• The Plant Pathology Journal
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• 제36권5호
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• pp.497-502
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• 2020

Barley yellow dwarf virus (BYDV) is an economically important plant pathogen that causes stunted growth, delayed heading, leaf yellowing, and purple leaf tip, thereby reducing the yields of cereal crops worldwide. In the present study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed for the detection of BYDV in oat leaf samples. The RT-RPA assay involved incubation at an isothermal temperature (42℃) and could be performed rapidly in 5 min. In addition, no cross-reactivity was observed to occur with other cereal-infecting viruses, and the method was 100 times more sensitive than conventional reverse transcription polymerase chain reaction. Furthermore, the assay was validated for the detection of BYDV in both field-collected oat leaves and viruliferous aphids. Thus, the RT-RPA assay developed in the present study represents a simple, rapid, sensitive, and reliable method for detecting BYDV in oats.

• The Plant Pathology Journal
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• 제38권4호
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• pp.417-422
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• 2022

Apple stem grooving virus (ASGV) is a destructive viral pathogen of pome fruit trees that causes significant losses to fruit production worldwide. Obtaining ASGV-free propagation materials is essential to reduce economic losses, and accurate and sensitive detection methods to screen ASGV-free plantlets during in vitro propagation are urgently necessary. In this study, ASGV was sensitively and accurately quantified from in vitro propagated apple plantlets using a reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assay. The optimized RT-ddPCR assay was specific to other apple viruses, and was at least 10-times more sensitive than RT-real-time quantitative PCR assay. Furthermore, the optimized RT-ddPCR assay was validated for the detection and quantification of ASGV using micropropagated apple plantlet samples. This RT-ddPCR assay can be utilized for the accurate quantitative detection of ASGV infection in ASGV-free certification programs, and can thus contribute to the production of ASGV-free apple trees.

• 생명과학회지
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• 제13권2호
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• pp.197-205
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• 2003

미국 EPA (Environment Protection Agency)와 환경부에서 규정하고 있는 표준검출법인 총세포배양법 (Total Culturable Virus Assay : TCVA)을 이용하여 부산시 상수원수 및 정수장을 대상으로 2001년 7월에서 2002년 11월까지 바이러스 검사를 실시하였다. 검사결과 21개의 상수원수 시료 중 13개 시료에서 CPE (cytopathic effect)가 확인되어 61.9%의 양성률을 보였으며, 정수 및 수도꼭지수에서는 모두 검출되지 않았다. 또한, 검출된 바이러스를 계절별로 보면 주로 여름철과 초겨울에 분포하는 특성을 보였다. 검출된 바이러스는 TCVA-MPN 방법에 의해 1.92-9.70 MPN/100 L의 범위로 전량 되었으며, 면역형광법 (Immunofluorescent assay)에 의해 human poliovirus type 1과 enterovirus종으로 동정되었다.

• 약학회지
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• 제43권4호
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• pp.469-473
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• 1999

EA, the water soluble substance, was prepared from the carpophores of Elfvingia applanata (Pers). Karst. Anti-influenza A virus (anti-Flu A) activity of EA was examined of Vero cells by plaque reduction assay in vitro. And the combined antiviral effects fo EA with interferon (IFN) alpha and gamma were examined on the multiplication of Flu A with 50% effective concentration ($EC_50$) of 1.50 mg/ml. The results of combination assay were evaluated by the combination index (CI) that was analysed by the multiple drug effect analysis. The combination of EA with IFN alpha on Flu A showed more potent synergism with CI values of 0.50~0.52 of 50%, 70%, 90% effective levels than that with IFN gamma with CI values of 0.82~0.99.

• 생약학회지
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• 제31권1호
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• pp.82-86
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• 2000

To evaluate anti-influenza virus activity of 113 specimens of Korean traditional medicine both water and methanol extracts were examined using haemagglutination inhibition test. The water extract from Citrus junos was found to inhibit influenza virus A/Taiwan/l/86(H1N1). The survival rates of virus were determined by in situ cellular enzyme-linked immunosorbent assay. The water extract of Citrus junos was fractionated by chromatographic separating using Amberlite XAD-4, 40% MeOH and 60% MeOH layer had antiviral activity. The half inhibition concentration $(IC_{50})$ of 40% MeOH layer on survival of influenza virus was $MIC>361.5{\mu}g/ml$ and $IC_{50}$ value of fr. 40-4 fractionated from 40% MeOH layer was $677.19{\mu}g/ml$. These results suggested that the fractions of Citus junos have potent anti-influenza A virus activity.

• Journal of Microbiology and Biotechnology
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• 제22권8호
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• pp.1165-1169
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• 2012

In April 2009, the H1N1 pandemic influenza virus emerged as a novel influenza virus. The aim of this study was to compare the performances of several molecular assays, including conventional reverse transcription polymerase chain reaction (RT-PCR), two real-time reverse transcription (rRT)-PCRs, and two multiplex RTPCRs. A total of 381 clinical specimens were collected from patients (223 men and 158 women), and both the Seeplex RV7 assay and rRT-PCR were ordered on different specimens within one week after collection. The concordance rate for the two methods was 87% (332/381), and the discrepancy rate was 13% (49/381). The positive rates for the molecular assays studied included 93.1% for the multiplex Seeplex RV7 assay, 93.1% for conventional reverse transcription (cRT)-PCR, 89.7% for the multiplex Seeplex Flu ACE Subtyping assay, 82.8% for protocol B rRT-PCR, and 58.6% for protocol A rRT-PCR. Our results showed that the multiplex Seeplex assays and the cRT-PCR yielded higher detection rates than rRT-PCRs for detecting the influenza A (H1N1) virus. Although the multiplex Seeplex assays had the advantage of simultaneous detection of several viruses, they were time-consuming and troublesome. Our results show that, although rRT-PCR had the advantage, the detection rates of the molecular assays varied depending upon the source of the influenza A (H1N1)v virus. Our findings also suggest that rRT-PCR sometimes detected virus in extremely low abundance and thus required validation of analytical performance and clinical correlation.

