• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.86-93
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• 2000

Paratuberculosis (Johne's disease), a chronic enteritis produced by Mycobacterium paratuberculosis, affects a large proportion of ruminants in all continents and causes important economic losses. The identification of well-characterized and species-specific components of M paratuberculosis would provide the means to improve the specificity and sensitivity of immunodiagnostic assays for Johne's disease. The aims of this study were to express the recombinant C-terminal of 34kDa protein (rC34P) of M paratuberculosis in E coli and to investigate the effectiveness of this protein in detecting antibodies to the native protein in sera from paratuberculosis infected cattle. The C-terminal of the gene encoding the 34kDa protein was amplified by polymerase chain reaction from the chromosomal DNA of M paratuberculosis (ATCC 19698) and cloned into vector pGEX-4T-2. Then, cloned plasmid was transformed into E coli DH5${\alpha}$ and the rC34P was overexpressed. The rC34P was purified by affinity chromatography and gel filtration. The rC34P was examined antigenicity by Western blot. The rC34P was reactive with culture positive bovine serum and hyperimmune rabbit anti-M paratuberculosis serum but was not reactive with culture negative bovine serum and tuberculin positive bovine serum in Western blot. In conclusion, the rC34P produced in this study is expected as a useful candidate for antigen in serological diagnosis of Johne's disease.

• The Plant Pathology Journal
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• v.18 no.6
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• pp.323-327
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• 2002

A rod-shaped virus was isolated from pepper showing mild mosic during the winter growing seasons of 2001 and 2002 in Korea. Based on its biological reactions, serological relationships, reverse transcription-poly-merase chain reaction (RT-PCR) using specific primers, and nucleotide sequence analysis of coat protein (CP) gene, the isolated virus was identified as Tobacco mild green mosaic virus (TMGMV) and designated as Korean pepper isolate (TMGMV-KP). Crude sap from infected tissue was mechanically transmitted to various indicator plants, which produced characteristic symptoms of tobamovirus infection. However, no symptom was observed in Gomphorena globosa. In RT-PCR assays with specific primers toy respective detection of TMGMV, Tobacco mosaic virus (TMV), Pepper mild mottle virue (PMMoV), and Tomato mosaic virus (ToMV), a single strong band of about 500 bp in length was produced from the sample used only with TMGMV primers. The amplified DNA was cloned and the nucleotide sequence was determined. Sequence comparisons with the CP gene of other tobamoviruses indicated that TMGMV-KP shared 99.3％ identity with TMGMV Japanese isolate and only 59.1, 58.6, and 58.1％ identity with TMV, PMMoV and ToMV, respectively. This is the first report of TMGMV in Korea.

• The Plant Pathology Journal
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• v.33 no.4
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• pp.382-392
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• 2017

This study reports the findings of a distinct Potato virus Y (PVY) isolate found in Northeast China. One hundred and ten samples (leaves and tubers) were collected from potato plants showing mosaic symptoms around the city of Harbin in Heilongjiang province of China. The collected tubers were planted and let to grow in a greenhouse. New potato plants generated from these tubers showed similar symptoms, except for one plant. Subsequent serological analyses revealed PVY as the causing agent of the disease. A novel PVY isolate (referred to as HLJ-C-44 in this study) was isolated from this sample showing unique mild mosaic and crisped leaf margin symptoms. The complete genome of this isolate was analyzed and determined. The results showed that HLJ-C-44 is a typical PVY isolate. Phylogenetic analysis indicated that this isolate belongs to the N-Wi strain group of PVY recombinants ($PVY^{N-Wi}$) and also shared the highest overall sequence identity (nucleotide and amino acid) with other members of this strain group. However, recombination analysis of isolate HLJ-C-44 revealed a recombination pattern that differed from that of other $PVY^{N-Wi}$ isolates. Moreover, biological assays in four different potato cultivars and in Nicotiana tabacum also revealed a different phenotypic response than that of a typical $PVY^{N-Wi}$ isolate. This data, combined, suggest that HLJ-C-44 is a novel PVY recombinant with distinct biological properties.

• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.418-427
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• 2023

Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

• Journal of Life Science
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• v.30 no.8
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• pp.731-741
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• 2020

Coronavirus disease 19 (COVID-19) is caused by SARS-CoV-2 (Severe Acute Respiratory SyndromeCoronavirus 2). To date, seven coronaviruses that can infect humans were reported. Among them, infections with four coronavirus strains (HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1) resulted in mild symptoms such as common cold, whereas SARS-CoV and MERS-CoV caused severe symptoms and epidemics in 2002 and 2012, respectively. In the most recent, SARS-CoV-2 was first reported in Wuhan, China in December 2019 and became a notorious cause of the ongoing global pandemics. To diagnose, treat, and prevent COVID-19, the development of rapid and accurate diagnostic tools, specific therapeutic drugs, and safe vaccines essentially are required. In order to develop these powerful tools, it is prerequisite to understand a phenotype, a genotype, and life cycle of SARS-CoV-2. Diagnostic techniques have been developing rapidly around world and many countries take the fast track system to accelerate approval. Approved diagnostic devices are rapidly growing facing to urgent demand to identify carriers. Currently developed commercial diagnostic devices are divided into mainly two categories: molecular assay and serological & immunological assay. Molecular assays begins the reverse transcription step following polymerase chain reaction or isothermal amplification. Immunological assay targets SARS-CoV-2 antigen or anti-SARS-CoV-2 antibody of samples. In this review, we summarize the phenotype, genome structure and gene expression of SARS-CoV-2 and provide the knowledge on various diagnostic techniques for SARS-CoV-2.

• Tuberculosis and Respiratory Diseases
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• v.47 no.3
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• pp.311-320
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• 1999

Background: In recent years, tuberculosis has re-emerged as a major health problem in both industrialized & developing countries. Recent advances in identifying & purifying antigens secreted in active tuberculosis infection have lead to the development of serological assays based on a number of immunodominant antigens. To date, the most sensitive and specific of these antigens has been the 38-kDa antigen. Method: Two rapid membrane-based serologic assays using antigen(38-kDa) from mycobacterium tuberculosis for the diagnosis of tuberculosis were evaluated in 22 patients with smear-positive pulmonary tuberculosis, 14 patients with inactive pulmonary tuberculosis, and 9 patients with non-tuberculous lung disease. Result: The evaluation of validity(sensitivity, specificity, positive predictive value, negative predictive value, false positivity and false negativity) of STAT-PAK ULTRA FAST$^{(R)}$ were 77.3%, 28.6%, 63.0%, 44.4%, 71.4 %, and 22.7% for differential diagnosis of active pulmonary tuberculosis and inactive pulmonary tuberculosis, respectively. The evaluation of validity of STAT-PAK ULTRA FAST$^{(R)}$ were 77.3%, 33.3%, 73.9%, 37.5%, 66.7%, and 22.7% for differential diagnosis of active pulmonary tuberculosis and non-tuberculosis. The evaluation of validity of ICT Tuberculosis$^{(R)}$ were 54.5%, 57%, 66.7%, 44.4%, 42.9%, and 45.5% for differential diagnosis of active pulmonary tuberculosis and inactive pulmonary tuberculosis. The evaluation of validity of ICT Tuberculosis$^{(R)}$ were 54.5%, 100%, 100%, 47.4%, 0%, and 45.4% for differential diagnosis of active pulmonary tuberculosis and non-tuberculosis. Conclusion: We concluded no effectiveness of STAT-PAK ULTRA FAST$^{(R)}$ & ICT tuberculosis$^{(R)}$on serologic diagnosis of pulmonary tuberculosis. In the future, further large-scale study should be needed for serologic diagnosis of pulmonary tuberculosis.

• Journal of Life Science
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• v.23 no.10
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• pp.1267-1272
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• 2013

The aim of this study was to determine the prevalence and infection status of Salmonella species (spp.) in 25 conventional pig farms by traditional fecal culture and serological methods to develop a Salmonella control program for Korean pig farms. The individual seroprevalence of Salmonella spp. in pigs reared in the 25 pig farms was 83.1% in sows and 6.4-32% in different aged pig groups, with the total seroprevalence 28.4% (141/848). The seroprevalence of the tested pigs increased in accordance with the decrease in maternal antibody and the rearing period on these farms. Of note, all the 25 pig farms contained at least two or more anti-Salmonella antibody-positive sows. In the fecal cultures Salmonella spp. were isolated only in three (12.0%, 3/25) of 16 serologically Salmonella-suspected farms (64.0%, 16/25), showing the limitation of the fecal culture method and the need for serum assays to understand the exact status of Salmonella infection in swine herds, which likely contain subclinically infected pigs or carriers. The results highlight the need to establish a supply system of Salmonella-free gilts for the promotion of a national Salmonella control program on swine farms in Korea. Further studies will be needed to develop an effective monitoring system for the implementation of a national Salmonella control program.