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.
ABSTRACT. In total, 582 sera from 116 black goat herds were analyzed by a commercially available ELISA kit to monitor the seroprevalence of Mycobacterium avium subspecies paratuberculosis (Mpt) in Korean black goats (Capra hircus aegagrus). The mean number of goats sampled per herd was 5.11, 4.66, and 5.38 for the northern, central, and southern regions of Korea, respectively. The apparent regional prevalence of Mpt was estimated at 18.2-38.2% and 4.6-15.3% for herds and goats, respectively. The Mpt-positive goats were predominantly detected in the south (n=28), compared to either the northern (n=9) or central (n=11)
ABSTRACT. From 2002 to 2005, we collected 118 serum samples from 34 species belonging to 13 families of zoo animals in Korea and determined the prevalence of antibodies for 18 serovars of Leptospira spp. using the microscopic agglutination test (MAT). Twentynine (25%) of the serum samples tested were positive for one or more of the serovars. There were no significant differences in relation to genders: 23% and 26% of positives occurring in male and female animals, respectively (P>0.05). However, the seroprevalence for the Leptospira spp. was significantly higher (P<0.05) in herbivores (45%) than in either carnivores (17%) or omnivores (17%). Among the 5 serovars detected in this study, the most common was sejroe (n=27; 87% of all positive reactions). All positive reactions showed low titers (≤1:200) and the positives were most frequently detected in 1:25 (58%) and 1:50 (23%) serum dilutions. The highest antibody titer (1:200) was observed for the serovars sejroe (n=1) and bratislava (n=1). We conclude that the exposure of zoo animals to Leptospira spp. is relatively common in Korea and produces low MAT titers, with sejroe being the most commonly encountered serovar. KEY WORDS: leptospirosis, seroprevalence, zoo animals.
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