• Title/Summary/Keyword: Virus Isolation

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Detection of foot-and-mouth disease virus (FMDV) and avian influenza virus (AIV) from animal carcass disposal sites using real-time RT-PCR

  • Miguel, Michelle;Kim, Seon-Ho;Lee, Sang-Suk;Cho, Yong-Il
    • Korean Journal of Veterinary Service
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    • v.43 no.2
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    • pp.107-112
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    • 2020
  • Foot-and-mouth disease (FMD) and avian influenza (AI) are highly pathogenic viral disease which affects the livestock industry worldwide. Outbreak of these viruses causes great impact in the livestock industry; thus, disease infected animals were immediately disposed. Burial is the commonly used disposal method for deceased animals. However, there is potential for secondary environmental contamination, as well as the risk that infectious agents persisting in the environment due to the limited environmental controls in livestock burial sites during the decomposition of the carcasses. Therefore, this study aimed to investigate the detection of FMD and AI viruses from animal carcass disposal sites using real-time reverse transcription PCR. Soil samples of more than three years post-burial from livestock carcass disposal sites were collected and processed RNA isolation using a commercial extraction kit. The isolated RNA of the samples was used for the detection of FMDV and AIV using qRT-PCR. Based on the qPCR assay result, no viral particle was detected in the soil samples collected from the animal disposal sites. This indicates that 3 years of burial and their carcass disposal method is efficient for the control or at least reduction of spread infections in the surrounding environment.

Isolation of Defense-Related Genes from Nicotiana glutinosa Infected by Tobacco Mosaic Virus Using a Modified Differential Screening

  • Park, Kyung-Soon;Suh, Mi-Chung;Cheong, Jong-Joo;Park, Doil
    • The Plant Pathology Journal
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    • v.15 no.5
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    • pp.295-301
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    • 1999
  • Many of plant defense responses are consequence of transcriptional activation of related genes. We have developed a modified differential screening procedure to isolate tobacco genes that are involved in the defense responses against TMV infection. A cDNA library was constructed from Nicotiana glutinosa leaves infected by TMV under temperature shift conditions. Each of plasmid DNA in the library was hybridized on a set of slot blots to a pool of cDNA probes prepared from either TMV-infected or mock-treated tobacco leaves. Among 900 plasmid DNAs, 81 clones exhibiting significantly enhanced or reduced level of hybridization to either probe were selected for nucleotide sequencing. The clones were listed into 61 genes considering redundancy between the sequences. The genes were identified to be defense-related genes including PR-genes and genes involved in primary or secondary metabolisms. This results supports the implication that plant defense process entails a major shift in total cellular metabolisms rather than activation of a limited number of defense-related genes. Expression patterns of a number of defense-related genes. Expression patterns of a number of selected genes were examined in northern blot analyses. It is notable that the clone 630 of unknown function exhibits expression pattern similar to those of previously known PR-genes. Experiments to elucidate the roles in defense mechanism of a couple of genes newly identified in this study are in progress.

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Isolation and molecular characterization of feline panleukopenia viruses from Korean cats

  • Yang, Dong-Kun;Park, Yu-Ri;Park, Yeseul;An, Sungjun;Choi, Sung-Suk;Park, Jungwon;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.62 no.1
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    • pp.10.1-10.9
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    • 2022
  • Feline panleukopenia virus (FPV) causes fatal leukopenia and severe hemorrhagic diarrhea in cats. Although FPV isolates have been reported worldwide from several animals, the biological and genetic features of South Korean FPVs remain unclear. We characterized molecularly South Korean FPV isolates. Crandell-Rees feline kidney (CRFK) cells were used to isolate FPV from 60 organ homogenates. The isolates were confirmed to be FPVs via analyses of cytopathic effects, immunofluorescence studies, electron microscopy, and polymerase chain reaction. Viral genetic analyses used the full VP2 sequences. Eight isolates propagated in CRFK cells were confirmed to be FPVs. All isolates yielded viral titers ranging from 104.5 to 106.0 TCID50/mL 5 days after inoculation into CRFK cells and exhibited hemagglutination titers ranging from 27 to 212 (using pig erythrocytes). The Korean FPV isolates grew well in cat cells such as CRFK and Fcwf-4 cells. The FPV isolates were most similar to the KS42 strain isolated from a Korean cat in 2008. The FPV isolates will serve as useful antigens in future sero-epidemiological studies and will aid in the development of diagnostic tools.

