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- Title
猪附红细胞体eno基因可视化LAMP检测方法的建立与应用.
- Authors
金丽兰; 付兢锋; 马浩原; 王静璇; 李长春; 薛书江; 高 旭
- Abstract
In order to establish a fast, convenient loop- mediated isothermal amplification method (LAMP) that can detect Mycoplasma suis (M. suis) at the grassroots level, this study referred to the M. suis genome (NC_015153) registered in GenBank. The α- enolase (eno) gene is the target gene and 4 specific primers were designed and synthesized based on this gene. After optimization of the reaction conditions, the LAMP method established in this study has a ratio of inner and outer primers of 16, which can using SYBR Green I at 63℃ for 50 minutes. The result is directly judged by naked eyes. This method has no crossreactivity with Toxoplasma gondii, Escherichia coli, Streptococcus suis type I, Theileria sergiomas, and Eperythrozoites of bovine origin, and has strong specificity. The sensitivity test showed that the minimum detection concentration was 30 copies/µL on M. suis eno. The detection limit was 30 copies/µL, which was 100 times more sensitive than the ordinary PCR detection method.The result of variation within and between groups was consistent, indicating that its repeatability was good. The established LAMP detection method was used for 84 porcine blood samples suspected of being infected with M. suis, and the results showed that the LAMP test results were consistent with the PCR method test results, indicating that the LAMP test established in this study could be applied to the detection of M. suis clinical samples. The LAMP method established in this study was simple, specific, sensitive and repeatable, and the results could be read by the naked eye. It was suitable for field detection and field application of M. suis infection in pig farms.
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2020, Vol 42, Issue 12, p1239
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202011015