• Title/Summary/Keyword: 역전사 중합효소 연쇄반응

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Development and Assessment of Specific and High Sensitivity Reverse Transcription Nested Polymerase Chain Reaction Method for the Detection of Aichivirus A Monitoring in Groundwater (지하수 중 Aichivirus A 모니터링을 위한 특이적 및 고감도 이중 역전사 중합효소연쇄반응 검출법 개발 및 평가)

  • Bae, Kyung Seon;Kim, Jin-Ho;Lee, Siwon;Lee, Jin-Young;You, Kyung-A
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.190-198
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    • 2021
  • Human Aichivirus (Aichivirus A; AiV-A) is a positive-sense single-strand RNA non-enveloped virus that has been detected worldwide in various water environments including sewage, river, surface, and ground over the past decade. To develop a method with excellent sensitivity and specificity for AiV-A diagnosis from water environments such as groundwater, a combination capable of reverse transcription (RT)-nested polymerase chain reaction (PCR) was developed based on existing reported and newly designed primers. A selective method was applied to evaluate domestic drinking groundwater samples. Thus, a procedure was devised to select and subsequently identify RT-nested PCR primer sets that can successfully detect and identify AiV-A from groundwater samples. The findings will contribute to developing a better monitoring system to detect AiV-A contamination in water environments such as groundwater.

Detection of Poliovirus in Water by Cell Culture and PCR Methods (세포배양법과 PCR 방법에 의한 물에서의 폴리오 바이러스 검출)

  • 조연희;이찬희
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.198-204
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    • 2002
  • Poliovirus is a member of enterovirus which causes paralytic poliomyelitis, encephalitis and aseptic meningitis. Since poliovirus is spread by the fecal-oral route and poliovirus-contaminated water could be a potential threat for public health, detection of poliovirus in drinking water resource is important. Infectious poliovirus and poliovirus inactivated by heat or UV were used to test three detection methods such as cell culture method, reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture (ICC)-PCR. Infectious poliovirus was detected by all three methods and ICC-PCR was the most sensitive and fast in detecting poliovirus. Inactivated polioviruses could not be detected by cell culture or ICC-PCR methods. On the other hand, heat- inactivated viruses could be detected by RT-PCR. Thus it is suggested that ICC-PCR method is the most sensitive and effective in detecting infectious polioviruses in water sample.

Rapid Detection and Identification of Cucumber Mosaic Virus by Reverse Transcription and Polymerase Chain Reaction (RT-PCR) and Restriction Analysis (역전사 중합효소련쇄반응(RT-PCR)과 제한효소 분석을 이용한 오이 모자이크 바이러스의 신속한 검정과 동정)

  • Park, Won Mok
    • Journal of Plant Biology
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    • v.38 no.3
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    • pp.267-274
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    • 1995
  • Based upon the nucleotide sequence of As strain of cucumber mosaic virus (CMV-As0 RNA4, coat protein (CP) gene was selected for the design of oligonucleotide primers of polymerase chain reaction (PCR) for detection and identification of the virus. Reverse transcription and polymerase chain reaction (RT-PCR) was performed with a set of 18-mer CMV CP-specific primers to amplify a 671 bp fragment from crude nucleic acid extracts of virus-infected leaf tissues as well as purified viral RNAs. The minimum concentrations of template viral RNA and crude nucleic acids from infected tobacco tissue required to detect the virus were 1.0 fg and 1:65,536 (w/v), respectively. No PCR product was obtained when potato virus Y-VN RNA or extracts of healthy plants were used as templates in RT-PCR using the same primers. The RT-PCR detected CMV-Y strain as well as CMV-As strain. Restriction analysis of the two individual PCR amplified DNA fragments from CMV-As and CMV-Y strains showed distinct polymorphic patterns. PCR product from CMV-As has a single recognition site for EcoRI and EcoRV, respectively, and the product from CMV-Y has no site for EcoRI or EcoRV but only one site for HindIII. The RT-PCR was able to detect the virus in the tissues of infected pepper, tomato and Chinese cabbage plants.

