• Title/Summary/Keyword: Virus diagnosis

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Development and Characterization of Monoclonal Antibodies against Nucleoprotein for Diagnosis of Influenza A Virus

  • Nguyen, Hong Phuong;Kwak, Chaewon;Heo, Chang-Kyu;Cho, Eun Wie;Yang, Jihyun;Poo, Haryoung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.809-815
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    • 2018
  • Influenza, which is a highly contagious disease caused by the influenza A virus, continues to be a major health concern worldwide. Although the accurate and early diagnosis of influenza virus infection is important for controlling the spread of this disease and rapidly initiating antiviral therapy, the current influenza diagnostic kits are limited by their low sensitivity. In this study, we developed several new influenza nucleoprotein (NP)-specific monoclonal antibodies (mAbs) and compared their sensitivity and specificity of those with commercially available anti-NP mAbs. Three mAbs, designated M24.11, M34.3, and M34.33, exhibited higher reactivities to recombinant NPs and A/Puerto Rico/8/1934 (H1N1) viral lysates compared with the commercial mAbs, as assessed using enzyme-linked immunosorbent assays. M34.3 and M34.33 showed higher reactivities with A/California/04/09 (pandemic H1N1) and A/Philippines/2/82 (H3N2) viral lysates than the commercial mAbs. In contrast, M24.11 had marked reactivity with H3N2 but not with pandemic H1N1. Immunofluorescent confocal microscopy showed that the three mAbs effectively detected the presence of influenza virus in lung tissues of mice infected with A/Puerto Rico/8/1934. These results indicate that the newly developed M34.3 and M34.33 mAbs could be useful for the development of influenza diagnostics.

BK virus-associated hemorrhagic cystitis after pediatric stem cell transplantation

  • Han, Seung Beom;Cho, Bin;Kang, Jin Han
    • Clinical and Experimental Pediatrics
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    • v.57 no.12
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    • pp.514-519
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    • 2014
  • Hemorrhagic cystitis is a common stem cell transplantation-related complication. The incidence of early-onset hemorrhagic cystitis, which is related to the pretransplant conditioning regimen, has decreased with the concomitant use of mesna and hyperhydration. However, late-onset hemorrhagic cystitis, which is usually caused by the BK virus, continues to develop. Although the BK virus is the most common pathogenic microorganism of poststem cell transplantation late-onset hemorrhagic cystitis, pediatricians outside the hemato-oncology and nephrology specialties tend to be unfamiliar with hemorrhagic cystitis and the BK virus. Moreover, no standard guidelines for the early diagnosis and treatment of BK virus-associated hemorrhagic cystitis after stem cell transplantation have been established. Here, we briefly introduce poststem cell transplantation BK virus-associated hemorrhagic cystitis.

A Case of Epstein Barr Virus-Associated Hemophagocytic Syndrome Confirmed by mRNA In Situ Hybridization and Polymerase Chain Reaction (mRNA In Situ Hybridization으로 확인된 Epstein Barr Virus-Associated Hemophagocytic Syndrome 1례)

  • Kim, Chung Han;Yang, Chang Hyun;Sohn, Young Mo;Kim, Hoguen
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.200-206
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    • 1996
  • Virus associated hemophagocytic syndrome(VAHS), a class II histiocytosis syndrome, is characterized by high fever, liver dysfunction, coagulation abnormalities, and generalized histiocytic proliferation with marked hemophagocytosis in bone marrow and lymph nodes. VAHS is associated with several viral infections including Epstein Barr virus which has a relatively high mortality rate. We report a fatal case of Epstein Barr virus associated hemophagocytic syndrome and its diagnosis by mRNA in situ hybridization and polymerase chain reaction. A brief review of related literaure is also presented.

