• Korean Journal Plant Pathology
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• v.12 no.2
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• pp.191-196
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• 1996

위축, 황화, 총생 증상 등 전형적인 병징을 나타내는 phytoplasma에 감염된 식물에서 phytoplasma만을 특이적으로 검출하기 위하여 polymerase chain reaction(PCR) 방법을 이용하였다. Phytoplasma의 16S rRNA gence의 DNA 단편을 증폭하기 위하여 1.4 kb primers (forward, 5` -GTTGATCCTGGCTCAGGATT-3` 와 reverse, 5` -AACCCCGAGAACGTATTCACC -3`)를 사용하여 증폭한 결과, phytoplasma에 이병된 오동나무, 라일락 및 미역취에서는 약 1.4 kbp의 위치에서 특이 band가 검출되었으나 control로 사용한 건전주에서는 어떠한 band 검출되지 않았다. 위의 결과를 재확인 하기 위하여 약 0.5 kb의 primers(forward, 5` -ACGAAAGCGTGGGGAGCAAA-3` 와 reverse, 5` -GAAGTCGAGTTGCAGACTTC-3`)를 사용하여 증폭한 결과, 0.5 kb의 위치에서 특이 band가 검출되었으나 control로 사용한 건전주에서는 어떠한 band도 검출되지 않았다. Phytoplasma에 이병된 식물의 PCR 반응산물을 제한효소인 AluI으로 처리하 sruf과, 오동나무와 라일락에서는 동일한 band pattern을 나타내어 서로 유연관계가 가까운 phytoplasma인 것으로 생각되며, 미역취에서는 이들과는 다른 band pattern을 나타내어 오동나무와 라일락의 phytoplasma와는 유연관계가 먼 것으로 추측된다.

• Journal of Environmental Health Sciences
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• v.28 no.2
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• pp.193-202
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• 2002

The polymerase chain reaction(PCR) of target lacZ and uidA genes were used to detect total coliform and Escherichia coli for determining water quality, respectively. Of 109 spring waters, coliform were detected from 38 spring waters by lacZ PCR method but 21 spring waters by culture method accepted by the Ministry of Environment for water quality monitoring. The lacz PCR method gave the results statistically equivalent to those of the culture method(kappa=0.62, McNemar=17.00). The uidA PCR method gave the same results to those of the culture method. The sensitivity and specificity of coliform and E. coli by PCR method were 100% and 80.7%, respectively. Therefore, PCR can be used for the rapid identification of Escherichia coli and coliform in potable water using uidA and lacZ.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.90-95
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• 1999

A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum(M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.

• Korean Journal of Microbiology
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• v.40 no.3
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• pp.254-256
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• 2004

Polymerase chain reaction (PCR) is a powerful tool for precisely amplifying selected DNA sequences that have had a broad impact on genomic studies. When examining human $\alpha$- and $\beta$- tryptase genes which have 95% DNA homology, inconsistent PCR amplification of genomic sequences hampered our progress. This study suggests that long PCR technique on the original DNA digested with restriction enzymes improves both efficiency and sensitivity of PCR. These improved results seem to derived from the effective denaturation of the original genomic DNA template or reduction of formation of secondary structures that block either primer annealing or extension in PCR. Elimination of homo- or hetero-duplex products derived from highly homologous genes provides an additional advantage in this study. This communication describes how the use of restriction enzymes improved these efficiencies, and also facilitated studies of highly homologous genes including tryptase genes.

