• Journal of the East Asian Society of Dietary Life
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• v.18 no.5
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• pp.753-759
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• 2008

The principal objective of this study was to develop the optimum oligonucleotide primers for the simple detection of Campylobacter in food samples. In order to achieve this goal, a variety of oligonucleotide primers were designed via the modification of 16S rDNA, ceuE and mapA sequences of Campylobacter. Through the subsequent analysis of the specificity and sensitivity of primers, two types of oligonucleotide primers, CB4 and CJ1, were selected for Campylobacter genus-specific and C. jejuni species-specific primers, respectively. The detection limit was found to be $10^0{\sim}10^1$ cells per reaction with the prepared cell suspension, however, the sensitivity in the meat samples was less, at $10^1{\sim}10^2$. We suggested that PCR inhibitors such as hemoglobin or immunoglobulin in pork or beef influenced.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2001.12a
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• pp.90-90
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• 2001

• Korean Journal of Veterinary Service
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• v.30 no.4
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• pp.527-532
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• 2007

In present study, we described the reliability of the dual priming oligonucleotide (DPO) multiplex polymerase chain reaction (PCR) for the detection of Mycobacterium tuberculosis complex (MTC) and non-Mycobacterium tuberculosis (NMT) in blood samples of the Korea native cattle, Hanwoo. Among 340 samples 22 (6.5%) were positive in using DPO multiplex PCR, 21 (6.2%) were positive in PCR. The relative agreement between 2 tests was 99.7%, and the agreement quotient (kappa), was 0.95 (excellent). In these results, we demonstrated the successful application of DPO multiplex PCR for the diagnosis of bovine tuberculosis in Hanwoo. Multiplex PCR, using DPO primers, can be useful for the simple diagnosis of bovine tuberculosis in bovine blood samples.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.2
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• pp.295-300
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• 2009

The specificity and sensitivity of oligonucleotide primers were examined for the rapid detection of Salmonella in meat samples. The oligonucleotide primers used in this study were designed with the modification of mdh and invA sequence in the chromosome of Salmonella Typhimurium. Through the subsequent analysis of the specificity and sensitivity of the primers, two types of oligonucleotide primers, SLM1 and SLT4 were selected for the detection of Salmonella genus specific and S. Typhimurium species specific, respectively. The lowest detection limit of each primer was represented as 1 cell per reaction when reacted with a prepared DNA solution. The detection efficiency of the two primers was analysed with beef and pork samples intentionally contaminated with a mixture of Salmonella culture, and three preparation methods -, namely direct reaction after extraction, enrichment after extraction, and DNA extraction after enrichment for PCR reaction, - were also compared. No differences were found in the results according to meat sources and preparation methods.

• Journal of Life Science
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• v.8 no.3
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• pp.285-293
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• 1998

We have cloned an internal fragment of the putative transoisase gene of MLE in the silkworm, Bombyx mori, using PCR method with degenerative oligonucleotide primers designed to represent regions of amino acids encoding transposase. The resulting PCR clone, designed as BmoMAR, cords a partial ORF(152 a.a.) of MLE in which interrupted by five stop codons, and the sequence of its deduced amino acids showed 37% homology with Mos1, an active mariner, from Drosophila mauritiana. Furthermore, the BmoMAR exhibits nucleotide and amino acid homology with 59% and 37% from Apis mellifera and D. mauritiana 7.9 clone, respectively. This result strongly that a MLE is present in the genome of B. mori.

• Biomedical Science Letters
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• v.4 no.1
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• pp.43-56
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• 1998

A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I.II), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) O157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I.II), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5$\times$10$^{6}$ of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.6
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• pp.761-769
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• 1996

The random amplified polymorphic DNA (RAPD) technique was used to characterize 18 reference strains of microalgae, mostly Chlorella species, collected from various localities around Korea peninsular. Eighteen strains consist of four genera of the family marine Chlorella from 12 samples, two genera of fresh water Chlorella from three samples, and three genera on Nannochloris. Twenty 10-mer anonymous primers were screened for amplification of genomic DNA extracted from samples using the CTAB extraction method. Nineteen of these oligonucleotide primers were positive or band producing. Three of 20 random primers (OPA 10, OPA 12, and OPA 18) resulted in both clear band and a high degree of reproducibility and showed some potential to be used to discriminate individual samples of both genetically hetero-and homogeneous populations, in determining phylogenetic relationships between species within a genus and developing individual fingerprints for each samples.

• Korean Journal of Oriental Medicine
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• v.7 no.1
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• pp.145-152
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• 2001

Dried parts of the herb medicines are difficult to distinguish morphologically. Heracleum moellendorffii cordata has often been sold instead of Aralia cordata in herbal medicine markets. Therefore, this study was conducted to develop the key for discrimination between them using the RAPD analysis and morphological characteristics. Thirty decarmer oligonucleotide primers were screened for the RAPD analysis, and four primers generated distinct RAPD markers specific to Aralia cordata, Angelica pubescens maxim f. biserrata, and Heracleum moellendorffii. The specific RAPD patterns generated by the selected primers were reproducible from dried materials. In comparison of morphological characteristics, Aralia cordata seems to be entirely developed in xylem fiber, but not developed in pith.

• Mycobiology
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• v.30 no.4
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• pp.202-207
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• 2002

This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

• The Plant Pathology Journal
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• v.20 no.2
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• pp.147-154
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• 2004

A simple and reliable procedure for RT-PCR detection of Apple stem pitting virus (ASPV), Cherry rasp leaf virus (CRLV), and Cherry necrotic rusty mottle virus (CNRMV) was developed. Two virus specific primer sets for each virus were found to specifically detect each virus among fourteen sets of designed oligonucleotide primers. Total RNAs extracted from healthy and from ASPV-,CRLV- and CNRMV-infected plant tissues were used to synthesize cDNA using oligo dT primer and then amplified by virus-specific primers for each virus. Each primer specifically amplified DNA fragments of 578 bp and 306 bp products for ASPV (prAS CP-C and prAS CP-N primers, respectively); 697 bp and 429 bp products for CRLV (prCR4 and prCR5-JQ3D3 primers, respectively); and 370 bp and 257 bp products for CNRMV (prCN4 and prCN6-NEG 1 primers, respec-tively) by RT-PCR. DNA sequencing of amplified DNA fragments confirmed the nature of each amplified DNA. Altogether, these results suggest that these virus specific primer sets can specifically amplify viral sequences in infected tissues and thus indicate that they can be used for specific detection of each virus.