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Production of recombinant nucleocapsid protein of Newcastle disease virus in Escherichia coli for a diagnostic ELISA
2009
Kim, H.I., Institute of Cheilbio, Ansan, Republic of Korea | Park, K.P., Institute of Cheilbio, Ansan, Republic of Korea | Park, C.H., Institute of Cheilbio, Ansan, Republic of Korea | Cho, H.A., Institute of Cheilbio, Ansan, Republic of Korea | Yang, H.S., Institute of Cheilbio, Ansan, Republic of Korea | Hahn, T.W., Kangwon National University, Chuncheon, Republic of Korea
Transmission of avian viruses both bird-to-bird and from birds to non-avian species is a major health concern. Newcastle disease virus (NDV) is an economically important avian virus that poses substantial risks to the poultry industry. Rapid and sensitive diagnostic methods, such as the enzyme-linked immunosorbent assay (ELISA), are required to track such infections. To develop an ELISA for detecting anti-NDV antibody in avian sera, the nucleocapsid protein (NCP) gene of the NDV La Sota strain was cloned and expressed in Escherichia coli and the 513-amino acid recombinant NCP was purified by Ni-NTA affinity chromatography. To evaluate its ability to replace NDV whole virus antigen as a coating antigen, NCP-coated and whole NDV-coated ELISAs were tested and compared using a panel of NDV positive antisera from chickens. Results using purified NCP were highly correlated with those obtained using whole NDV (r=0.927), demonstrating that recombinant NCP expressed in Escherichia coli is a suitable substitute antigen for whole NDV in a diagnostic ELISA.
Показать больше [+] Меньше [-]Agreement of two ELISAs for Mycobacterium avium subspecies paratuberculosis in cattle in Korea
2009
Lee, K.W., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Jung, B.Y., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Hwang, I.Y., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Lee, S.H., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Kim, J.Y., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Kim, Y.H., Gyeongbuk Veterinary Service Laboratory, Daegu, Republic of Korea | Lee, S.H., Institute of Livestock and Veterinary Research, Jeonju, Republic of Korea | Moon, O.K., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Lee, O.S., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea
Paratuberculosis caused by Mycobacterium avium subspecies paratuberculosis (Mpt) is a chronic infectious enteric disease with deleterious impact on the performance in ruminants. In Korea, ELISA has been introduced to detect antibodies to Mpt in individual cattle. However, comparison study with ELISA has not been studied until now. In total, a panel of 899 serum samples obtained from dairy cattle was analyzed with two commercial ELISAs for Mpt to assess the performance. Two ELISAs employed in this study were both licensed worldwide. Two ELISAs applied onto same serum samples showed the morderate agreement (kappa value = 0.06). There was non-significant McNemar test (p=0.0614) between two ELISA results indicating that each proportion detected by two kits did not differ. In addition, the percent agreement between two ELISA results was turned out to be 96.8% which interpreted excellent reproducibility. It was shown from this study that two ELISAs revealed moderate kappa agreement performance. The implication raised is that when ELISAs as diagnostics are used to detect Mpt in individual cattle, positive reaction by either ELISA should be interpreted as serologically Mpt positive due to presumed low sensitivity of ELISAs and their test agreement being less than 100%.
Показать больше [+] Меньше [-]Immunological relationships of FMD vaccine strain and Asia1 field isolate from East Asia
2009
Park, J.H., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Ko, Y.J., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Kim, S.M., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Lee, H.S., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Lee, K.N., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea | Cho, I.S., National Veterinary Research and Quarantine Service, Anyang, Republic of Korea
Foot-and-mouth disease (FMD) is the most contagious disease of mammals. The use of inactivated vaccine can be chosen to prevent or control FMD. However, vaccination against one serotype of FMDV doses not cross-protect against other serotypes and may not protect fully against some strains of the same serotype. Appropriate selection of vaccine strain is an important element in the control of FMD. The immunity of vaccine antigens should be matched against newly circulating viruses. The phylogenetic analysis of serotype Asia1 reported from China, Mongolia, North Korea and Russia since 2005 shows that they are all classified into genetic group V, but the strain, Asia1/Shamir (ISR/89) which have been used as a vaccine strain in Korea, is clustered into different genetic group. So, in this study the serological relationship between the isolate (Asia1/MOG/05; MOG) and the Shamir stain was determined by ELISA and virus neutralization test. Even though the matching value of the virus (MOG) against the vaccinated sera in target animals was not so high, the vaccinated animals elicited antibodies enough for protection after vaccinated once or twice. Conclusively, we suggest that the vaccine containing Asia1/Shamir antigen could protect the genetic group V strains circulating in East Asia currently if vaccinated twice or the more.
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