Development of Loop-mediated isothermal amplification method for rapid detection of Streptococcus iniae, the causative agent of streptococcicosis in fish

• Published in: Journal of basic microbiology 2012-04, Vol.52 (2), p.116-122
• Main Authors: Cai, Shuang-Hu, Wang, Bei, Lu, Yi-Shan, Jian, Ji-Chang, Wu, Zao-He
• Format: Article
• Language: English
• Subjects: Animals; Aquaculture; Base Sequence; DNA Primers - genetics; DNA, Bacterial - isolation & purification; Fish Diseases - diagnosis; Fish Diseases - microbiology; Fishes - microbiology; lctO gene; Loop-mediated isothermal amplification; Nucleic Acid Amplification Techniques - methods; Rapid detection; Sensitivity and Specificity; Streptococcus - genetics; Streptococcus - isolation & purification; Streptococcus iniae; Tilapia
• ISSN: 0233-111X; 1521-4028
• DOI: 10.1002/jobm.201100082

Streptococcus iniae is a major pathogen that causes sever economic losses in tilapia aquaculture. A set of four specific primers was designed by targeting lctO gene. With Bst DNA polymerase, the target DNA can be clearly amplified for 60 min at 64 °C in a simple water bath. The sensitivity of the LAMP assay for the detection of S. iniae is about 12.4 cells per reaction in both of pure cultures and added fish tissues cultures. LAMP products could be judged with agar gel or naked eye after addition of SYBR Green I. There were no cross‐reactions with other bacterial strains indicating high specificity of the LAMP. The LAMP method was also applied to detect S. iniae ‐infected tilapia tissues effectively. The LAMP assay reported here indicates the potential usefulness of the technique as a valuable simple, rapid alternative procedure for the detection of S. iniae during streptococcicosis monitoring of cultured fish. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)