• 한국균학회지
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• 제27권2호통권89호
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• pp.175-179
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• 1999

잔나비걸상 Elfvingia applananta 자실체의 수용성물질 EA의 vesicular stomatitis virus[New Jersey serotype, VSV(NJ)]에 대한 항바이러스효과를 plaque reduction assay에 따라 실험한 결과 EA는 용량의존적으로 plaque 형성을 억제하였으며 $EC_{50}$는 2.10 mg/ml이었다. 단백질성 항바이러스제인 interferon(IFN)과 EA와의 병용시험 결과 f(a)가 0.50에서 0.90인 유효농도범위에서 IFN alpha와 병용시 f(a)의 값이 커짐에 따라 상가작용 내지는 길항작용을 나타내었으며, IFN gamma와의 병용시에는 길항작용을 나타내었다. 따라서 IFN alpha와의 병용시 f(a)가 0.50내지 0.70의 유효농도 범위에서 상가효과가 있다.

• 미생물학회지
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• 제38권4호
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• pp.255-255
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• 2002

For rapid and sensitive measurement of antiviral activities, application of a recombinant virus containing firefly luciferase gene was attempted. Recombinant human cytomegalovirus (HCMV) containing luciferase gene driven by HCMV late gene pp28 promoter (HCMV/pp28-luc) was used to test the antiviral activities of three known compounds and the result was compared with results from the conventional plaque assay for measuring the production of infectious viruses. When human fibroblast cells were infected with HCMV/pp28-luc, luciferase activity was observed at 2 days after infection and reached maximum at 6 days after infection, whereas the production of infectious virus was maximal at 4 days after infection. The antiviral activities of ganciclovir, acyclovir, and papaverine were measured in HFF cells infected with HCMV/PP28-luc and the luciferase activity was compared with the infectious virus titers. Luciferase activity decreased as the concentration of ganciclovir or papaverine increased, while there was a slight decrease in luciferase activity with acyclovir. The level of the decrease in Luciferase activity was comparable to the level of decrease in the production of infectious virus. Therefore, the antiviral assay using recombinant virus HCMV/pp28-luc resulted in sensitivity similar to the conventional plaque assay with a significant reduction in assay time.

• IMMUNE NETWORK
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• 제21권6호
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• pp.39.1-39.18
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• 2021

The high virulent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that emerged in China at the end of 2019 has generated novel coronavirus disease, coronavirus disease 2019 (COVID-19), causing a pandemic worldwide. Every country has made great efforts to struggle against SARS-CoV-2 infection, including massive vaccination, immunological patients' surveillance, and the utilization of convalescence plasma for COVID-19 therapy. These efforts are associated with the attempts to increase the titers of SARS-CoV-2 neutralizing Abs (nAbs) generated either after infection or vaccination that represent the body's immune status. As there is no standard therapy for COVID-19 yet, virus eradication will mainly depend on these nAbs contents in the body. Therefore, serological nAbs neutralization assays become a requirement for researchers and clinicians to measure nAbs titers. Different platforms have been developed to evaluate nAbs titers utilizing various epitopes sources, including neutralization assays based on the live virus, pseudovirus, and neutralization assays utilizing recombinant SARS-CoV-2 S glycoprotein receptor binding site, receptor-binding domain. As a standard neutralization assay, the plaque reduction neutralization test (PRNT) requires isolation and propagation of live pathogenic SARS-CoV-2 virus conducted in a BSL-3 containment. Hence, other surrogate neutralization assays relevant to the PRNT play important alternatives that offer better safety besides facilitating high throughput analyses. This review discusses the current neutralization assay platforms used to evaluate nAbs, their techniques, advantages, and limitations.

• 생약학회지
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• 제30권3호
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• pp.244-249
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• 1999

In order to find less toxic antiviral agents from Basidiomycetes, EA, the water soluble substance, was prepared from the carpophores of Elfvingia applanata(Pers.) Karst. Antiviral activity of EA against vesicular stomatitis virus [Indiana serotype, VSV(IND)] was examined in Vero cells using plaque reduction assay in vitro. And the combined antiviral effects of EA with interferon (IFN) alpha or gamma were examined on the multiplication of VSV(IND). EA caused a concentration-dependent reduction in the plaque formation of VSV(IND) with 50% effective concentration $(EC_{50})$ of $104.02\;{\mu}g/ml$. The results of combination assay were evaluated by the combination index (CI) that was analysed by the multiple drug effect analysis. All cases of the combination of EA with IFN alpha or IFN gamma showed potent synergism with CI values of $0.38{\sim}0.52$ for $50{\sim}90%$ effective levels.