• Journal of fish pathology
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• v.21 no.3
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• pp.181-188
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• 2008

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used to detect and identify four fish viruses, fish iridovirus, viral hemorrhagic septicaemia virus (VHSV), viral nervous necrosis virus (VNNV), hirame rhabdovirus (HRV). Four viruses were detected by PCR with each specific primers. Identification of iridovirus was achieved by digesting the PCR amplified fragment with a restriction enzyme ApaⅠ. It was possible to distinguish positive from false positive PCR amplicons of VHSV by RFLP of PstⅠ or HindⅢ restriction enzymes. VNNV was identified using RFLP of BamHⅠrestriction enzyme and HRV was identified by XbaⅠ restriction enzyme. This approach can be used for more rapid, simple and specific diagnosis of fish viral diseases.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.6
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• pp.740-744
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• 2014

In this study, the ${\beta}$-lactamase (VPA0477) gene was used as a new target for the PCR-based detection of Vibrio parahaemolyticus. Primers specific for the ${\beta}$-lactamase (VPA0477) gene of V. parahaemolyticus, were designed and incorporated into a PCR-based assay. The assay was able to specifically detect all of the 191 V. parahaemolyticus strains tested, but did not result in amplification of 39 other Vibrio spp. and non-Vibrio spp. strains tested. The detection limit of the assay was 10 CFU of V. parahaemolyticus RIMD2210633 from pure culture broth. The ${\beta}$-lactamase (VPA0477) gene-based assay developed in this study was sensitive and specific, and has great potential for the accurate detection and identification of V. parahaemolyticus in seawater or seafood samples.

• Korean Journal of Veterinary Service
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• v.23 no.3
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• pp.227-233
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• 2000

Salmonella enteritidis is the most prevalent etiologic agents of foodborne acute gastroenteritis. Direct isolation and identification of S enteritidis are time consuming work and not so highly sensitive. This study was conducted to develop for the specific detection of S enteritidis using polymerase chain reaction(PCR). PCR primers were selected to amplify a 351-base pair(bp) DNA fragment from the salmonella plasmid virulence A(spv A) gene of S enteritidis. With the primers, 351 bp DNA products were amplified from S enteritidis but not from other B, D, Cl serogroup Salmonella spp. It was sensitive to detect up to 40 pg of template DNA by agarose gel electrophoresis. This PCR assay is very rapid and specific method and less time consuming than the standard bacteriological methods.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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• pp.213-220
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• 2003

Polymerase chain reaction (PCR) is one of the most widely used analytical tool and is an important module that would benefit from being miniaturized and integrated onto diagnostic or analytical chips. There are potentially two different approaches for the miniaturization of the PCR module: chamber-type and flow-type micro-PCR. These miniaturized PCRs have distinct characteristics and advantages. In this article, we review the necessity of micro-PCR, the materials for the chip fabrication, the surface modification, and characteristics of the two types of micro-PCR. The motivation underlying the development of micro-PCR, the advantages and disadvantages of the various materials used in fabrication and the surface modification methods will be discussed. And finally, the precise features of the two different types of micro-PCR will be compared.

• Korean Journal of Clinical Laboratory Science
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• v.37 no.3
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• pp.139-142
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• 2005

Most expressed HLA(human leukocyte antigen) loci exhibit a remarkable degree of allelic polymorphism, which is derived from sequenceing differences predominantly localized to discrete hypervariable regions of the amino-terminal domain of the molecule. In this study, the HLA-DRB1 genotypes were determined in twenty students using the PCR-SSP (polymerase chain reaction-sequence specific primer) technique. Two specific primer pairs in assigning the DRB1 gene were used. The results of PCR-SSP, the $HLA-DRB1^{\ast}0101$ primer detected nine and $HLA-DRB1^{\ast}1501$ primer detected three people. This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-DRB1 genotypes. Moreover, these genotype frequency results of the HLA DRB1 gene could be useful for database study before being applied to individual identification and transplantation immunity.

• Journal of fish pathology
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• v.12 no.1
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• pp.66-69
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• 1999

Occurrences of red sea bream iridovirus disease (RSIVD) in cultured marine fishes were investigated. The infection was detected by the polymerase chain reaction (PCR) used to amplify the red sea bream iridovirus (RSIV). The RSIV infection was widely distributed in fish culture farm around the south coastal area of the Korean peninsula.