1.Generation, characterization, and application in serodiagnosis of recombinant swine vesicular disease virus-like particles.
Wanhong XU ; Melissa GOOLIA ; Tim SALO ; Zhidong ZHANG ; Ming YANG
Journal of Veterinary Science 2017;18(S1):361-370
Swine vesicular disease (SVD) is a highly contagious viral disease that causes vesicular disease in pigs. The importance of the disease is due to its indistinguishable clinical signs from those of foot-and-mouth disease, which prevents international trade of swine and related products. SVD-specific antibody detection via an enzyme-linked immunosorbent assay (ELISA) is the most versatile and commonly used method for SVD surveillance and export certification. Inactivated SVD virus is the commonly used antigen in SVD-related ELISA. A recombinant SVD virus-like particle (VLP) was generated by using a Bac-to-Bac baculovirus expression system. Results of SVD-VLP analyses from electron microscopy, western blotting, immunofluorescent assay, and mass spectrometry showed that the recombinant SVD-VLP morphologically resemble authentic SVD viruses. The SVD-VLP was evaluated as a replacement for inactivated whole SVD virus in competitive and isotype-specific ELISAs for the detection of antibodies against SVD virus. The recombinant SVD-VLP assay produced results similar to those from inactivated whole virus antigen ELISA. The VLP-based ELISA results were comparable to those from the virus neutralization test for antibody detection in pigs experimentally inoculated with SVD virus. Use of the recombinant SVD-VLP is a safe and valuable alternative to using SVD virus antigen in diagnostic assays.
Animals
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Antibodies
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Baculoviridae
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Blotting, Western
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Certification
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Enterovirus B, Human
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Enzyme-Linked Immunosorbent Assay
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Foot-and-Mouth Disease
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Mass Spectrometry
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Methods
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Microscopy, Electron
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Neutralization Tests
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Serologic Tests*
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Swine Vesicular Disease*
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Swine*
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Virus Diseases
2.Establishment of RT- LAMP for rapid detection of foot-and-mouth disease virus.
Jian LI ; Qin CHEN ; Wei XIONG ; Xue-En FANG
Chinese Journal of Virology 2009;25(2):137-142
A rapid detection of foot-and-mouth disease virus (FMDV) was established by using reverse transcription loop-mediated isothermal amplification (RT-LAMP) method, meanwhile its specificity and sensitivity were assessed. The results showed that the FMDV RNA could be amplified by incubation at 65degrees C for only 1h using six primers designed based on FMDV polyprotein gene and the amplification products could be detected easily by naked-eye. There is no cross reaction with other virus such as SVDV, SFV and PPV by detecting their RNA samples. The detection limit of this method was found to be 10(-5) dilution of RNA sample which was 100-fold higher than that of PCR and 10-fold higher than real-time PCR.
Animals
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DNA Primers
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Foot-and-Mouth Disease
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prevention & control
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virology
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Foot-and-Mouth Disease Virus
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genetics
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isolation & purification
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Genome, Viral
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genetics
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Nucleic Acid Amplification Techniques
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RNA, Viral
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Reverse Transcriptase Polymerase Chain Reaction
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methods
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Sensitivity and Specificity
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Swine Vesicular Disease
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virology