• 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.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.771-776
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• 1998

Synthetic oligonucleotide primers of 20 and 21 bases, respectively, were used in the polymerase chain reaction (PCR) to amplify a sequence of the mrp gene, which encodes the muramidase released protein of Streptococcus suis. Amplification was not recorded when 5 other streptococcal species were tested or when 9 different nonstreptococcal species were tested. A DNA fragment of 517bp was amplified from the genomic DNA of S suis. The lower detection limit was 100pg of the genomic DNA. The primers recognized 34 serotypes of S suis reference strains and 9 isolates from pneumonic lung, brain, nasal discharge, tonsil. This results suggest that the amplification of the mrp gene by PCR method is potential for the identification of S suis isolates.

• Fisheries and Aquatic Sciences
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• v.25 no.7
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• pp.403-408
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• 2022

In April and June 2014, mortalities of goldfish occurred at Korea. The principal signs included pale gills, severely enlarged and softened spleen and kidney and red liver. Moreover, the histopathological characteristics observed mainly in hematopoietic tissues of kidney, gill lamellae and intestine. The predominant histopathological changes were severe necrosis of hematopoietic tissue. In addition, nucleus exhibiting peripherally displaced chromatin were particularly abundant in the kidney of affected fish. The histological examination led to hypothesize the implication of a virus in the mortality. The hematopoietic necrosis herpesvirus (Cyprinid herpesvirus 2, CyHV-2) DNA polymerase gene successfully detected in DNA extracted from kidney and spleen of affected fish using PCR assay and showed 100% identity with already deposited CyHV-2 DNA polymerase gene in NCBI. Artificial infection trials using affected tissues filtrates gave cumulative mortalities of 30% for virus injected goldfish. In the present study, CyHV-2 was detected and identified as the causative pathogen of the epizootic in goldfish in Korea.

• Fisheries and Aquatic Sciences
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• v.26 no.11
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• pp.678-688
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• 2023

Flatfish are one of the largest families in the order Pleuronectiformes and are economically important edible marine fish species. However, they have similar morphological characteristics leading to challenges in classifying correctly, which may result in mislabeling and illegal sales, such as fraudulent labeling of processed food. Therefore, accurate identification is important to ensure the quality and safety of domestic markets in Korea. Species-specific primers were prepared from the mainly consumed eleven species of the order Pleuronectiformes. To rapidly identify the 11 flatfish species, a highly efficient, rapid, multiplex polymerase chain reaction (PCR) with species-specific primers was developed. Species-specific primer sets were designed for the mitochondrial DNA cytochrome c oxidase subunit I gene. Species-specific multiplex PCR (MSS-PCR) either specifically amplified a PCR product of a unique size or failed. This MSS-PCR analysis is easy to perform and yields reliable results in less time than the previous Sanger sequencing methods. This technique could be a powerful tool for the identification of the 11 species b the family Pleuronectidae and can contribute to the prevention of falsified labeling and protection of consumer rights.

• Journal of Oral Medicine and Pain
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• v.20 no.1
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• pp.229-246
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• 1995

The deoxyribonucleic acid(DNA) was isolated from the pulp, dentin and enamel of the 4 fresh teeth and the 7 teeth left in room temperature for 10 years. Then it was examed to find out the usefulness for forensic dental medicine. Samples of the tooth-derived DNA amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologous amelogenin gene and D1S80 locus detection. The following results have been achieved. DNA extraction was possible in pulp and dentin of the fresh teeth, so it could be applicatable to detection of X-Y homologous amelogenin gene for sex determination, amplification of D1S80 locus by PCR. Sex determination was possible in pulp and dentin of the teeth left at room temperature for 10 years. Also, possible the detection fo AMP-FLPs to increase PCR cycling up to 40. DNA was isolated from all pulp of the fresh teeth and the teeth left in room temperature for 10 years, and also isolated from the dentin of the fresh teeth, partially isolated(3/7) from the dentin of the teeth left in room temperature for 10 years, but DNA was not isolated from enamel. From the above investigation, DNA extraction, sex determination, amplification of D1S80 locus were successfully accomplished even though the teeth were left for 10 years at room temperature. Therefore, teeth, especially pulp, are highly reliable and applicable as molecular biological samples for individual identification.