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The production of monoclonal antibodies against canine distemper virus (Canine Distemper Virus에 대한 단클론성 항체의 생산)

  • Kim, Tae-jong;Kim, Se-young
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.375-381
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    • 1997
  • The purpose of the production of monoclonal antibodies aganist the Canine distemper virus(CDV) were perfect diagnosis and a new approach to treat canine distemper because the diagnosis and treatment of canine distemper were difficult. Canine distemper virus(CDV) was purified using saturated ammonium sulfate, and injected into hind footpads of BALB/c mouse. 12-15 days later, popliteal lymph node(PN) cells were harvested and fused with SP2/O myeloma cells. Characteristics of monoclonal antibodies were analysed. 1. 9 hybridomas produce the specific antibody against CDV. 2. 6 monoclonal antibodies are against intranuclear and cytoplasmic component of CDV, and 3 monoclonal antibodies are against cytoplasmic inclusions. 3. All monoclonal antibodies did not react with other 5 different viruses (CAV-I, CAV-II, CCV, CPV and CPIV) and react with another CDV-FXNO strain. 4. 3 monoclonal antibodies have neutralizing activity against CDV. 5. Antigenic difference was observed between CDV by IFA.

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

  • Ju, Ho-Jong
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.385-389
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    • 2011
  • A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RTPCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.

Diagnosis of Potato Leafroll Virus with Digoxigenin-labeled cRNA Probes (Digoxigenin으로 표지된 cRNA 프로브를 이용한 감자잎말림바이러스(PLRV)의 짐단)

  • 서효원;함영일;오승은;신관용;최장경
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.636-641
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    • 1998
  • Digoxigenin (DIG) was used to prepare nucleic acid probe for the detection of RNA of potato leafroll virus (PLRV) in the potato leaf extracts. The 0.6 kb coat protein (CP) gene cDNA of PLRV in plasmid pSPT 18 vector was labeled with digoxigenin by in vitro run-off transcription and then used for cRNA probe. In the several buffers tested for increase the total RNA extraction efficiency AMES buffer was the most suitable for this detection method. The RNA extracts from potato leaves shown symptoms of PLRV were dot blotted onto nylon membrane and hybridized with labeled RNA probes. After hybridization, labeled RNA bound to PLRV RNA on membrane was detected with anti-digoxigenin alkaline phosphatase. 5-bromo-4-chloro-3-indolyl-phosphate/nitroblue tetrazolium (NBT) salt and CSPD were used as substrate for colorimetric and film exposure detection, respectively. These detection methods were very sensitive allowing for detection of 1/32 diluted total RNA extract from 100 mg leaf tissue.

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Seroprevalence of antiboby to porcine reproductive and respiratory syndrome virus from pig sera collected from breeding herds (번식돈에서 돼지 생식기 호흡기증 바이러스 항체 분포 조사)

  • 박최규;김현수
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.89-94
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    • 2004
  • Total 2,451 sera collected from pig farms nationwide were tested for the detection of porcine reproductive and respiratory syndrome(PRRS) virus antibodies. The results were analyzed between different geographic regions, types of breeding pigs, and different years. The overall seroprevalence of PRRS virus antibodies for 3 years was 32.4%(705/2,451). The seroprevalence of PRRS virus antibodies in years 2000, 2001, 2002, and 2004 was 33.4% (284/850), 38.6%(291/754), 33.3%(155/466), and 17.1%(65/381), respectively. The seropevalence of PRRS virus antibody in sow in years 2000, 2001, 2002 and 2003 was 31.7%, 28.4%, 29.6%, and 13.4%, respectively. The seropevalence of PRRS virus antibody in gilts in years 2000, 2001, 2002 and 2003 was 36.6%, 67.4%, 54.7%, and 33.9%, respectively. The seropevalence of PRRS virus antibody in boars in years 2000, 2001 and 2003 was 45.7%, 36.4%, and 100%, respectively. No boar serum sample was submitted for the diagnosis of PRRS virus antibody in the year 2000. High seroprevalence of the PRRS virus antibody in sow, gilts and boars indicates that the infected breeding pigs are the major source of the PRRS virus infection, and also play an important role in spreading the PRRS virus between fan mates or herds.

