Publication: Shewanella putrefaciens in cultured tilapia detected by a new calcein-loop-mediated isothermal amplification (Ca-LAMP) method
Issued Date
2015-12-09
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ISSN
16161580
01775103
01775103
DOI
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2-s2.0-84950137942
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Mahidol University
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SCOPUS
Bibliographic Citation
Diseases of Aquatic Organisms. Vol.117, No.2 (2015), 133-143
Suggested Citation
Rungkarn Suebsing, Jantana Kampeera, Sarawut Sirithammajak, Padmaja Jayaprasad Pradeep, Sarocha Jitrakorn, Narong Arunrut, Pakkakul Sangsuriya, Vanvimon Saksmerprome, Saengchan Senapin, Boonsirm Withyachumnarnkul, Wansika Kiatpathomchai Shewanella putrefaciens in cultured tilapia detected by a new calcein-loop-mediated isothermal amplification (Ca-LAMP) method. Diseases of Aquatic Organisms. Vol.117, No.2 (2015), 133-143. doi:10.3354/dao02942 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/35052
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Title
Shewanella putrefaciens in cultured tilapia detected by a new calcein-loop-mediated isothermal amplification (Ca-LAMP) method
Abstract
© Inter-Research 2015. Shewanella putrefaciens is being increasingly isolated from a wide variety of sources and is pathogenic to many marine and freshwater fish. For better control of this pathogen, there is a need for the development of simple and inexpensive but highly specific, sensitive, and rapid detection methods suitable for application in field laboratories. Our colorogenic loop-mediated isothermal amplification (LAMP) assay combined with calcein (Ca-LAMP) for unaided visual confirmation of LAMP amplicons is a simple method for fish pathogen detection in cultured tilapia. Here, we describe the detection of S. putrefaciens using the same platform. As before, the method gave positive results (orange to green color change) in 45 min at 63°C with sensitivity 100 times higher than that of a conventional PCR assay, with no cross-amplification of other known fish bacterial pathogens tested. Using the assay with 389 samples of gonads, fertilized eggs, and fry of farmed Nile and red tilapia Oreochromis spp., 35% of samples were positive for S. putrefaciens. The highest prevalence was found in samples of gonads (55%) and fertilized eggs (55%) from adult breeding stocks, indicating that S. putrefaciens could be passed on easily to fry used for stocking production ponds. Tissue tropism assays revealed that the spleen showed the highest colonization by S. putrefaciens in naturally infected tilapia and that it would be the most suitable organ for screening and monitoring fish stocks for presence of the bacteria.
