• 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.

• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.210-217
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• 2018

A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of the developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Moreover, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with the four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.

• BMB Reports
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• 제37권5호
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• pp.546-551
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• 2004

The increasing pace of development in molecular biology during the last decade has had a direct effect on mass testing and diagnostic applications, including blood screening. We report the model Microarray that has been developed for Hepatitis B virus (HBV) and Hepatitis D virus (HDV) detection. The specific primer pairs of PCR were designed using the Primer Premier 5.00 program according to the conserved regions of HBV and HDV. PCR fragments were purified and cloned into pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was prepared by robotically spotting PCR products onto the surface of glass slides. Sequences were aligned, and the results obtained showed that the products of PCR amplification were the required specific gene fragments of HBV, and HDV. Samples were labeled by Restriction Display PCR (RD-PCR). Gene chip hybridizing signals showed that the specificity and sensitivity required for HBV and HDV detection were satisfied. Using PCR amplified products to construct gene chips for the simultaneous clinical diagnosis of HBV and HDV resulted in a quick, simple, and effective method. We conclude that the DNA microarray assay system might be useful as a diagnostic technique in the clinical laboratory. Further applications of RD-PCR for the sample labeling could speed up microarray multi-virus detection.

• 식물병연구
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• 제7권3호
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• pp.170-174
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• 2001

시판고추 종자로부터 Tobamovirus를 검출하고 Tobamovirus 오염률을 조사하고자 gelatin particle agglutination test (GPAT)의 최적조건을 조사하였다. Gelatin 입자에 감작할 담배 모자이크 바이러스 고추 계통에 대한 ${\gamma}$-globulin G 의 최적농도는 100$\mu\textrm{g}$/$m\ell$이었고, GPAT의 바이러스 검출 감도는 효소면역항체법 및 dot immuno-binding assay와 유사하였으며, Tobamovirus 검출을 위한 종자시료의 최적 희석배수는 5배에서 25배였다. 그리고 최적화된 GPAT에 의한 국내시판 고추종자의 Tobamovirus 오염률은 평균 79.1%인 것으로 판명되었다.

• 한국정보과학회논문지:정보통신
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• 제29권6호
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• pp.663-673
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• 2002

서버 수준의 안티바이러스는 특정 도메인 내에 진입하는 악성코드를 진입점에서 감지하므로 모든 클라이언트를 완벽하게 통제하기 어려운 실제 상황에서 전자우편 서버 등에 유용하게 사용된다. 그러나, 알려지지 않은 악성 코드에 감지에 유용한 행위 감시 기법은 서버에 적용이 어려우므로, 현재의 서버용 안티바이러스들은 이미 알려진 악성 코드에 대한 시그너쳐 기반의 감지, 단순한 필터링 그리고 파일명 변경과 같은 기능만을 수행한다. 본 논문에서는 서버에서의 실행만으로 별도의 안티바이러스가 탑재되지 않은 클라이언트에서도 지속적인 행위 감시가 가능하도록 하는 악성 스크립트 감지 기법을 제안하고 그 구현에 관해 기술한다.

• The Plant Pathology Journal
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• 제21권4호
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• pp.343-348
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• 2005

The partial nucleotide sequences of the genomic dsRNA mycoviruses infecting Pleurotus ostreatus (isolates ASI2596, ASI2597, and Bupyungbokhoe) and Agaricus blazei Murrill were determined and compared with those of the other dsRNA mycoviruses. Partial nucleotide sequences of the purified dsRNA from ASI2596 and ASI2597 revealed RNA-dependent RNA polymerase sequences that are closely related to Oyster mushroom isometric virus 2, while nucleotide sequences and the deduced amino acid sequence from dsRNA mycovirus infecting Agaricus blazei did not show any significant homology to the other dsRNA mycoviruses. Specific primers were designed for RT-PCR detection of these dsRNA viruses and were found to specifically detect each dsRNA virus. Northern blot analysis confirmed the homogeneity of RT-PCR products to each purified dsRNA. Altogether, our results suggest that these virus-specific primer sets can be employed for the specific detection of each dsRNA mycovirus in infected mushrooms.

• Journal of Veterinary Science
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• 제21권4호
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• pp.68.1-68.8
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• 2020

A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.

