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Su Hwa Lee Byeong Yeal Jung Nabin Rayamahji Hee Soo Lee Woo Jin Jeon Kang Seuk Choi Chang Hee Kweon Han Sang Yoo 《Journal of veterinary science (Suw?n-si, Korea)》2009,10(1):43-51
Salmonella (S.) Typhimurium and S. Enteritidis are the major causative agents of food-borne illnesses worldwide. Currently, a rapid detection system using multiplex real-time polymerase chain reaction (PCR) has been applied for other food-borne pathogens such as Escherichia coli, Staphylococcus aureus and Streptococcus spp. A multiplex real-time PCR was developed for the simultaneous detection of Salmonella spp., especially S. Typhimurium and S. Enteritidis, in beef and pork. For the specific and sensitive multiplex real-time PCR, three representative primers and probes were designed based on sequence data from Genbank. Among the three DNA extraction methods (boiling, alkaline lysis, and QIAamp DNA Mini Kit), the QIAamp DNA Mini Kit was the most sensitive in this study. The optimized multiplex real-time PCR was applied to artificially inoculated beef or pork. The detection sensitivity of the multiplex real-time PCR was increased. The specificity of the multiplex real-time PCR assay, using 128 pure-cultured bacteria including 110 Salmonella isolates and 18 non-Salmonella isolates, was 100%, 100% and 99.1% for Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. The sensitivity was 100%, 100% and 91.7% for Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. The multiplex real-time PCR assay developed in this study could detect up to 0.54 ± 0.09 and 0.65 ± 0.07 log10 CFU/ml for S. Typhimurium and S. Enteritidis for beef, 1.45 ± 0.21 and 1.65 ± 0.07 log10 CFU/ml for S. Typhimurium and S. Enteritidis for pork, respectively, with all conditions optimized. Our results indicated that the multiplex real-time PCR assay developed in this study could sensitively detect Salmonella spp. and specifically differentiate S. Typhimurium from S. Enteritidis in meats. 相似文献
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K Nakajima C H Kweon S H An T Shibayama N Wakamiya S Kato K Hirai 《Avian diseases》1990,34(2):479-484
Among the 33 monoclonal antibodies (MAbs) against pseudorabies virus (PRV) examined, three MAbs (24-17, 74-26, and 8) were found to react with cells infected with Marek's disease virus (MDV)-related viruses by immunofluorescence test. Two of the MAbs (24-17 and 74-26) reacted with the nuclei of cells infected with MDV serotype 1 (MDV1), MDV serotype 2 (MDV2), and herpesvirus of turkeys (HVT), whereas MAb 8 reacted with the cytoplasm of MDV2- and HVT-infected cells. However, none of the MAbs against MDV1, MDV2, and HVT that were examined reacted with PRV-infected cells. None of these three MAbs against PRV reactive with MDV-related viruses cross-reacted with the cells infected with other herpesviruses, such as herpes simplex virus type 1, herpes simplex virus type 2, varicella zoster virus, Epstein-Barr virus, or human herpesvirus 6. Southern-blot hybridization under stringent or less-stringent conditions showed that no significant DNA homology was detected between PRV DNA and MDV DNA. 相似文献
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Detection of porcine epidemic diarrhea virus by immunohistochemistry with recombinant antibody produced in phages 总被引：1，自引：0，他引：1
Lee HM Lee BJ Tae JH Kweon CH Lee YS Park JH 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(3):333-337
Several diagnostic methods including immunofluorescence, enzyme-linked immunosorbent assay, polymerase chain reaction and immunohistochemistry have been developed for the detection of porcine epidemic diarrhea virus (PEDV). An immunohistochemical method using a new recombinant antibody produced by a phage antibody system (PAS16) kit was investigated and compared with that using a monoclonal antibody for PEDV detection in PEDV-infected piglets. In both the immunohistochemical methods, PEDV antigens were detected in the cytoplasm of villous enterocytes and in the macrophages infiltrated in the lamina propria at 18 to 110 hr post inoculation. The positive signals with the recombinant PAS16 antibody were similar to those with the monoclonal antibody. This result suggests that the recombinant PAS16 antibody can be applicable for the rapid immunohistochemical diagnosis of PEDV infection. 相似文献
