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Detection of infectious canine hepatitis virus by TaqMan real-time PCR method
Wang, H.Y.;Choi, J.Y.;Lee, M.J.;Park, J.H.;Cho, M.R.;Han, J.C.;Choi, K.S.;Chae, J.S.(Chonbuk National University, Jeonju, Republic of Korea)E-mail:jschae@chonbuk.ac.kr
The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples.
Afficher plus [+] Moins [-]Specific detection of Salmonella serogroup D1 by polymerase chain reaction(PCR) for sefA gene
1999
Jun, M.H. | Kim, T.J. | Chang, K.S. | Kang, K.I. | Kim, K.H. | Kim, H.S. | Shin, K.S. | Kim, C.J. (Chungnam National University, Taejon (Korea Republic). College of Veterinary Medicine) | Kim, K.S. (National Veterinary Research and Quarantine Service, Anyang (Korea Republic).) | Yoo, S.S. (Taejon City Institute of Health and Environment, Taejon (Korea Republic).)
Sal enteritidis thin fimbriae, SEF14, were found to be restricted to the predominantly poultry-associated members of the Salmonella serogroup D1 that are considered as the important pathogens in poultry industry. SefA together with sefB and sdfC encode the proteins involved in SEF14 biosynthesis. In order to develop the rapid and specific detection methods for Salmonella serogroup D1, a PCR technique for the am;lification of sefA gene was established, and its specificity and sensitivity were investigated with various microorganisms. The bacterial genomic DNA was extracted by colony-picking and rapid boiled-lysate technique. In comparison of SefI and SefII primers used in the PCR. SefI primer for sefA gene of 513bp showed higher specificity than that of SefII. The established PCR was s sensitive as to detect 1pg of Sal enteritidis DNA. When 73 strains in 28 genera including the reference strains and the field isolates of various Salmonella serotypes, Bacillus subtilis, Bordetella bronchiseptica, E coli, Listeria spp., Micrococcus luteus, Rhodococcus equi, Staphylococcus spp., Streptococcus spp., Vibrio parahemolyticus, Yersinia spp. were studied, the established PCR yielded specifically positive results with only Salmonella serogroup D1. The results suggested that the PCR for sefA gene could be a potential candidate among the specific detection methods for Salmonella serogroup D1.
Afficher plus [+] Moins [-]Identification of Hanwoo and Holstein meat using MGB probe based real-time PCR associated with single nucleotide polymorphism (SNP) in Melanocortin 1 receptor (MC1R) gene
2005
Park, S.D. (Chonnam National University, Gwangju, Republic of Korea) | Kim, T.J. (Chonnam National University, Gwangju, Republic of Korea) | Lee, J.I. (Chonnam National University, Gwangju, Republic of Korea), E-mail: jaeil@chonnam.ac.kr
The melanocortin 1 receptor (MC1R) plays an important role in regulation of melanin pigment synthesis within mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat color variations within several mammalian species including cattle. To develope a rapid and accurate method for the identification of Hanwoo meat, we performed a single nucleotide polymorphism (SNP) analysis in Melanocortin 1 receptor (MC1R) gene using TaqMan∨® MGB probe-based real-time PCR. Two specific probes (one for Hanwoo and the other for Holstein and Black angus) were designed.
Afficher plus [+] Moins [-]Sequence analysis and expression of groE gene encoding heat shock proteins of Brucella abortus isolates
2005
Kim, T.Y. (ChoongAng Vaccine Laboratory, Daejeon, Republic of Korea) | Kim, J.Y. (Chungnam National University, Daejeon, Republic of Korea) | Chang, K.S. (University of Kentucky, Lexington, KY, USA) | Han, H.R. (Seoul National University, Seoul, Republic of Korea) | Kim, M.C. (Chungnam National University, Daejeon, Republic of Korea) | Park, C.S. (Chungnam National University, Daejeon, Republic of Korea) | Jun, M.H. (Chungnam National University, Daejeon, Republic of Korea), E-mail: mhjun@cnu.ac.kr
GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains.