A Review of Detection Methods for the Plant Viruses

  • Jeong, Joo-Jin;Ju, Ho-Jong;Noh, Jaejong
    • Research in Plant Disease
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    • v.20 no.3
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    • pp.173-181
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    • 2014
  • The early and accurate detection of plant viruses is an essential component to control those. Because the globalization of trade by free trade agreement (FTA) and the rapid climate change promote the country-to-country transfer of viruses and their hosts and vectors, diagnosis of viral diseases is getting more important. Because symptoms of viral diseases are not distinct with great variety and are confused with those of abiotic stresses, symptomatic diagnosis may not be appropriate. From the last three decades, enzyme-linked immunosorbent assays (ELISAs), developed based on serological principle, have been widely used. However, ELISAs to detect plant viruses decrease due to some limitations such as availability of antibody for target virus, cost to produce antibody, requirement of large volume of sample, and time to complete ELISAs. Many advanced techniques allow overcoming demerits of ELISAs. Since the polymerase chain reaction (PCR) developed as a technique to amplify target DNA, PCR evolved to many variants with greater sensitivity than ELISAs. Many systems of plant virus detection are reviewed here, which includes immunological-based detection system, PCR techniques, and hybridization-based methods such as microarray. Some of techniques have been used in practical, while some are still under developing to get the level of confidence for actual use.

Development of an RT-PCR assay and its positive clone for plant quarantine inspection of American plum line pattern virus in Korea

  • Da-Som Lee;Junghwa Lee;Seong-Jin Lee;Seungmo Lim;Jaeyong Chun
    • Korean Journal of Agricultural Science
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    • v.49 no.4
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    • pp.821-831
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    • 2022
  • American plum line pattern virus (APLPV), a member of the genus Ilarvirus in the family Bromoviridae, is one of the plant quarantine pathogens in Korea. In this study, 15 candidate primer sets were designed and examined to develop a reverse transcription polymerase chain reaction (RT-PCR) assay for plant quarantine inspection of APLPV. Using APLPV-infected and healthy samples, the primer sets were assessed for APLPV detection. To confirm the occurrence of nonspecific reactions, six ilarviruses (Apple mosaic virus, Asparagus virus 2, Blueberry shock virus, Prune dwarf virus, Prunus necrotic ringspot virus, and Tobacco streak virus) and 10 target plants (Prunus mume, P. yedoensis, P. persica, P. armeniaca, P. dulcis, P. tomentosa, P. avium, P. glandulosa, P. salicina, and P. cerasifera) were examined. Finally, two primer sets were selected. These primer sets could generate the expected amplicons even with at least 1 ng of the total RNA template in concentration-dependent amplifications. In addition, a positive clone was developed for use as a positive control in the abovementioned RT-PCR assay.

Herpes Zoster Meningitis Confirmed by Detection of Varicella-Zoster Virus DNA Using the Polymerase Chain Reaction -A case report- (중합효소 연쇄반응을 이용한 Varicella-Zoster Virus DNA 검출로 확인된 대상포진 수막염 -증례 보고-)

  • Heo, Hu Man;Choi, Yu Sun;Park, Seong Kyu
    • The Korean Journal of Pain
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    • v.18 no.2
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    • pp.210-213
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    • 2005
  • Acute viral meningitis and myositis are rare complications of varicella-zoster virus (VZV) reactivation. A 71-years-old immunocompetent man, who presented with lower back pain radiating to the left lower extremities, developed vesicles on the L5 dermatomal area. The next day, he had complained of aberrant vesicles on the trunk, face and scalp, with generalized myalgia, headache and dizziness. He was confirmed with VZV meningitis and myositis, as demonstrated by the presence of VZV DNA in the blood and cerebral spinal fluid using a polymerase chain reaction (PCR) amplification. PCR has been used in patients with a VZV infection associated neurological symptoms, and provides a useful tool for the early diagnosis of VZV-associated neurological disease. The patient was treated with bed rest, with intravenous acyclovir for the VZV infection, and intravenous Patient-controlled Analgesia for pain management and the prevention of postherpetic neuralgia. When he visited the outpatient department 3 months later, the skin lesion, leg pain, headache and myalgia had all improved, without sequelae. Here, this case is reported, with a discussion of the relevant literature on its diagnosis and management.