Establishment of Loop-mediated Isothermal Amplification Assay Detection Method of Equine Herpesviruses Type 1 |
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| Authors: | YANG Yi-ping LIU Jian-hua RAN Duo-liang XU Quan-yuan LI Na Jiaerken |
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| Institution: | College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China |
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| Abstract: | The objective of this study was to establish a rapid detection method of loop-mediated isothermal amplification assay (LAMP) for equine herpesvirus-1 (EHV-1) and evaluate the reliability of the method. Designing several pairs of LAMP primers targeting to the conserved sequence of gBgene of EHV-1, using the LAMP Real Time Turbidimeter LA-320 meter to monitor the reaction so as to screen the primers and optimize the conditions, we established a LAMP method which could amplify specific DNA of EHV-1, then added SYBR Green Ⅰ to judge the result by eye. Specific DNA of EHV-1 was amplified efficient under 65 ℃ in 50 min. There was no cross reaction to other similar viruses such as EHV-4, and the reaction system also had very high sensitivity, 10-4 dilution multiple target could be detected, the established LAMP was 10 times higher than ordinary PCR. After the reaction, add SYBR Green Ⅰ to observe results, the results were consistent with LAMP Real Time Turbidimeter LA-320 meter. Compared LAMP method result with PCR method result of four clinical samples, coincidence rate was 100%. The LAMP method in this test was rapid, specific, sensitive, easy operation and low equipment requirement, and had application prospects. |
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| Keywords: | equine herpesvirus-1 (EHV-1) loop-mediated isothermal amplification assay (LAMP) rapid detection |
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