Afficher plus [+] Moins [-]Rapid differentiation of Hanwoo and Holstein meat using multiplex allele specific polymerase chain reaction protocols
2005
Koh, B.R.D. (Gwangju Metropolitan Health and Environment Research Institute, Gwangju, Republic of Korea), E-mail: barada@nvrqs.go.kr
Here I describe a multiplex allele specific PCR-based approach for the rapid detection between Hanwoo and Holstein meat associated with Melanocortin 1 receptor (MC1R) gene. Specific and universal oligonucleotide primers were used in combination to detect the presence of a single nucleotide polymorphism within the bovine MC1R DNA sequence. The presence of the bovine MC1R gene is indicated by the production of a single control PCR product, whilst positive samples generate an alternative smaller specific product over the same region. The mutations in MC1R104 codon revealed depending on the presence or absence of an indicative fragment amplified from the wild-type allele of this codon.
Afficher plus [+] Moins [-]Paternity test in dogs by microsatellite allele analysis
1999
Chae, Y.J. | Kim, D.K. | Kim, H.N. | Lee, M.H. | Hwang, W.S. | Lee, B.C. | Youn, H.Y. | Lee, H. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine)
Microsatellite allele analysis has been used for individual identification and paternity test. In the present study, the biological father of three puppies was determined by using microsatellite allele amplification analysis. The mother bitch of the litter was a poongsan dog. The three study dogs that could have inseminated the bitch, by being in the same residence, were a white Poosan dog, a mixed breed, and a white Jindo dog. DNA was obtained from all the relevant dogs by buccal swabbing. Four loci of tetranucleotide repeat microsatellite were PCR-amplified, and analyzed by polyacrylamicde gel electrophoresis and silver staining. The results of genotyping unambigously assigned the Poongsan dog as the biological father. There was no evidence of superfecundation. Therefore, the present study demonstrated the usefulness of microsatellite allele analysis as a simple, efficient method of paternity test in dogs.
Afficher plus [+] Moins [-]Analysis of genetic diversity for cattle parentage testing using microsatellite markers
Cho, G.J.;Yang, Y.J.(Korea Racing Association, Gwachon, Republic of Korea)E-mail:chogj@kra.co.kr | Lee, K.W.(Miryang National University, Miryang, Republic of Korea)
The objective of present study was to ascertain genetic diversity for cattle parentage testing. A total of 59 random cattle samples(29 Korean native cattle and 30 dairy cows) were genotyped by using 11 microsatellite loci(BM1824, BM2113, ETH10, ETH225, EH3, INRA23, SPS115, TGLA122, TGLA227, TGLA53, and TGLA126). This method consisted of multiplexing PCR procedure and showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer.
Afficher plus [+] Moins [-]Development of a multiplex-PCR for the rapid detection of Escherichia coli O157:H7 from raw beef
1998
Jung, S.C. | Jung, B.Y. | Cho, Y.S. | Kim, J.Y. (National Veterinary Research Institute, RDA, Anyang (Korea Republic). Department of Bacteriology) | Yoon, J.W. | Park, Y.H. (Seoul National University, Suwon (Korea Republic). Department of Veterinary Medicine)
Studies on hemoglobin type in Cheju native horse
1991
Hyun, H.S. | Kim, W.K. | Han, B.K. (Chonnam National Univ., Kwangju (Korea Republic). Coll. of Veterinary Medicine)
Restriction endonuclease analysis of canine parvovirus DNA isolated in Korea
1992
Park, J.H. | Song, J.Y. | Lee, J.B. | Hyun, B.H. | An, S.H. (Rural Development Administration, Anyang (Korea Republic). Veterinary Research Inst.) | Jun, M.H. (Chungnam Nat'l Univ., Taejon (Korea Republic). Coll. of Veterinary Medicine)