Development of Loop‐mediated Isothermal Amplification (LAMP) for the Rapid Detection of Streptococcus agalactiae in Tilapia, Oreochromis niloticus. Issue 5 (October 2014)

• Record Type: Journal Article
• Authors: Ke, Xiaoli; Huo, Huanhuan; Lu, Maixin; Liu, Zhigang; Zhu, Huaping; Gao, Fengying
• Is Part Of: Journal of the World Aquaculture Society. Volume 45:Issue 5(2014)
• Page Start: 586
• Page End: 594
• Publication Date: 2014-10
• Subjects: Aquaculture -- Periodicals; 639.8
• DOI: 10.1111/jwas.12145
• Languages: English
• ISSNs: 0893-8849

Abstract

<abstract abstract-type="main" id="jwas12145-abs-0001"> <title> <bold>Abstract</bold> </title> <p id="jwas12145-para-0001"> <italic>Streptococcus agalactiae</italic> is one of the major causative agents of tilapia streptococcosis, which has caused severe economic losses in aquaculture. Rapid and accurate detection of the pathogen is necessary to limit losses because of this disease. In this study, a loop‐mediated isothermal amplification (LAMP) assay was developed for the detection of <italic>S. agalactiae</italic>. Firstly, a set of four primers was designed to target the <italic>cfb</italic> gene of <italic>S. agalactiae</italic>. Then, using <italic>Bst</italic> DNA polymerase, the LAMP assay was performed at 65 C for 60 min and terminated at 80 C for 10 min in a simple water bath. Positive or negative results could be observed by direct visual inspection. There were no cross reactions with other bacterial species, indicating high specificity of the LAMP assay. The sensitivity of the LAMP assay for detecting <italic>S</italic>. <italic>agalactiae</italic> was about 20 cells per reaction. Moreover, the developed closed‐tube step has greatly improved the LAMP system. The LAMP method was also applied to detect <italic>S. agalactiae</italic> in infected tilapia tissue, demonstrating usefulness in diagnostics. Overall, this study showed that the <italic>cfb</italic>‐based LAMP assay was an effective method to detect <italic>S. agalactiae</italic> rapidly.</p> </abstract>