• Food Science and Biotechnology
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• v.16 no.1
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• pp.67-70
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• 2007

Mackerel (Scomber japonicus) often causes severe allergic reactions in sensitive people. Food containing undeclared mackerel may pose a risk to such people. The major allergenic protein in fish such as mackerel, codfish, and Alaska pollack has been found to be parvalbumin. In this study, we developed a polymerase chain reaction (PCR) method to detect mackerel DNA using primers corresponding to the parvalbumin gene. We cloned and sequenced 1.5 kb of parvalbumin gene by PCR using mackerel genomic DNA as a template. Nucleotide sequence analysis of genomic parvalbumin gene, composed of 4 exons and 3 introns, allowed the selection of two pairs of oligonucleotide primers specific for mackerel. These primers successfully enabled PCR amplification of specific regions of genomic parvalbumin DNA from mackerel, but no amplification from 8 other fish samples, surimi, and 6 boiled fish pastes. The sensitivity of this method was sufficient to detect 5 ng of purified mackerel DNA mixed with 50 ng of surimi DNA. This rapid and specific method for the detection of allergenic mackerel would be beneficial in reducing food allergy caused by the ingestion of hidden allergen in processed food.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2012.05a
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• pp.913-915
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• 2012

RNA polymerase beta subunit gene (rpoB) mutation of rifampin-resistant Mycobacteria was analyzed using nucleotide sequence of rpoB DNA (351 bp) containing rifampin resistant region, $rif^r$. For this purpose, we collected rifampin-resistant Mycobacteria that were identified by conventional culture method from Masan National Hospital and The Korean Institute of Tuberculosis and performed analysis of nucleotide sequence of rpoB of them. We found various mutations of rpoB linked rifampin resistant gene from rifampin-resistant Mycobacteria. From this study, we identified mutations of different codons from codons that have been reported recently.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.24
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• pp.10585-10589
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• 2015

Breast cancer susceptibility gene 1 (BRCA1), mapped on chromosome 17q21, is implicated in the mechanisms of cellular DNA repair. Inactivation of this gene is involved in the development of many human cancers, including breast cancer. This study aimed to investigate the prognostic value of BRCA1 promoter hypermethylation and expression in breast cancer cases. Sixty-one breast cancers were examined for BRCA1 hypermethylation by methylation-specific polymerase chain reaction (PCR), and 45 paired normal breast tissues were analyzed for altered BRCA1 mRNA levels by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Aberrant methylation status in BRCA1 was detected in 15 of 61 cases (24.6%), while reduced expression was found in 7 of 45 (15.6%). BRCA1 hypermethylation was statistically associated with tumor grade III (p=0.04), a high frequency of stage IIB (p=0.02), and triple-negative phenotype (OR= 3.64, 95%CI =1.1-12.3, p=0.03). Our findings indicated that BRCA1 promoter hypermethylation is a useful prognostic marker for breast cancer.

• Tuberculosis and Respiratory Diseases
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• v.43 no.1
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• pp.30-37
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• 1996

Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.

• The Plant Pathology Journal
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• v.17 no.6
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• pp.329-335
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• 2001

Yellowing disease of palms caused by phytoplasma is spreading in the Arabian Gulf region. Surveys were conducted to determine the occurrence of the disease. Electron and fluorescence microscopy, and polymerase chain reaction (PCR) techniques were used to detect the phytoplasma associated with the yellowing disease of ornamental palm Washingtonia sp. grown in Kuwait. An accumulation of phytoplasmal DNA was observed by fluorescence microscopy in phloem tissues of diseased palms. Electron microscopy showed that phytoplasma cells were primarily confined to the phloemsieve elements of tissue samples collected from infected mature palms in the field. The pathogen was identified on the basis of molecular analysis using universal and specific nested primers in PCR amplifications. Prokaryotic 16S rDNA gene was detected in amplified PCR products. Nested PCR resulted in DNA amplification of 1.2 kbp fragment. This is the first report of a phytoplasmal rDNA gene identified from the putative causal pathogen of yellows in ornamental palms in the Arabian Gulf region.