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Su Hwa Lee,정병열,Nabin Rayamahji,Hee Soo Lee,Woo Jin Jeon,Kang Seuk Choi,권창희,유한상 대한수의학회 2009 Journal of Veterinary Science Vol.10 No.1
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.
Kang, Mi Lan,Shin, Seung Won,Rayamahji, Nabin,Seo, Yeon Soo,Lee, Su In,Lee, Won Hyung,Yoo, Han Sang The Korean Society of Veterinary Science 2008 大韓獸醫學會誌 Vol.48 No.1
We have investigated efficiency of a recombinant subunit Pasteurella multocida toxin (PMT) that was mixed with a vaccine consisted of inactivated whole cells of Bordetella bronchiseptica, P. multocida (types A and D). For verification of the efficacy of the vaccine, all experimental pigs (suckling piglets, sow and gilts) in the three farms were vaccinated. Antibody titers against B. bronchiseptica and P. multocida type A of the vaccinated pigs by microplate agglutination were significantly higher than those of the control pigs (p < 0.05). Similar patterns were observed in the analysis of anti- PMT neutralizing antibody by serum neutralizing method using Vero cell (p < 0.05). Anti- P. multocida type D antibody titer of the vaccinated sows and gilts by ELISA showed significant differences with those of the non-vaccinated pigs (p < 0.05). Although antibody titers increased, it was unable to find out the difference in the clinical signs between the vaccinated and non-vaccinated pigs. However, the increase in body weight of the vaccinated piglets was observed in comparison with the non-vaccinated piglets on a farm. At slaughtering of the pigs, pathological lesions in the turbinate bones of the vaccinated pigs were significantly lower than those of the non-vaccinated pigs (p < 0.001). These results suggested that efficacy of the vaccine in pigs demonstrated to protect against atrophic rhinitis in Korea.
Seo, Yeon-Soo,Lee, Deog Yong,Rayamahji, Nabin,Kang, Mi Lan,Yoo, Han Sang Elsevier 2008 Research in veterinary science Vol.85 No.3
<P><B>Abstract</B></P><P>Biological monitoring is performed to detect and analyze microorganisms that have continuously made an effort to survive in the environment. Of such microorganisms, <I>Staphylococcus</I> spp. is considered a common cause of nosocomial and environmental infections., Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs) are required for the adhesion and biofilm formation of Staphylococci. Thirty-six and thirty-five Staphylococci isolated from animals and air, respectively, were analyzed. Biofilm formation and ten MSCRAMM genes were investigated using Congo red agar, tissue culture plate methods, and PCR. Airborne isolates were shown to have higher adherence and stronger biofilm formation than those from animals. The prevalence of MSCRAMM genes from air isolates was also higher than those from animals. Of the genes, <I>eno</I> was mainly associated with biofilm formation in both animals and airborne isolates (<I>P</I><0.05). Moreover, the rate of airborne isolates harboring the <I>eno</I> gene was higher than in animal isolates. These results indicated that analysis of MSCRAMM genes with a phenotypic assay might be a helpful bacterial control system for the environment.</P>