• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2002.11a
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• pp.143-143
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• 2002

돼지 장염 바이러스를 동시에 감별 진단할 수 있는 새로운 진단법인 oligonicleotide microarray 기법을 돼지 설사 분변을 대상으로 바이러스 감염을 진단하고 기존의 진단법으로 널리 사용되고 있는 RT-PCR과 진단기법의 민감도와 특이도를 비교하고자 하였다. 32개 양돈장, 102 예의 포유 및 이유자돈의 설사분변에서 microarray를 이용한 검사결과 Transmissible gastroenteritis virus (TGEV) 9.8％, porcine epidemic diarrhea virus (PEDV) 28％, (porcine enteric calicivirus (PECV) 18.6％, porcine rotavirus (PRV) group A 6.9％, PRY group C 1％ 의 검출률을 확인하였다. 또한 RT-PCR 기법과의 비교에서도 100％의 민감도와 72.2％의 특이도를 보였으며 agreement는 85.3％, kappa value 0.71로 우수한 진단기법임을 확인하였다. 이를 통해 microarray 진단법은 RT-PCR 후의 전기영동 과정과 민감도를 높이기 위해 수행되는 nested PCR 수행의 번거러움을 없애면서 정확한 감별진단을 수행할 수 있는 진단 기법임을 확인하였다.

• Journal of fish pathology
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• v.23 no.1
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• pp.17-25
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• 2010

We have identified an iridovirus CH-1 from sea perch Lateolabrax sp. healthy externally and imported from China to Korea. In a comparison of the nucleotide sequences of the five different genomic regions, the CH-1 appears to be closely related to the ISKNV, IVS-1 and Ehime-1 strains detected in China, Korea and Japan respectively. In quantitative comparison of the viral DNA, level of CH-1 in tissue of imported fish was 10,000 times lower than that of IVS-1 strain presented in the infected rock bream Oplegnathus fasciatus of moribund stage. It allowed us to speculate the possibility of the asymtomatic iridovirus infection in the culturing sea perch. Such possibility of asymptomatic infection was supported by result of no appearance of dead fish with typical symptoms of iridoviral disease in keeping experiment of the imported sea pearch in laboratory for more than three weeks. Such asymptomatic infections with iridovirus were also found in spleen of the culturing and externally healthy sea perch of Korea by the presence of the iridoviral DNA in nested PCR.

• Korean Journal of Veterinary Service
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• v.31 no.3
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• pp.273-281
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• 2008

This survey was conducted to investigate the infection of caprine arthritis encephalitis virus (CAEV). From April to October 2007, we examined a total of 173 goats in 7 dairy goat-breeding farms located in Na-ju, Hae-nam, Young-am, and Young-gwang area of Jeonnam province. The results show that 41 (23.7%) goats are positive for CAEV, confirmed by nested PCR of 173 blood samples. Regional analysis revealed that a positive proportion for CAEV was 50.0% in Young-gwang, 25.6% in Young-am, 25.0% in Hae-nam, and 10.9% in Na-ju. Clinical signs were observed in 17 (9.8%) goats, of which 6 suffered from arthritis, 7 from mastitis, and 4 from pneumonia. Among the examined 173 goats, 8(4.6%) goats are positive for CAEV accompanied with CAE symptoms. There was a tendency to be much higher(p<0.05) levels of natrium(Na), chlorine(Cl) in CAEV-positive than in CAEV-negative goats. However, other serum biochemical values were no statistically significant effect.

• 대한생식의학회:학술대회논문집
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• 2007.11a
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• pp.136-136
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• 2007

• Journal of the Korean Society of Tobacco Science
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• v.37 no.1
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• pp.25-33
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• 2015

Genome walking is a par ticular application for identifying sequences of unknown genomic regions adjacent to a known region. Many genome walking methods based on polymerase chain reaction (PCR) are available. Even if earlier techniques suffer from low reproducibility, inefficiency, and non-specificity, improved strategies have been developed. In this study, we present an alternative strategy: the genomic DNA is digested with restriction enzymes. After cytosine overhangs at 5' ends, the fragments are ligated to linker adaptor s had guanine overhang at 3' ends. Then nested PCR is performed. The improvements in this strategy focus on two points. The first is the C tailing method using Pfu polymerase instead of the A tailing method based on nontemplate-dependent terminal transferase activity of Taq polymerase. Therefore unintended modification of target DNA can be prevented without A tailing error. The second point is the use of C/G-specific ligation had advantage in the ligation efficiency compared with A/T-specific ligation. Therefore, the C-G linker PCR method increases ligation efficiency between digested genomic DNA and adaptor DNA. As a result, the quantity of target DNA to amplify by PCR is enriched. We successfully used G-C linker PCR to retrieve flanking regions bordering the phophinothricin resistance gene in genetically modified tobacco (GMO).