• Pediatric Infection and Vaccine
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• 제21권3호
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• pp.207-213
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• 2014

목적: 본 연구는 소아에서 호흡기바이러스 감염과 폐렴구균의 상기도 보균율 간의 연관성을 분석하고자 하였다. 방법: 2009년 5월부터 2010년 6월까지 서울대학교 어린이병원에 호흡기 증상을 주소로 내원한 18세 미만 소아로부터 채취한 비인두 흡인물을 대상으로 폐렴구균을 배양하고 RT-PCR을 통해 호흡기 바이러스(influenza virus A와 B, parainfluenza virus 1, 2와 3, respiratory syncytial virus A와 B, adenovirus, rhinovirus A/B, human metapneumovirus, human coronavirus 229E/NL63, OC43/HKU1)를 검출하여 호흡기바이러스 검출과 폐렴구균 보균 사이의 연관성을 분석하였다. 결과: 대상 환자의 중앙 연령은 27개월이었다. 총 1,367건의 비인두 흡인물 중 폐렴구균이 배양된 검체는 228개(16.7%)이었고, 호흡기바이러스가 검출된 검체는 731개(53.5%)이었다. 흔히 분리된 바이러스는, rhinovirus(18.4%), respiratory syncytial virus (RSV) A (10.6%), adenovirus (6.9%), influenza virus A (6.8%) 순으로 나타났다. 폐렴구균 보균율은 호흡기바이러스 양성인 경우가 21.3% (156/731)로 음성인 경우 11.3% (72/636)보다 높았다(P<0.001). 검출된 호흡기바이러스의 종류에 따라서는 influenza virus A [24.7% (23/93) vs 16.1% (205/1274), P=0.001], RSV A [28.3% (41/145) vs 15.3% (187/1222), P=0.001], RSV B [31.3% (10/32) vs 16.3% (218/1335), P=0.042], rhino-virus A/B [22.6% (57/252) vs 15.3% (171/1115), P=0.010]가 양성인 소아는 음성인 소아에 비하여 폐렴구균 보균율이 높게 나타났다. 결론: 본 연구 결과, 호흡기 증상이 있는 소아에서 호흡기바이러스가 검출된 경우 폐렴구균 보균율이 높았다. 향후 호흡기바이러스와 폐렴구균의 보균에 의한 호흡기 감염병의 임상발현 기전을 밝히는 데 도움이 될 것으로 생각된다.

• 대한수의학회지
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• 제47권4호
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• pp.399-407
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• 2007

Cause of high lethality and dissemination to human being, new development of rapid method for the detection of highly pathogenic Avian Influenza Virus (AIV) is still necessary. For the detection of AIV subtype H5N1, typical pathogenic AIV, new method to confirm sub-typing of this virus is also needed. For the purpose of ultra-rapid detection and sub-typing of hemagglutinin and neuraminidase of AIV, this study was planned. As the results we could demonstrate an ultra-rapid multiplex real-time PCR (URMRT PCR) for the detection of AIV In this study, the URMRT PCR were optimized with synthesized AIV H5- and AIV Nl-specific DNA templates and GenSpector TMC, which is a semiconductor process technology based real-time PCR system with high frequencies of temperature monitoring. Under eight minutes, the amplifications of two AIV subtype-specific PCR products were successfully and independently detected by 30 cycled ultra-rapid PCR, including melting point analysis, from $1{\times}10^3$ copies of mixed template DNA. The URMRT PCR for the detection of AIV H5N 1 developed in this study could be expected to apply not only detections of different AIVs, but also various pathogens. It was also discussed that this kind of the fastest PCR based detection method could be improved by advance of related technology in near future.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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• pp.353.2-353.2
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• 2016

We studied electrochemical detection of Botulinum neurotoxin, Vaccinia virus, and Streptococcus Pneumoniae based on nanogap device. Target bio substances were employed as representative targets of protein, virus, and bacteria, respectively. Redox current generated by ferri/ferrocyanide as an electroactive probe was enhanced according to gap distance which was controlled by surface-catalyzed chemical deposition. We found that enhanced electrochemical signal leads more sensitive signal changes according to selective interaction of target and its complementary elements on the electrode or gap area. In case of Botulinum neurotoxin, the redox signal showed a time-dependent increase due to cleavage of the immobilized peptide which blocked redox cycling. Redox cycling was also hindered by Vaccinia virus and Streptococcus Pneumoniae which were selectively immobilized in the gap area.