• Title/Summary/Keyword: Virus detection

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Real-Time PCR for Validation of Minute Virus of Mice Safety during the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Minute Virus of Mice 안전성 검증을 위한 Real-Time PCR)

  • Lee, Dong-Hyuck;Cho, Hang-Mee;Kim, Hyun-Mi;Lee, Jung-Suk;Kim, In-Seop
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.12-20
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to minute virus of mice(MVM), and there are several reports of MVM contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the MVM safety, a real-time PCR method was developed for quantitative detection of MVM in cell lines, raw materials, manufacturing processes, and final products as well as MVM clearance validation. Specific primers for amplification of MVM DNA was selected, and MVM DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $6{\times}10^{-2}TCID_{50}/mL$. The real-time PCR method was proven to be reproducible and very specific to MVM. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with MVM. MVM DNA could be Quantified in CHO cell as well as culture supernatant. When the real-time PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of MVM.

Detection of foot-and-mouth disease virus and coxsakievirus in the soil and leachate of modeled carcass burial site (시험 가축 매몰지 토양 및 침출수 내에서의 구제역 바이러스 검출)

  • Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.255-261
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    • 2012
  • Foot and mouth disease (FMD) is highly infectious disease of cloven-hoofed animals, particularly cattle, sheep, pigs and goats. Last outbreak reported in November, 2010 induced the enormous social and economical impacts. Culling of infected animals, movement control, and vaccination are the major control measures of FMD. The aim of this study was to detection foot-and-mouth disease virus (FMDV) in the soil and leachate from modeling burial for pig carcass as measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). FMDV and Coxsakievirus B1 (CVB1) were detected in soil by week 16 and Coxsakievirus B1 (CVB1) by weeks 12, respectively. FMDV and CVB1 also detected by weeks 8 in the leachate. Results from this study provides an evidence that FMDV could be inactivated for safe of pig carcasses infected with FMDV within 4 month in the carcass burial site.

PCR-Based Detection of Densovirus Infection in Silkworm (Bombyx mori L.)

  • Hou Chengxiang;Li Muwang;Gui Zhongzheng;Xu Anying;Guo Xijie
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.2
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    • pp.135-138
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    • 2005
  • Two pairs of DNA primers were designed for the detection of the Zhenjiang (China) strain of Bombyx mori densonucleosis virus (BmDNV-Z). These primers were designed from the nucleotide sequence of major structural protein gene (putative VD1-ORF2). PCR amplification was attempted from different issues (including silk gland, blood, skin and midgut) and feces of the silkworm which infected wit BmDNV-Z were amplified by PCR. Both of the primers gave expected size of in the DNA bands from midgut and feces, but not in the DNA of silk gland, blood and skin. The two bands were sequenced, and their sequence were same as the sequence designed for. BmDNV-Z could be successfully detected in single silkworm after it was infected for 12 hrs, and could not be detected before 9 hrs after infected.

Porcine circovirus: detection of antibodies and virus antigen in Chungbdk area (Porcine circovirus에 대한 항체가 조사 및 바이러스 항원 확인)

  • 강신석;박재명;이종진;류재윤;최해연
    • Korean Journal of Veterinary Service
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    • v.24 no.2
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    • pp.127-132
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    • 2001
  • Porcine circoviruses(PCV) are the smallest nonenveloped DNA viruses containing a unique single-stranded circular genome. No recognized link was found between PCV infection of pig and disease. But the PCV consistently identified from postweaning multisystemic wasting syndrome(PMWS) and researches indicate that there are strong relationships between PCV and PMWS. Clinical signs were emaciation, dyspnea, high fever with normal appetite. Necropsy findings showed respiratory disease complex lesion and lymph node anomalities. An indirect-immunofluorescent antibody procedure was used to assay swine sera for the presence of PCV atibodies. Antibodies against PCV were found in an average of 20% of the samples tested. The PCV DNA was amplified from lymph nodes collected from pigs. PCV specific primers were successfully amplified PCV DNAs. Further studies are needed to determine the possible role this virus might have in disease.

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Immunohistochemistry for the Detection of Swine hepatitis E virus in the liver

  • Ha, Seung-Kwon;Chae, Chan-hee
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.28-28
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    • 2003
  • Hepatitis E virus (HEV), previously referred to as enterically transmitted non-A, non-B hepatitis, is responsible for sporadic infections as well as large epidemics of acute viral hepatitis in developing countries. The disease generally affects young adults and reportedly has a mortality rate of up to 20% in infected pregnant women. HEV was once considered to be a member of the family Caliciviridae, but the unique genomic organization of HEV has led to the removal of HEV from the family and it was provisionally classified in an unassigned family of HEV-like viruses. In situ hybridization provides any cellular detail and histological architecture.[1] However, use of in situ hybridization is largely restricted to the laboratories because this technique is the greater technical complexity and expense compared with immunohistochemistry. Therefore, the objective of this study is to develop the immunohistochemistry for the detection of swine HEV from formalin-fixed, paraffin-embedded hepatic tissues. (omitted)

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Correlation of Axillary Artery Pressure and Phase of Esophageal Impedance in Chickens

  • Nakajima, Isao;Kuwahira, Ichiro;Hori, Shuho;Mitsuhashi, Kokuryo
    • Journal of Multimedia Information System
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    • v.9 no.2
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    • pp.161-170
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    • 2022
  • Under General anesthesia with isoflurane, we insert a chicken's esophageal catheter into the near the left atrium. 1MHz radio wave was added to electrocardiogram electrodes of the esophagus, and the change of impedance (phase) was obtained by amplitude synchronous detection technique. At the same time, a thin tube is surgically inserted into the axillary artery to continuously measure blood pressure. The correlation between impedance (phase) and blood pressure was obtained. Both showed a very high correlation (R2=0.9665). It was also observed the waveform flowing from the left atrium into the left ventricle. When an individual infected with the avian influenza virus develops, the cytokine storms lead to hypotension earlier than the test for antigen-antibody reaction. In order to detect this, in the future, this impedance technique will be useful for screening individuals infected with avian influenza virus by measuring the blood pressure of chickens in cages in a non-contact manner using microwaves.