Biological and Molecular Characterization of a Korean Isolate of Orthotospovirus chrysanthinecrocaulis (Formerly Chrysanthemum Stem Necrosis Virus) Isolated from Chrysanthemum morifolium

  • Seong Hyeon Yoon;Su Bin Lee;Eseul Baek;Ho-Jong Ju;Ju-Yeon Yoon
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.286-294
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    • 2023
  • Biological and molecular characterization of a Korean isolate of Orthotospovirus chrysanthinecrocaulis (formerly known as chrysanthemum stem necrosis virus, CSNV) isolated from Chrysanthemum morifolium was determined using host range and sequence analysis in this study. Twenty-three species of indicator plants inoculated mechanically CSNV-Kr was investigated for determination of host range. CSNV-Kr induced various local and systemic symptoms in the inoculated plant species. CSNV-Kr could not infect three plant species and induced symptomless in systemic leaves in Nicotiana tabacum cultivars, though the plant samples reacted positively with the antiserum to CSNV by double-antibody sandwich-enzyme-linked immunosorbent assay. The complete genome sequence of CSNV-Kr was determined. The L RNA of CSNV-Kr consists of 8,959 nucleotides (nt) and encodes a putative RNA-dependent RNA polymerase. The M RNA of CSNV-Kr consists of 4,835 nt and encodes the movement protein (NSm) and the glycoprotein precursor (Gn/Gc protein). The S RNA of CNSV-Kr consists of 2,836 nt and encodes NSs protein and N protein. The Gn/Gc and N sequence of CSNV-Kr were compared with those of previously published CSNV isolates originating from different countries at nucleotide and amino acid levels. The Gn/GC sequence of CSNV-Kr shared 98.8-99.5% identity with CSNV isolated from other countries and the N sequence of CSNV-Kr shared 98.8-99.6% identity. No particular region of variability could be found in either grouping of viruses. All of the CSNV isolates did not show any relationship according to geographical origins and isolation hosts, suggesting no distinct segregation of the CSNV isolates.

Epidemiology and Clincal Analysis of Acute Viral Respiratory Tract Infections in Children(September, 1998-May, 2003) (소아 급성 바이러스성 하기도 감염의 유행 및 임상양상 (1998년 9월-2003년 5월))

  • Lee, Su-Jin;Shin, Eon-Woo;Park, Eun-Young;Oh, Pil-Soo;Kim, Kwang-Nam;Yoon, Hae-Sun;Lee, Kyu-Man
    • Clinical and Experimental Pediatrics
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    • v.48 no.3
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    • pp.266-275
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    • 2005
  • Purpose : Acute respiratory tract infections are the most common illnesses in children. The great majority of these infections involving lower respiratory tracts infections(LRTIs) are caused by respiratory viruses such as respiratory syncytial virus(RSV), parainfluenza virus(PIV), influenza virus (Flu), and adenovirus(ADV), etc. Our purpose was to determine seasonal epidemiology and clinical characteristic features of each viral infection. Methods : Nasopharyngeal aspirate(NPA)s were collected from 4,554 hospitalized children diagnosed as LRTIs on the first day of admission. The study period was from September 1998(Autumn) through May 2003(Spring). Respiratory viruses were detected in 881(19 percent) cases by isolation of the virus or by antigen detection method using indirect immunofluorescent staining. We reviewed the medical records of 837 cases retrospectively. Results : The identified pathogens were RSV in 485 cases(55 percent), PIV in 152 cases(17 percent), FluA in 114 cases(13 percent), ADV in 79 cases(9 percent) and FluB in 51 cases(6 percent). Outbreaks of RSV occurred every year, mostly in the November through December period and of PIV in the April through June period. LRTIs by FluA reached the highest level in January, 2002. FluB infection showed an outbreak in April, 2002. The clinical diagnoses of viral LRTIs were bronchiolitis in 395 cases(47 percent), pneumonia in 305 cases(36 percent), croup in 73 cases(9 percent) and tracheobronchitis in 64 cases(8 percent). Conclusion : Viruses are one of the major etiologic agents of acute LRTIs in chidren. Therefore, we must continue to study their seasonal occurrence and clinical features to focus on management, and also for reasons of prevention.

