cultured bacteria. All strains of the genus of Listeria spp. yielded positive reactions and 12 strains of non-Listeria were negative, resulting in a specificity of 100%. L. monocytogenes was artificially inoculated in raw pork macerated with listeria enrichment broth. And then it was 10-fold diluted from $8.7{\times}10^6$ to 8.7 CFU/ml. L. monocytogenes could be detected at a minimum of $8.7{\times}10^5CFU/ml$ before enrichment, $8.7{\times}10^2CFU/ml$ after primary enrichment and 8.7 CFU/ml after secondary enrichment, respectively. These results indicated that the IC strip exhibited high specificity and sensitivity in the detection of Listeria spp.
면역크로마토그라피 기법을 이용한 Salmonella 속균 신속 검출킷트 개발
정병열,정석찬,Jung, Byeong-yeal,Jung, Suk-chan 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.2
An immunochromatographic (IC) strip for the rapid detection of Salmonella spp. in the enriched sample was developed. Affinity purified Salmonella polyclonal antibody was conjugated with 40 nm colloidal gold particles which were prepared by citrate method in our laboratory. The antigen-antibody-gold complex was captured by Salmonella antibody attached to test line of nitrocellulose membrane during the capillary migration of sample. Specificity of the IC strip was calculated to be 100% (12/12) and sensitivity was 97.6% (41/42) in the test with pure cultured bacteria. Salmonella was artificially inoculated into raw pork macerated with enrichment broth. And then it was 10-fold diluted from $5.2{\times}10^{8}CFU/ml$ to 5.2 CFU/ml. The IC strip could detect $5.2{\times}10^{6}CFU/ml$ before enrichment. However, the lowest limit of detection was 5.2 CFU/ml after overnight incubation. The results indicated that the IC assay was a rapid, economical and simple method with high specificity and sensitivity for the detection of Salmonella spp. without using any equipment.
정병열,전용수,Jeong, Byeong-yeal,Jeon, Yong-soo 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.3
The objective of this study was to determine the prevalence of pneumonic bacteria in lungs and nasal swabs of cattle with respiratory diseases. From 95 pneumonic lungs of slaughtered cattle, 41 (43.2%) positive lungs were yielded with 54 pneumonic bacteria, which corresponded to P. multocida (n = 34), A. pyogenes (n = 14) and P. haemolytica (n = 6). One-hundred sixty seven pneumonic bacteria were isolated from 195 nasal swabs in calves, 64.7% (108 isolates) belonged to P. multocida, 16.2% to A. pyogenes, 13.8% to P. haemolytica and 5.4% to H. somnus. Fifty percents (n = 6) of isolates from pneumonic lungs of calves were identified as P. multocida. All isolates of P. multocida belonged to type A according to hyaluronidase test. Antimicrobial susceptibility tests showed that most isolates of P. multocida and P. haemolytica were sensitive to amoxicillin/clavulanic acid, cephalothin, ciprofloxacin, enrofloxacin, fluorophenicol and norfloxacin. The emergence of antimicrobial resistant Pasteurella spp. observed in this study, however, might limit such application. According to histopathological examination, pneumonia by mycoplasma or/and bacteria accounted for 92.8% among 69 pneumonic lungs of slaughtered cattle.
가축의 장내용물에서 Listeria 속균의 분포도 조사
정병열,임현숙,김봉환 한국수의공중보건학회 2003 예방수의학회지 Vol.27 No.1
We surveyed the prevalence of Listeria spp. in cecal contents of pig and chicken collected from abattoirs. only 4 L. lnnocua isolates were detected in a total of 100 pig samples. From the 100 chicken samples, 47 were confirmed positive for Listeria spp. Twenty eight of these 47 positive samples contained only L. lnnocua; 15 contained both L. monocytogenes and L. lnnocua; 4 contained only L. mnocytogenes. No other Listeria spp. were detected in this study. Listeria spp. were detected in 471115 (40.9%) blackened Fraser broth samples and in 4/85 (4.7%) samples in which Fraser broth did not blacken. Fourteen (73.7%) L. monocytogenes isolates were recovered after 48 h, not 24 h, Fraser broth secondary enrichment.
돼지 폐렴병소에서 분리한 Actinobacillus pleuropneumoniae의 특성에 관한 연구
정병열,조길재,김봉환,조광현,Jung, Byeong-yeal,Cho, Gil-jae,Kim, Bong-hwan,Cho, Kwang-hyun 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.1
The present study was conducted to investigate the biochemical and serologic characteristic of Actinobacillus pleuropneumoniae isolated from pneumonic lungs of pigs during the period from January 1992 to April 1993. A pleuropneumoniae was isolated from 17(27.0%) of 63 growing pigs with respiratory signs and 21(6.4%) of 330 pneumonic lungs of slaughtered pigs. The seasonal isolation frequency of A pleuropneumoniae was higher in winter and spring than that in summer or fall. The biochemical and cultural properties of A pleluropneumoniae isolated from the pneumonic lungs of pigs were identical to those of the reference strains used. The isolates were highly susceptible to ampicillin, cephalothin, ceftiofur, ciprofloxacin(MIC : ${\leq}0.39{\mu}g/ml$) and moderately susceptible to amikacin, chloramphenicol, erythromycin, kanamycin, methicillin, penicillin-G, streptomycin(MIC : 0.78~25IU or ${\mu}g/ml$), respectively. Sulfadimethoxine, sulfamerazine, tylosine showed no response to the isolates(MIC : ${\geq}100{\mu}g/ml$). Among the 38 isolates, 21(55.3%) and 13(34.2%) were resistant to oxytetracycline aid lincomydn, respectively(MIC : ${\geq}50IU$ or ${\mu}g/ml$). The majority of 38 A pleuropneumoniae isolates were turned out as serotype 2(47.4%) or serotype 5(54.7%) and the remaining 3 isolates were evenly classified to serotype 7, 10 or 12. It was noted A pleuropneumonine serotype 5 isolates were more resistant to oxytetracycline than serotype 2 isolates.
정병열,Jung, Byeong-yeal 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.3
Haemophilus somnus, Mycoplasma bovis and Pasteurella multocida were responsible for respiratory diseases in bovine. Methods for identifying these bacteria had poor sensitivity and specificity. In this paper, PCR assays were applied for rapid identification of H. somnus, M. bovis, P. multocida B:2 and P. multocida capsular types. The specific PCR products were amplified from H. somnus, but not from other bacteria. Ten-fold diluted H. somnus were mixed with P. multocida and then the mixed cultures were inoculated on agar plates. After incubation, PCR was performed with harvest from agar plates and could detect as few as 3.4 CFU/ml of H. somnus. The primers MboF and MboR produced an amplification product unique to M. bovis and sensitivity of PCR was as low as 100 pg of DNA. Only serotype B:2 of P. multocida, the causal agent of haemorrhagic septicemia in bovine, was specifically amplified in PCR among the 16 reference serotypes. The multiplex capsular PCR typing for P. multocida was produced the P. multocida specific product as well as the capsular serogroup-specific product. The present PCR assays should be useful for the rapid identification of bacterial pathogens from bovine respiratory diseases.