• Journal of Apiculture
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• v.32 no.3
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• pp.171-180
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• 2017

Slow Bee Paralysis Virus (SBPV) is a pathogenic virus against honeybee and bumblebee, causes the death of adult bee by paralyzing the fore-leg of bee. In this study, for rapid detection of SBPV from bumblebee, SBPV-specific Ultra-rapid Reverse transcription PCR was developed. After optimizing of SBPV-specific Ultra-rapid PCR, the existence of $1.0{\times}10^8$ SBPV-specific DNA molecules could be recognized in 3 minute and 35 seconds. Even $1.0{\times}10^1$ molecules of SBPV-specific DNA could be measured with quantitative manner. Meanwhile, from both imported bumblebee and bumblebee produced in Korea, SBPV were detected using proposed method. In the laboratory as well as in the field, SBPV-specific Ultra-rapid Reverse transcription PCR would be applied and might be expected as useful tools at production of bumblebee or inspection for the import and export system of bumblebee.

• Parasites, Hosts and Diseases
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• v.54 no.2
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• pp.239-241
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• 2016

Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

• Journal of the Korea Institute of Military Science and Technology
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• v.20 no.5
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• pp.726-732
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• 2017

Bacillus anthracis, Vibrio cholerae, Variola virus and Shigella dysenteriae are classified as category A and B biological weapons. In this study suggest that 4 genes of Bacillus anthracis, 2 genes of Vibrio cholerae, 1 gene of Variola virus and 1 gene of Shigella dysenteriae were detective 50~500 fg of target DNA per reaction using real-time PCR based assay. Also analytical specificity did not show any cross-reactivity with other related bacteria. Reliable and one reaction could be effective early diagnostic and treatment for detection of unknown samples.

• The Plant Pathology Journal
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• v.33 no.1
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• pp.34-42
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• 2017

Grapevine rupestris stem pitting-associated virus (GRSPaV) is a worldwide-distributed pathogen in grapevines with a high genetic variability. Our study revealed differences in the complexity of GRSPaV population in a single host. A single-variant GRSPaV infection was detected from the SK30 grapevine plant. On the contrary, SK704 grapevine was infected by three different GRSPaV variants. Variant-specific RT-PCR detection protocols have been developed in this work to study distribution of the three different variants in the same plant during the season. This study showed their randomized distribution in the infected SK704 grapevine plant. Comparative analysis of full-length genome sequences of four Slovak GRSPaV isolates determined in this work and 14 database sequences showed that population of the virus cluster into four major phylogenetic lineages. Moreover, our analyses suggest that genetic recombination along with point mutations could play a significant role in shaping evolutionary history of GRSPaV and contributed to its extant genetic diversification.

• The Plant Pathology Journal
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• v.17 no.6
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• pp.325-328
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• 2001

Gelatin particle agglutination test (GPAT) was used to detect Cymbidum mosaic virus (CymMV) in Cattleya plants. Gelatin particles were coated with purified anti-CymMV immunoglobulin of 25-100 $\mu\textrm{g}$/ml and were subjected to several different concentrations of purified CyMfV as well as varying dilutions of orchid leaf extracts. The GPAT detected purified CymMV up to a minimum concentration of 10 $\mu\textrm{g}$/ml. CymMV was detected from crude sap extract of infected Cattleya leaves and roots up to 1:51,200 and 1:25,600 dilutions, respectively. However, the optimum range of leaf and root sap dilutions was between 50-100. Non-specific reactions were not encountered from any of the healthy orchid plants tested. The entire GPAT process was completed within 2-3 hours. This test was found to be very useful for the detection of CymMV in orchids because it is sensitive, economical, and easy to perform.

• Microbiology and Biotechnology Letters
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• v.46 no.3
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• pp.269-276
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• 2018

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.541-548
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• 1989

The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

• Korean Journal of Veterinary Research
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• v.28 no.2
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• pp.365-369
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• 1988

The present study was done to demonstrate ADV antigens in frozen and paraffin sections from ADV-infected pigs and cell cultures by using of the IGS method. Tissue specimens from 3 young pigs infected with ADV-phylaxia strain and of 2 healthy pigs were used. Fibroblastic cells originated from pig brain and BHK cells were grown and confluent monolayers were infected with the virus. Two monoclonal antibodies and a specific hyperimmune serum to ADV were used as the source of primary antibodies for both the IGS and immunoperoxidase methods. Application of the IGS method yielded a black fine granular reaction in positive areas, and the results were superior to those obtained using the immunoperoxidase technique for all cases tested. The IGS method might be useful in the detection of various viral antigens in tissue sections.

• Korean Journal Plant Pathology
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• v.13 no.4
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• pp.219-224
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• 1997

A PT-PCR (reverse transcription-polymerase chain reaction) diagnostic method for potato virus Y (PVY) was developed using primer pair derived from conserved region of coat protein genes of several PVY strains, A 764 bp PCR product was detected from several lines of potato cv. Atlantic. We could prove that the 764 bp DNA fragment was indeed the PVY gene by sequencing analysis. PVY detection method using RT-PCR technique was about tuber tissue.

• Research in Plant Disease
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• v.16 no.2
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• pp.121-124
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• 2010

Immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) was applied to the detection of Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) on Cucurbitaceae crops. These seed-borne tobamoviruses were accurately detected from the infected leaves and seeds by IC-RT-PCR. This method was estimated to be about 100 times more sensitive than ELISA, and also it allowed the direct confirmation of ELISA results by using the captured antigens from a completed ELISA microwell. This convenient and reliable method could be used routinely for large-scale field surveys or seed tests of Cucurbitaceae crops.