Acute Respiratory Infections in the Neonatal Intensive Care Unit: Isolation of Viruses and the Characteristics of Respiratory Syncytial Virus Infection (신생아집중치료실에서의 급성 호흡기 감염-바이러스의 검출 및 respiratory syncytial virus 감염의 임상적 특징)

  • Park, Sun-Hee;Cho, Hye-Jung;Shim, So-Yeon;Son, Dong-Woo;Eun, Byung-Wook;Sun, Yong-Han;Tchah, Hann;Jeon, In-Sang
    • Neonatal Medicine
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    • v.16 no.2
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    • pp.182-189
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    • 2009
  • Purpose: Recently, it is easy to find the causal virus of acute respiratory infections using multiplex RT-PCR. The aim of this study is to show the distribution of respiratory viruses and to define the characteristics of respiratory syncytial virus (RSV) infections compared to other respiratory viral infections. Methods: This was a prospective observational study conducted in the NICU. The infants with acute respiratory infections were performed multiplex RT-PCR using nasal swabs. The demographics, initial symptoms, course of illness, and laboratory and imaging findings were recorded. The infants were divided into RSV and No RSV groups. Results: Twenty-three infants (50%) were in the RSV group. Rhinovirus was the second most common virus. Coinfections with two viruses accounted for 6.5% of respiratory infections. The number of preterm infants, exposure to cigarette smoke and having siblings were not different between the two groups. Infections in the postnatal care center were more common in the RSV group than the No RSV group (60.9% vs. 21.7%, P=.007). Dyspnea (34.8% vs. 8.7%, P=.032) and pneumonia (73.9% vs. 43.5%, P=.036) were more common in the RSV group. The RSV group frequently needed oxygen (52.5% vs. 13.0%, P=.005) and received nothing by mouth (43.5% vs. 13.0%, P=.022). The incidence of right upper consolidation was higher in RSV group (56.5% vs. 8.7%, P=.001). Conclusion: This study showed that other viruses than RSV can induce respiratory infections in neonates and young infants born prematurely. RSV infections have a more severe course of illness than other respiratory viruses. We have to be careful of prevention even for healthy neonates especially in crowed situations, such as the postnatal care center.

Investigation of atrophic piglets diseases in northern area of the Gyeongnam province, Korea (경남 북부지역 위축자돈에 대한 질병조사)

  • Kim, Hyeong-Su;Seong, Min-Ho;Han, Kwon-Seek;Park, Jung-Yong;Shin, Yoo-Gyeong;Jeong, Myeong-Ho;Park, Dong-Yeop;Koh, Phil-Ok
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.1-7
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    • 2015
  • This study was investigated to diagnose pathogenic organisms of atrophic piglets in northern area of the Gyeongnam province, Korea. Samples such as feces, blood and necropsy specimens of 42 atrophic piglets (${\leq}10$ weeks old) were taken from May to December 2013 for this survey. Samples were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and bacteria isolation for detection of pathogenic agents. 93 pathogens were isolated from 42 samples can be classified into ll groups. We identified bacterial agents in 56 cases (60.2%) and viral agents in 31 cases (33.3%). However, 6 cases (6.5%) were undetected. Among these pathogens, the most prevalent disease were porcine reproductive and respiratory syndrome (PRRS) in 22 cases (23.7%). The major diseases were Colibacillosis in 15 cases (16.1%), Glasser's disease in 12 cases (12.9%), and porcine epidemic disease (PED) in 9 cases (9.7%). Mixed infections were accounted for 77.8% of atrophic piglets. In particular, the rate of mixed infections with PRRS virus showed the highest frequency (71.4%). In addition, there is a seasonal variation. Viral pathogens were dominantly detected in winter, but in the rest of the season bacterial agents were mainly detected. Gastrointestinal diseases occurred mainly in the pre-weaning piglets, the respiratory diseases and wasting diseases occurred mainly in the post-weaning piglets.

Isolation and characterization of cultured chicken oviduct epithelial cells and in vitro validation of constructed ovalbumin promoter in these cells

  • Yang, Hyeon;Lee, Bo Ram;Lee, Hwi-Cheul;Jung, Sun Keun;Kim, Ji-Youn;No, Jingu;Shanmugam, Sureshkumar;Jo, Yong Jin;Lee, Haesun;Hwang, Seongsoo;Byun, Sung June
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1321-1330
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    • 2021
  • Objective: Transgenic hens hold a great promise to produce various valuable proteins. Through virus transduction into stage X embryo, the transgene expression under the control of constructed chicken ovalbumin promoters has been successfully achieved. However, a validation system that can evaluate differently developed ovalbumin promoters in in vitro, remains to be developed. Methods: In the present study, chicken oviduct epithelial cells (cOECs) were isolated from oviduct tissue and shortly cultured with keratinocyte complete medium supplemented with chicken serum. The isolated cells were characterized with immunofluorescence, western blot, and flow cytometry using oviduct-specific marker. Chicken mutated ovalbumin promoter (Mut-4.4-kb-pOV) was validated in these cells using luciferase reporter analysis. Results: The isolated cOECs revealed that the oviduct-specific marker, ovalbumin protein, was clearly detected by immunofluorescence, western blot, and flow cytometry analysis revealed that approximately 79.40% of the cells contained this protein. Also, luciferase reporter analysis showed that the constructed Mut-4.4-kb-pOV exhibited 7.1-fold (p<0.001) higher activity in the cOECs. Conclusion: Collectively, these results demonstrate the efficient isolation and characterization of cOECs and validate the activity of the constructed ovalbumin promoter in the cultured cOECs. The in vitro validation of the recombinant promoter activity in cOECs can facilitate the production of efficient transgenic chickens for potential use as bioreactors.

