• Korean Journal of Microbiology
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• v.39 no.1
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• pp.40-44
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• 2003

Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. In order to develop a DNA marker specific for Bacillus anthracis and to discriminate this species from Bacillus cereus group, we applied the randomly amplified polymorphic DNA (RAPD)-PCR technique to a collection of 29 strains of the genus Bacillus, including 22 species of the B. cereus group. A 709-bp RAPD marker (KHT5) specific for B. anthracis was obtained from B. anthracis BAK. The PCR product of internal primer set from the KHT5 fragment distinguished B. anthracis from the other species of the B. cereus group.

• Korean Journal of Plant Resources
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• v.16 no.1
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• pp.40-48
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• 2003

Cylindrocarpon destructans is the major pathogen inducing the root rot disease in ginseng. Up to now, there is no reliable and convenient method to analyze the spore density or population of this pathogen in ginseng-growing soil or any contaminated farmlands. Therefore, it will be very valuable to develop a new and reliable method in detecting the spore of this pathogen. In this study, a molecular biological technique using two step nested PCR method, was developed. Two universal ITS primers, ITS5F and ITS4R were used in the first round of PCR to amplify a fragment of ITS region from the genomic DNA of C. destructans. The specific prmers Nest 1 and Nest 2 were designed and used in the second round of PCR to amplify a inner fragment from the first round PCR product of C. destructans. C. destructans spore, only soil samples from the diseased ginseng farm produced the positive bands, suggesting its usefulness in detecting the C. destructans spores in soil samples. Thus it is recommended to first extract the whole genomic DNA from soil samples and use it for the PCR reaction, thereby eliminating the inhibitory activity of soil components.

• Horticultural Science & Technology
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• v.28 no.4
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• pp.672-677
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• 2010

A duplex PCR method was developed to detect a transformation vector pSPB130 used in the development of a genetically modified (GM) rose plant. To detect a GM rose plant, the anthocyanin synthase ($ANS$) was used as an endogenous reference gene of rose in PCR detection. The primer pair RHANS-KF/KR producing 107 bp amplicon was used to amplify the $ANS$ gene and no amplified product was observed in any of the 9 different plants used as a template. The primer pair GMRH-KF/KR was designed to amplify the junction sequence between 35S promoter and flavonoid 3',5'-hydroxylase ($F3^{\prime}5^{\prime}H$) gene in pSPB130. The detection limit of the duplex PCR method is approximately 0.5%. This result indicates that this duplex PCR method could be useful for monitoring unauthorized GM rose in Korea.

• Journal of the Korean Chemical Society
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• v.47 no.2
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• pp.127-132
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• 2003

We have developed a direct polymerase chain reaction (PCR) and electrophoresis method for the diagnosis of bovine theileriosis from whole blood DNA analysis without DNA extraction. The technique empolyed a FoLT (formamide, low temp.) technique and was utilized in the diagnosis of bovine theileriosis. Formamide solubilize the blood cells, and the lowered incubation temperatures reduced protein coagulation. 100-200 nL of whole blood and PCR reagents were introduced directly into a PCR tube. After the amplification, the PCR product (816-bp DNA) was introduced into the electrophoresis system. The results of this analysis were consist with those obtained using purified DNA.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.3
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• pp.275-282
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• 1993

The objective of this study was to clone a partial fragment of $\alpha$-amylase from Korean maize. We designed and synthesized an oligonucleotide probe and two kinds of PCR primers based on cDNA conserved region of $\alpha$-amylase sequences from other plants. Total RNA from 3-day-old maize seedling was used as template for 1st strand cDNA synthesis and RNA-DNA hybrid was used as template for polymerase chain reaction(PCR). The product of PCR was about 0.5 kb long and inserted into pUC19. We named this recombinant plasmid as pZM$\alpha$'. The cloned fragment was certified by Southern blot analysis using labeled synthetic oligonucleotide as probe.

• Korean Journal of Veterinary Service
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• v.24 no.4
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• pp.335-341
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• 2001

Swine dysentery caused by Brachyspira hyodysenteriae, an anaerobic, beta-hemolytic spirochete, is a severe mucohemorrhahic diarrheal disease that primarily affects pigs during the growing and finishing period. The current standard laboratory procedure to culture and identify B hyodysenteriae takes 3 to 7 days. This report present a rapid PCR for detection B hyodysenteriae in a single reaction using DNA from swine intestinal samples. The PCR produced a specific 421bp PCR product with template DNA purified from B hyodysenteriae, and the accuracy for detection of B hyodysenteriae by PCR results compared with those of conventional method was 100% in intestinal specimens. Nonspecific bands were not detected with B innocens, a nonpathogenic common inhabitant spirochete, including other enteric bacterial organisms. This procedure could detect as little as 50 pg of template DNA for B hyodysenteriae.

• The Plant Pathology Journal
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• v.30 no.2
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• pp.178-182
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• 2014

Among imported plants, seeds are the items that have many latent pathogens and are difficult to inspect. Also, they are the import and export items whose market is expected to expand. The biggest problem with seeds is viruses. Prune dwarf virus (PDV) is the virus that is commonly inspected in Prunus cerasifera, P. persica, P. armeniaca, P. mandshurica, P. cerasus, P. avium or P. serotina seeds. In this study, two RT-PCR primer sets, which can promptly and specifically diagnose plant quarantine seed-transmitted PDV, were developed; and nested PCR primers, where products amplify 739 and 673 nucleotides (nt), and an nested PCR-product, 305 nt, can be obtained as these products are amplified again, were developed. Also, a modified-positive control plasmid was developed, where the restriction enzyme XhoI, which can identify the contamination of samples from the control, was inserted. The method developed in this study has detected PDV in 18 cases since 2007, and is expected to continuously contribute to the plant quarantine in Korea.

• Korean Journal of Veterinary Service
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• v.29 no.1
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• pp.1-8
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• 2006

Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.

• Korean Journal of Veterinary Service
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• v.26 no.2
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• pp.105-111
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• 2003

From February 2000 to December 2001, A total of 1,785 samples was taken from beef and pork carcasses in Seoul. Seven(0.69%) Listeria spp. were isolated from the 1,014 of beef carcasses, and five(0.65%) were isolated from the 771 of pork carcasses. The isolates were identified L monocytogenes by API listeria, and VIDAS LMO kit, serological test and PCR assay were preformed. A total 12 strains of L monocytogenes were isolated form samples tested and all of the strains were classified into serotype 1. PCR primers are selected to amplify a 520-base pair(bp) DNA fragment from the listeolysin O gene(hlyA) of Listeria monocytogenes. A 520-bp product was detected in PCR with DNA from L monocytogenes, but not from the other Listeria species tested.

• Korean Journal of Veterinary Service
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• v.37 no.4
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• pp.247-252
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• 2014

The economic impact of swine mycoplasma infection is high. An accurate diagnosis is often difficult and time consuming. We report the development and validation of an effective multiplex polymerase chain reaction (PCR) assay that detects Mycoplasma (M.) hyopneumoniae and M. hyorhinis. The multi detection of M. hyopneumoniae and M. hyorhinis primer set were employed to detect mycoplasma species and typing of the species was performed on the basis of sequence analysis of the PCR product. The target nucleic acid fragments were specifically amplified by M. hyopneumoniae and M. hyorhinis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were used to evaluate the specificity of the multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. hyopneumoniae and M. hyorhinis.