The Experts below are selected from a list of 6168 Experts worldwide ranked by ideXlab platform
I Bright S Singh - One of the best experts on this subject based on the ideXlab platform.
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primary hemocyte culture of penaeus monodon as an in vitro model for white spot syndrome Virus Titration viral and immune related gene expression and cytotoxicity assays
Journal of Invertebrate Pathology, 2010Co-Authors: Seena Jose, A Mohandas, Rosamma Philip, I Bright S SinghAbstract:Abstract Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome Virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1×), tryptose phosphate broth (2.95 g l−1), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 μg ml−1 chloramphenicol, 100 μg ml−1 streptomycin and 100 IU ml−1 penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-2′-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining Virus titer as MTT50/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC50. The primary hemocyte culture could serve as a model for WSSV Titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals.
Amir H. Noormohammadi - One of the best experts on this subject based on the ideXlab platform.
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development of a sybr green quantitative polymerase chain reaction assay for rapid detection and quantification of infectious laryngotracheitis Virus
Avian Pathology, 2011Co-Authors: Alireza Mahmoudian, Mauricio J C Coppo, Naomi C Kirkpatrick, Joanne M. Devlin, Glenn F Browning, Philip F. Markham, Amir H. NoormohammadiAbstract:Infectious laryngotracheitis is an acute viral respiratory disease of chickens with a worldwide distribution. Sensitive detection of the causative herpesVirus is particularly important because it can persist in the host at a very low copy number and be transmitted to other birds. Quantification of viral genome copy number is also useful for clinical investigations and experimental studies. In the study presented here, a quantitative polymerase chain reaction (qPCR) assay was developed using SYBR Green chemistry and the viral gene UL15a to detect and quantify infectious laryngotracheitis Virus (ILTV) in ILTV-inoculated chicken embryos or naturally infected birds. The specificity of the assay was confirmed using a panel of viral and bacterial pathogens of poultry. The sensitivity of the assay was compared with two conventional PCR assays, Virus Titration and an antigen-detecting enzyme-linked immunosorbent assay. The qPCR developed in this study was highly sensitive and specific, and has potential for quant...
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development of a sybr green quantitative polymerase chain reaction assay for rapid detection and quantification of infectious laryngotracheitis Virus
Avian Pathology, 2011Co-Authors: Alireza Mahmoudian, Mauricio J C Coppo, Naomi C Kirkpatrick, Joanne M. Devlin, Glenn F Browning, Philip F. Markham, Sangwon Lee, Amir H. NoormohammadiAbstract:Infectious laryngotracheitis is an acute viral respiratory disease of chickens with a worldwide distribution. Sensitive detection of the causative herpesVirus is particularly important because it can persist in the host at a very low copy number and be transmitted to other birds. Quantification of viral genome copy number is also useful for clinical investigations and experimental studies. In the study presented here, a quantitative polymerase chain reaction (qPCR) assay was developed using SYBR Green chemistry and the viral gene UL15a to detect and quantify infectious laryngotracheitis Virus (ILTV) in ILTV-inoculated chicken embryos or naturally infected birds. The specificity of the assay was confirmed using a panel of viral and bacterial pathogens of poultry. The sensitivity of the assay was compared with two conventional PCR assays, Virus Titration and an antigen-detecting enzyme-linked immunosorbent assay. The qPCR developed in this study was highly sensitive and specific, and has potential for quantification of ILTV in tissues from naturally and experimentally infected birds and embryos.
Seena Jose - One of the best experts on this subject based on the ideXlab platform.
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primary hemocyte culture of penaeus monodon as an in vitro model for white spot syndrome Virus Titration viral and immune related gene expression and cytotoxicity assays
Journal of Invertebrate Pathology, 2010Co-Authors: Seena Jose, A Mohandas, Rosamma Philip, I Bright S SinghAbstract:Abstract Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome Virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1×), tryptose phosphate broth (2.95 g l−1), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 μg ml−1 chloramphenicol, 100 μg ml−1 streptomycin and 100 IU ml−1 penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-2′-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining Virus titer as MTT50/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC50. The primary hemocyte culture could serve as a model for WSSV Titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals.
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Primary hemocyte culture of Penaeus monodon as an in vitro model for white spot syndrome Virus Titration, viral and immune related gene expression and cytotoxicity assays
'Elsevier BV', 2010Co-Authors: Bright Singh I S, Rosamma Philip, Mohandas A, Seena JoseAbstract:Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome Virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1 ), tryptose phosphate broth (2.95 g l 1), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 lgml 1 chloramphenicol, 100 lgml 1 streptomycin and 100 IU ml 1 penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-20-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining Virus titer as MTT50/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC50. The primary hemocyte culture could serve as a model for WSSV Titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicalsCochin University of Science and TechnologyJournal of Invertebrate Pathology 105 (2010) 312–32
Angel L. Carrascosa - One of the best experts on this subject based on the ideXlab platform.
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Plaque assay for African swine fever Virus on swine macrophages
Archives of Virology, 2002Co-Authors: María J. Bustos, Marisa Nogal, Yolanda Revilla, Angel L. CarrascosaAbstract:A plaque assay developed to detect the infection of African Swine Fever Virus on swine macrophages is described. Plaques were generated by all of the Virus isolates tested. The method is suitable not only for Virus Titration but also for the selection of clones in protocols for isolation/purification of recombinant Viruses.
Ningshao Xia - One of the best experts on this subject based on the ideXlab platform.
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establishment of a rapid elispot assay for influenza Virus Titration and neutralizing antibody detection
Journal of Medical Virology, 2021Co-Authors: Guosong Wang, Pengfei Huang, Junping Hong, Ruiqi Chen, Lina Lin, Qiangyuan Han, Honglin Chen, Yixin Chen, Ningshao XiaAbstract:Seasonal influenza is an acute respiratory infection causing around 500 000 global deaths annually. There is an unmet medical need to develop more effective antiviral drugs and vaccines against influenza infection. A rapid, accurate, high-throughput Titration assay for influenza Virus particles or neutralizing antibodies would be extremely useful in these research fields. However, commonly used methods such as tissue culture infective dose and plaque-forming units (PFU) for Virus particle quantification, and the plaque reduction neutralization test (PRNT) for antibody determination are time-consuming, laborious, and have limited accuracy. In this study, we developed an efficient assay based on the enzyme-linked immunospot (ELISPOT) technique for the influenza Virus and neutralizing antibody Titration. Two broad-spectrum antibodies recognizing the nucleoproteins of influenza A and B Viruses were used in the assay to broadly and highly sensitively detect influenza Virus-infected cells at 16 hours postinfection. An optimized cell culture medium with no tosyl phenylalanyl chloromethyl ketone trypsin and high dose oseltamivir acid was used to improve quantitation accuracy. This ELISPOT assay displayed a good correlation (R2 = 0.9851) with the PFU assay when used to titrate 30 influenza Virus isolates. The assay was also applied to measure influenza-neutralizing antibodies in 40 human sera samples, showing a good correlation (R2 = 0.9965) with the PRNT assay. This ELISPOT Titration assay is a rapid, accurate, high-throughput assay for quantification of influenza Virus and neutralizing antibodies, and provides a powerful tool for research into and development of drugs and vaccines against influenza.
