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)

Effective health management in hatcheries is a crucial step towards the prevention of diseaseoutbreaks during the early life stages of the shrimp. Early and sensitive detection of the differentpathogens 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 Penaeusmonodon-type baculovirus (MBV), a viral pathogen that is common in shrimp hatcheries. The optimizedtime and temperature conditions for LAMP were 60 min and 63oC, respectively. The assay was highlyspecific for MBV and did not amplify other shrimp viruses including white spot syndrome virus, infectioushypodermal and hematopoietic necrosis virus (IHHNV) and hepatopancreatic parvovirus (HPV). Thesensitivity threshold of LAMP was 10 pg of DNA ml-1 or 10 fg of the genomic DNA per LAMP reaction andwas 10 times more sensitive than conventional PCR in detecting MBV from infected shrimp postlarvae. Asa possible application for field use, both LAMP and conventional PCR were able to detect the virus fromtissues of infected shrimp postlarvae. These results demonstrate that LAMP could be used as a simpleand sensitive method for the rapid field diagnostic tool for early detection of Philippine isolates of MBV inshrimp hatcheries.