• Journal of fish pathology
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• v.34 no.2
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• pp.141-147
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• 2021

In this study, of the imported shrimps between 2017 and 2020, we investigated white spot syndrome virus (WSSV), covert mortality nodavirus (CMNV) and decapod iridescent virus 1 (DIV-1). Of the imported shrimps (a total of 29 groups), WSSV was detected as 31% (9/29) by nested PCR assay. And CMNV and DIV-1 were not identified in this study. To investigate the genetic relatedness of WSSV identified from imported shrimp, VR 14/15 region showed WSSV genomic variable loci was compared with reference isolates. Among the nine WSSV-positive samples, VR 14/15 region was amplified in only a sample (20-CH-1 isolate, imported from China in 2020). And the 20-CH-1 isolate showed 99.8% identity with WSSV-IN-05-01 which was reported in India in 2005, suggesting that those of WSSV have been spread from India to China. Furthermore, although the pathogenicity of WSSV identified from frozen shrimp was not evaluated, the international trade of diseased frozen shrimps could be led to the potential risk of virus transmission.

• BMB Reports
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• v.40 no.2
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• pp.172-179
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• 2007

PCR-mediated recombination describes the process of in vitro chimera formation from related template sequences present in a single PCR amplification. The high levels of genetic redundancy in eukaryotic genomes should make recombination artifacts occur readily. However, few evolutionary biologists adequately consider this phenomenon when studying gene lineages. The cytosolic glyceraldehyde-3-phosphate dehydrogenase gene (GapC), which encodes a NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the cytosol, is a classical lowcopy nuclear gene marker and is commonly used in molecular evolutionary studies. Here, we report on the occurrence of PCR-mediated recombination in the GapC gene family of Hibiscus tiliaceus. The study suggests that recombinant areas appear to be correlated with DNA template secondary structures. Our observations highlight that recombination artifacts should be considered when studying specific and allelic phylogenies. The authors suggest that nested PCR be used to suppress PCRmediated recombination.

• Fisheries and Aquatic Sciences
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• v.7 no.3
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• pp.122-129
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• 2004

Harmful Cochlodinium polykrikoides is a notorious harmful algal bloom (HAB) species that is causing mass mortality of farmed fish along the Korean coast with increasing frequency. We analyzed the sequence of the large subunit (LSD) rDNA D1-D3 region of C. polykrikoides and conducted phylogenetic analyses using Bayesian inference of phylogeny and the maximum likelihood method. The molecular phylogeny showed that C. polykrikoides had the genetic relationship to Amphidinium and Gymnodinium species supported only by the relatively high posterior probabilities of Bayesian inference. Based on the LSU rDNA sequence data of diverse dinoflagellate taxa, we designed the C. polykrikoides-specific PCR primer set, CPOLY01 and CPOLY02 and developed PCR detection assays for its sensitive, accurate HAB monitoring. CPOLY01 and CPOLY02 specifically amplified C. polykrikoides and did not cross-react with any dinoflagellates tested in this study or environmental water samples. The effective annealing temperature $(T_{p})$ of CPOLY01 and CPOLY02 was $67^{\circ}C$. At this temperature, the conventional and nested PCR assays were sensitive over a wide range of C. polykrikoides cell numbers with detection limits of 0.05 and 0.0001 cells/reaction, respectively.

• Biomedical Science Letters
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• v.16 no.1
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• pp.10-18
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• 2010

The present study was set to develop comprehensive system for assessing the safety of drinking water using PCR-reverse blot hybridization assay (REBA). The REBA developed in this study can detect waterborne pathogens such as Shigella spp., Salmonella spp., Citrobacter spp., Enterobacter spp., Klebsiella spp., Yersinia spp., Mycobacterium spp., Listeria spp. at the genus level, and Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Pseudomonas aeruginosa, Yersinia enterocolitica, Y. pseudotuberculosis, Mycobacterium avium complex, M. marinum, Enterococcus faecalis, and Staphylococcus aureus at the species level, and E. coli O157:H7 at the strain level.

• Korean Journal of Veterinary Service
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• v.46 no.3
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• pp.235-241
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• 2023

Noroviruses are a major cause of gastroenteritis in humans and animals worldwide. In 2021, canine norovirus (CNoV) infection was detected at an animal clinic in Gwangju area, South Korea. A semi-nested polymerase chain reaction was developed to amplify a 478 bp fragment of the RdRp gene of CNoV. The phylogenetic analysis of this fragment confirmed the strain to be genogroup IV.2 (Dog/GIV.2/gw/s377/2021/KOR), which exhibited the highest similarity to the feline NoV strain GIV.2/CU081210E/USA/2010 (accession no. NC_045762) with 95.1% nucleotide (nt) identity and 98.7% amino acid (aa) identity. These research findings indicate that the detected norovirus in dogs is genetically similar to a feline-origin norovirus, suggesting easy cross-species transmission among animals.