Novel pan-lineage VP1 specific degenerate primers for precise genetic characterization of serotype O foot and mouth disease virus circulating in India

  • Sagar Ashok Khulape;Jitendra Kumar Biswal;Chandrakanta Jana;Saravanan Subramaniam;Rabindra Prasad Singh
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.40.1-40.6
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    • 2023
  • Analysis of the VP1 gene sequence of the foot and mouth disease virus (FMDV) is critical to understanding viral evolution and disease epidemiology. A standard set of primers have been used for the detection and sequence analysis of the VP1 gene of FMDV directly from suspected clinical samples with limited success. The study validated VP1-specific degenerate primer-based reverse transcription polymerase chain reaction (RT-PCR) for the qualitative detection and sequencing of serotype O FMDV lineages circulating in India. The novel degenerate primer-based RT-PCR amplifying the VP1 gene can circumvent the genetic heterogeneity observed in viruses after cell culture adaptation and facilitate precise viral gene sequence analysis from clinical samples.

Infection of Mycovirus in Imported Lentinula edodes (해외도입 표고버섯의 진균바이러스 감염)

  • Lee, Song Hee;Kwak, Seo-Young;Ko, Han Kyu;Lee, Hyun-Sook
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.64-68
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    • 2014
  • Up to date several mycoviruses including Lentinula edodes Spherical Virus (LeSV) have been reported. As fungal virus was spreaded by infested hypae and spores it could be important to use virus-free spawns to eradicate the mushroom virus disease in the culture farm. We tested the imported spawns of Lentinula edodes by PCR whether LeSV was infested them or not. The primer set targeting the RdRp gene of LeSV was prepared based on partial sequence of the LeSV genome. The RT-PCR analysis showed that 87 among 88 imported spawns of L. edodes were infested by LeSV.

Application of Epstein-Barr Virus Cell Lines (CCL85 EB-3) in Performing the EBER mRNA In Situ Hybridization as a Positive Control (Epstein-Barr 바이러스 인사이투 보결합 시행시 양성대조표지로서의 버키트 림프종 세포주 (CCL85 EB-3)의 응용)

  • Kim, Sung-Sook;Han, Woon-Sup;Suh, Joo-Young;Huh, Joo-Ryung
    • The Korean Journal of Cytopathology
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    • v.7 no.1
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    • pp.38-43
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    • 1996
  • Epstein-Barr virus(EBV) is associated with a wide spectrum of benign and malignant disorders including leukoplakia, Hodgkln's lymphoma, central nervous system lymphoma, peripheral T cell lymphoma and nasopharyngeal undifferentiated carcinoma. There are several distinctive aspects of biology of the virus that are important in investigation of virus in clinical specimens. The abundant expression of the EBER mRNA transcripts makes possible the sensitive detection of latent expression in EBV-associated tumors. Although there has been a dramatic increased interest in the direct characterization of EBV in clinical specimens, there have been few studios about the effective and reliable positive controls in performing in situ hybridization technique for EBV, especially on paraffin-em bedded tissue. We applied Burkitts lymphoma ceil line as positive control in EBV in situ hydridization using Oncor Kit. The cell block of Burkitt lymphoma cell line(CCL85 EB-3) showed strong and specific positivity for EBER in situ in nuclei of EBV infected cells.

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Aerodynamic Approaches for Estimation of Waste Disease Spread in Pig Farm through Airborne Contaminants (양돈장의 소모성질병 확산 분석을 위한 현장 모니터링 및 공기유동학적 분석)

  • Seo, Il-Hwan;Lee, In-Bok;Moon, Oun-Kyung;Kwon, Kyeong-Seok
    • Journal of The Korean Society of Agricultural Engineers
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    • v.56 no.1
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    • pp.41-49
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    • 2014
  • Pig chronic wasting disease, including porcine reproductive and respiratory syndrome (PRRS) and postweaning multisystemic wasting syndrome (PMWS), have made a continuous economic damage in pig farms. Airborne spread of livestock viruses are an important spread factor which is difficult to analyze due to invisible airflow and limitation of measurement. The objective of this study is to analyze airborne disease spread between buildings in the experimental pig farm by means of field experiment and computational fluid dynamics (CFD). The field experiments were conducted to capture airborne virus using air sampler and teflon filter along multi points in the experimental pig farm. The samples were tested in terms of virus detection resulting in positive reaction for PRRS and PCV-2 viruses, which can be a firm evidence of airborne virus spread. The CFD simulation model was developed by considering complex topography, wind conditions, building arrangement, and ventilation systems and was used to analyze airborne virus spread according to different wind conditions. The CFD computed result showed a possibility of airborne virus spread via livestock aerosol from infected pig house to neighboring pig houses according to wind directions. The CFD simulation technique is expected to provide significant data for estimating and making a counterplan against airborne disease spread.