Detection of white spot syndrome virus by polymerase chain reaction performed under insulated isothermal conditions

Aiming to develop a rapid, low-cost, and user-friendly system for the diagnosis of white spot syndrome virus (WSSV), a PCR assay performed in capillary tubes under insulated isothermal conditions (iiPCR assay) was established on the basis of Rayleigh–Benard convection. WSSV amplicons were generated reproducibly within 30 min from a target sequence-containing plasmid in an iiPCR device, in which a special polycarbonate capillary tube (R-tube™) was heated isothermally by a copper ring attached to its bottom and shielded by a thermal baffle around its upper half. Furthermore, WSSV-specific amplicons were produced from nucleic acid extracts of WSSV-infected Penaeus vannamei in the WSSV iiPCR assay, with sensitivity comparable to that of an OIE-certified commercial nested PCR kit (IQ2000™ WSSV Detection and Prevention System). Specificity of the WSSV iiPCR assay was demonstrated as no amplicons were generated from shrimp genomic DNA, and IHHNV, MBV, and HPV DNA. iiPCR has a potential as a low-cost method for sensitive, specific and rapid detection of pathogens.

► We established a simple and sensitive assay for WSSV on the basis of iiPCR, a convective PCR.
► WSSV iiPCR was able to generate detectable amounts of amplicons in 30 min.
► Sensitivity of the WSSV iiPCR assay was comparable to that of a nested PCR-based assay.
► The WSSV iiPCR assay generated amplicons only from WSSV-, but not from IHHNV-, MBV-, and HPV-positive samples.
► iiPCR has a potential as a low-cost method for sensitive, specific and rapid detection of pathogens.