Effective health management in shrimp hatcheries: use of loop-mediated isothermal amplification (LAMP) for the rapid detection of Philippine isolates of Penaeus monodon-type baculovirus (MBV
Journal Title: Animal Biology & Animal Husbandry - International Journal of the Bioflux Society - Year 2011, Vol 3, Issue 1
Abstract
Effective health management in hatcheries is a crucial step towards the prevention of disease outbreaks during the early life stages of the shrimp. Early and sensitive detection of the different pathogens ensure that proper measures are undertaken before massive mortality occurs. In this study,the loop-mediated isothermal amplification (LAMP) assay was evaluated for the detection of the Penaeus monodon-type baculovirus (MBV), a viral pathogen that is common in shrimp hatcheries. The optimized time and temperature conditions for LAMP were 60 min and 63[sup]o[/sup]C, respectively. The assay was highly specific for MBV and did not amplify other shrimp viruses including white spot syndrome virus, infectious hypodermal and hematopoietic necrosis virus (IHHNV) and hepatopancreatic parvovirus (HPV). The sensitivity threshold of LAMP was 10 pg of DNA ml[sup]-1[/sup]or 10 fg of the genomic DNA per LAMP reaction and was 10 times more sensitive than conventional PCR in detecting MBV from infected shrimp postlarvae. As a possible application for field use, both LAMP and conventional PCR were able to detect the virus from tissues of infected shrimp postlarvae. These results demonstrate that LAMP could be used as a simple and sensitive method for the rapid field diagnostic tool for early detection of Philippine isolates of MBV in shrimp hatcheries.
Authors and Affiliations
Christopher Caipang, May Sibonga, Jane Geduspan, Mary Apines-Amar, Adelaida Calpe
Keywords
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