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Characterization and RT-PCR Detection of Turnip Mosaic Virus Isolated from Chinese Cabbage in Korea (배추에서 분리한 순무 모자이크 바이러스의 특성 및 역전사 중합효소 연쇄반응법(RT-PCR)을 이용한 검정)

  • 박원목;최설란;김수중;최승국;류기현
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.223-228
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    • 1998
  • Turnip mosaic virus)TuMV-Ca) was isolated from a Chinese cabbage showing severe mosaic and black necrotic spots symptoms in Korea. The virus was identified as a strain of TuMV by its host range test, particle morphology, serology, double stranded RNA analysis. For detection of the virus, reverse transcription and polymerase chain reaction(RT-PCR) was performed with a set of 18-mer TuMV-specific primers to amplify a 876 bp DNA fragment The virus was rapidly detected from total nucleic acids of virus infected tissues as well as native viral RNA of purified virion particles by RT-PCR. Detection limit of the viral RNA by RT-PCR was 10 fg.

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Molecular Detection of Korean-type Bovine Immunodeficiency Virus by Polymerase Chain Reaction (DNA 중합효소 연쇄반응을 이용한 한국형 젖소 면역 결핍 바이러스의 검출)

  • 권오식
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.101-107
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    • 1999
  • Bovine immunodeficiency virus (BIV) which was grouped into the Lentivirinae of family Retroviridae, was known to be causing many immunodeficiency syndromes among cows. The BIV was studied worldwide during last several years for its importance in cattle industries but nothing was reported in Korea until now Thus we initially tried to study the existence of BIV in cattle around the Daegu·Kyungpook area by PCR related molecular techniques. As a prerequisite investigation for detecting Korean-type BIV, we had focused our aim into BLV infected cows because the BLV infected cows tend to show BIV infection with 5% ranges. Hence we randomly sampled fresh bloods from 248 cows and bulls near the Daegu·Kyungpook area and collected peripheral blood monocytes (PBMC) from the sample bloods. After extracting genomic DNA from the PBMC, we subjected it to PCR and Soluthern blot analysis for BIV/BLV detection. Overall, 66.9% (81/121) of the cow PBMC samples turned out to be BLV positive by PCR and the result was reconfirmed by Southern blot analysis. The value was two times higher than the previously reported results of BLV infection in Korea. The significant difference was mainly due to 1) applying highly specific methods for BLV detection such as PCR 2) that BLV was continuously spreaded in the Daegu Kyungpook area without any notice during last ten years. We also tested the BLV positive samples with the same techniques for BIV detection. And we found some BIV positives among the lot 3C samples by PCR, which had showed 100% BLV positive.

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Role of Chest Radiographs and CT Scans and the Application of Artificial Intelligence in Coronavirus Disease 2019 (코로나바이러스감염증 2019에서 흉부X선사진 및 CT의 역할과 인공지능의 적용)

  • Seung-Jin Yoo;Jin Mo Goo;Soon Ho Yoon
    • Journal of the Korean Society of Radiology
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    • v.81 no.6
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    • pp.1334-1347
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    • 2020
  • Coronavirus disease (COVID-19) has threatened public health as a global pandemic. Chest CT and radiography are crucial in managing COVID-19 in addition to reverse transcription-polymerase chain reaction, which is the gold standard for COVID-19 diagnosis. This is a review of the current status of the use of chest CT and radiography in COVID-19 diagnosis and management and anㄷ introduction of early representative studies on the application of artificial intelligence to chest CT and radiography. The authors also share their experiences to provide insights into the future value of artificial intelligence.

Cause of enteroviral infection in children in chungnam area summer, 2005 (2005년 하절기에 충남지역 소아에서의 장바이러스 감염원인)