Surviving Covid-19 Diagnosis Among Registered Nurses: Reactions, Consequences, and Coping Mechanisms

  • Gladys Mbuthia;Doris Machaki;Sheila Shaibu;Rachel W. Kimani
    • Safety and Health at Work
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    • v.14 no.4
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    • pp.467-475
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    • 2023
  • Background: To mitigate the spread of Covid-19, nurses infected with the virus were required to isolate themselves from their families and community. Isolated patients were reported to have experienced mental distress, posttraumatic stress disorder symptoms, and suicide. Though studies have reported the psychological impact of the Covid-19 pandemic, less is known about the lived experiences of nurses who survived Covid-19 infection in sub-Saharan Africa. Methods: A descriptive phenomenological approach was used to study the lived experiences of registered nurses who survived Covid-19 disease. In-depth interviews were conducted among nurses diagnosed with Covid-19 from two hospitals in Kenya between March and May, 2021. Purposive and snowball sampling were used to recruit registered nurses. Data were analyzed using Giorgi's steps of analysis. Results: The study included ten nurses between 29 and 45 years of age. Nurses' experiences encompassed three themes: diagnosis reaction, consequences, and coping. Reactions to the diagnosis included fear, anxiety, and sadness. The consequence of the diagnosis and isolation was stigma, isolation, and loneliness. Nurses coping mechanisms included acceptance, creating routines, support, and spirituality. Conclusion: Our findings aid in understanding how nurses experienced Covid-19 infection as patients and will provide evidence-based content for supporting nurses in future pandemics. Moreover, as we acknowledge the heroic contribution of frontline healthcare workers during the Covid-19 pandemic, it is prudent to recognize the considerable occupational risk as they balance their duty to care, and the risk of infection to themselves and their families.

Clinical Analysis of Influenza in Children and Rapid Antigen Detection Test on First Half of the Year 2004 in Busan (2004 상반기 부산 지역에서 유행한 인플루엔자의 임상 역학적 분석 및 인플루엔자 진단에 있어서의 신속 항원 검사법)

  • Choi, So Young;Lee, Na Young;Kim, Sung Mi;Kim, Gil Heun;Jung, Jin Hwa;Choi, Im Jung;Cho, Kyung Soon
    • Pediatric Infection and Vaccine
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    • v.11 no.2
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    • pp.158-169
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    • 2004
  • Purpose : Although influenza is one of the most important cause of acute respiratory tract infections in children, virus isolation is not popular and there are only a few clinical studies on influenza and diagnostic methods. We evaluated the epidemiological and clinical features of influenza in children and rapid antigen detection test(QuickVue influenza test) on fist half of the year 2004 in Busan. Methods : From January 2004 to June 2004, throat swab and nasal secretion were obtained and cultured for the isolation of influenza virus and tested by rapid antigen detection test(QuickVue influenza test) in children with suspected influenza infections. The medical records of patients with influenza virus infection were reviewed retrospectively. Results : Influenza viruses were isolated in 79(17.2%) out of 621 patients examined. Influenza virus was isolated mainly from March to April 2004. The ratio of male and female with influenza virus infection was 1.2 : 1 with median age of 4 years 6month. The most common clinical diagnosis of influenza virus infection was bronchitis. There was no difference between influenza A and B infection in clinical diagnosis and symptoms. All patients recovered without severe complication. The sensitivity obtained for rapid antigen detection test (QuickVue influenza test) was 93.6% and the specificity was 80.2%, the positive predictive value 40.8%, the negative predictive value 98.8%. Conclusion : With rapid antigen detection test, it is possible early detection of influenza in children. reduction in use of antimicrobial agent and early use of antiviral agent.

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