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Ji-Hey Lim Chang-Su Jung Ye-Eun Byeon Wan Hee Kim Jung-Hee Yoon Kyung-Sun Kang Oh-kyeong Kweon 《Journal of veterinary science (Suw?n-si, Korea)》2007,8(1):89-94
A model that provides reproducible, submaximal yet sufficient spinal cord injury is needed to allow experiments leading to development of therapeutic techniques and prediction of clinical outcome to be conducted. This study describes an experimental model for spinal cord injury that uses three different volumes of balloon inflation and durations of compression to create a controlled gradation outcome in adult dogs. Twenty-seven mongrel dogs were used for this study. A 3-french embolectomy catheter was inserted into the epidural space through a left hemilaminectomy hole at the L4 vertebral arch. Balloons were then inflated with 50, 100, or 150 µl of a contrast agent at the L1 level for 6, 12, or 24 h and spinal canal occlusion (SCO) measured using computed tomography. Olby score was used to evaluate the extent of spinal cord injury and a histopathologic examination was conducted 1 week after surgery. The SCO of the 50, 100, and 150 µl inflations was 22-46%, 51-70%, and 75-89%, respectively (p < 0.05). Olby scores were diminished significantly by a combination of the level of SCO and duration of inflation in all groups. Olby scores in the groups of 150 µl-12 h, 150 µl-24 h, and 100 µl-24 h were 0.5, 0, and 1.7, respectively. Based on these results, a SCO > 50% for 24 h, and > 75% for 12 h induces paraplegia up to a week after spinal cord injury. 相似文献
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Park W Kim WH Lee CH Kim DY Choi JH Huh JW Sung HM Kim IS Kweon OK 《Journal of veterinary medicine. A, Physiology, pathology, clinical medicine》2002,49(7):385-389
To control intra-operative haemorrhage, fibrin glues are preferred by many surgeons because of their biological advantages and convenience of application. Manufacturers have developed a few kinds of fibrin glues with a little difference in their composition. This study was to compare the effectiveness of two commercially available fibrin glues; Greenplast (Green Cross P. D. Company, Yongin, Korea) and Tisseel (Baxter-Immuno AG, Vienna, Austria). They were applied experimentally to several kinds of surgery in dogs - renal vessel anastomosis, partial splenectomy, intestinal anastomosis and incision skin wound - and evaluated for their haemostatic and adhesive effects. When the two glues were applied in renal vessel anastomosis, the amount of haemorrhage in artery and vein decreased significantly. They also decreased the haemorrhage in partial splenectomy. At 10 min after application of the glues to an incision skin wound, the tensile strengths developed were significantly higher than that of control. The present study indicates that two-component fibrin glues have a haemostatic effect as a mechanical barrier in renal vessel anastomosis and an adhesive effect in the early stage of incision skin wound closure, and the two glues have similar effects with no complications. 相似文献
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Bovine herpes virus expressing envelope protein (E2) of bovine viral diarrhea virus as a vaccine candidate. 总被引：5，自引：0，他引：5
C H Kweon S W Kang E J Choi Y B Kang 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1999,61(4):395-401
The gene encoding the envelope protein (E2) of bovine viral diarrhea virus (BVDV) was expressed under the thymidine kinase (TK) promoter of Korean bovine herpesvirus 1 (BHV-1) isolate. Thymidine kinase negative (TK-) BHV-1 recombinants expressing E2 of BVDV were constructed and the expression of E2 was identified by immunofluorescence and Western blotting. Compared to wild type BHV-1, the recombinant BHV-1 had a delayed cytopathogenic effect in cells. The immunogenicity of the recombinant BHV-1 was examined in guinea pigs and cattle. Although an increase in body temperature was detected for a few days, the inoculated cattle returned to normal temperature with the development of neutralizing antibodies to BVDV. 相似文献