  • Jeon, Se Yun;Choi, Suk Joo;Kim, Yong Bae;Nam, Hae Seon;Park, Kwi Sung;Baek, Kyung Ah;Park, Joon Soo
    • Clinical and Experimental Pediatrics
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    • v.49 no.11
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    • pp.1186-1193
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    • 2006
  • Purpose : Enterovirus infection is a type of viral infection that occurs relatively frequently in children during summer. It has clinical symptoms of non-specific fever, aseptic encephalomeningitis, gastrointestinal diseases, skin rash and, hand-foot-mouth disease. However, it can also occcaisionally, result in fatal symptoms like myocarditis, epicardial inflammation, transverse myelitis, quadriplegia and etc. There have been epidemic enterovirus studies, but not in the Chungnam area. Therefore, we undertook this study in order to comprehend the cause viruses in this area. Methods: We enlisted 157 children hospitalized with enteroviral infections at Soonchunhyang University hospital in Cheonan between May and August 2005. Cerebrospinal fluids or feces were collected during the acute phase after hospitalization, and observed the cytopathic effects caused by enterovirus and using reverse transcription polymerase chain reaction (RT-PCR). Results : The number of children hospitalized due to possible enteroviral infection during the period of study was 157. The number of children who tested positive with the reverse transcription polymerase chain reaction totalled 32 cases (20.4 percent). Among the children with entroviral diseases, 20 were male and 12 were female, thus the sex ratio of male to female was 1.67:1. Their clinical symptoms included fever most frequently (93.7 percent), was followed by headaches (90.0 percent), meningeal irritation signs (65.0 percent), and abdominal pain (30.0 percent). As for the type of isolated enterovirus, there were 17 cases of echovirus 18 and 6 cases of coxsackievirus B5. Furthermore, there were 2 cases of echovirus 9, 1 case of coxsackievirus A6 and coxsackievirus B3, respectively. But 5 cases were not determined by genotype. Conclusion : Echovirus 18 is circulating in Korea. We reported on identified enteroviruses, including echovirus 18, using RT-PCR in the Chungnam area during the summer of 2005.

Multiple Genetic Marker Analysis with Using Quantitative RT-PCR in Gastric Cancer (위암에서 정량적 역전사 중합효소연쇄반응을 이용한 다중 표지자 분석)

  • Yoo, Moon-Won;Lee, Hyuk-Joon;Choi, Soo-Min;Yu, Ji-Eun;Hur, Keun;Kim, Young-Kook;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.7 no.2
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    • pp.59-66
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    • 2007
  • Purpose: This study was aimed at evaluating the diagnostic validity of peritoneal dissemination of gastric cancer cells by performing multiple genetic marker analysis via quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in gastric cancer cell lines and gastric cancer tissues. Materials and Methods: Quantitative RT-PCR was performed on 12 human gastric cancer cell lines and 10 gastric cancer tissues with four mRNAs of carcinoembryonic antigen (CEA), Cytokeratin 20 (CK20), dopa decarboxylase (DDC), and L-3-phosphoserine phosphatase (L3PP). Results: Out of the 12 human gastric cancer cell lines we tested, CEA was overexpressed in four cell lines (33%), CK20 in one (8%), DDC in six (50%) and L3PP was expessed in all the lines (100%). Out of the 10 gastric cancer tissues we tested, CEA was overexpressed in nine tissues, CK20 in eight, DOC in nine and L3PP was overexpressed in all the tissues. L3PP was overexpressed in all the gastric cancer cell lines and tissues, but the levels of overexpression were lower than those of CEA and DDC. Conclusion: Multiple genetic marker analysis can compensate for the weak points of single marker analysis when testing gastric cancer, and three mRNAs of CEA, DDC and L3PP can be used as candidate genes.

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Analysis of Antigenic and Genetic Variability of G-protein of Respiratory Syncytial Virus Subgroup A Isolated in Korea over 8 Years(1990~1998) (국내에서 분리된 Respiratory Syncytial Virus A 아군의 항원성의 변이와 G-단백 mRNA의 RT-PCR 생산물의 제한효소 처리 및 염기 서열 결정을 통한 유전자 변이의 분석)

  • Choi, Eun Hwa;Park, Ki Ho;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.6 no.2
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    • pp.219-233
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    • 1999
  • Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

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Study on PPARγ-dependent adipogenesis regulation by testosterone using transient transfection assays (일시적인 형질전환 분석을 이용한 testosterone에 의한 PPARγ-의존적 지방세포생성 조절에 관한 연구)

  • Jeong, Sunhyo
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.2
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    • pp.482-487
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    • 2014
  • This study is to investigate the effects of testosterone on adipogenesis and its molecular mechanism using RT-PCR analysis and transient transfection assays. Castrated(CAST) mice treated with testosterone had lower white adipose tissue weights and expression of adipocyte-specific genes($PPAR{\gamma}$ and aP2) than CAST control mice. Consistent with the in vivo data, testosterone treatment inhibited triglyceride accumulation and expression of adipocyte-specific genes($PPAR{\gamma}$ and aP2) in differentiated 3T3-L1 cells compared with control group. Testosterone-activated androgen receptor(AR) repressed the luciferase reporter gene activity induced by $PPAR{\gamma}$ transfection. Thus, these results suggest that testosterone downregulates the actions of $PPAR{\gamma}$ on adipogenesis through AR.