Abstract
Timely pond-side detection of white spot syndrome virus (WSSV) plays a critical role in the implementation of bio-security measures to help minimize economic losses caused by white spot syndrome disease, an important threat to shrimp aquaculture industry worldwide. A portable device, namely POCKIT™, became available recently to complete fluorescent probe-based insulated isothermal PCR (iiPCR), and automatic data detection and interpretation within one hour. Taking advantage of this platform, the IQ Plus™ WSSV Kit with POCKIT system was established to allow simple and easy WSSV detection for on-site users. The assay was first evaluated for its analytical sensitivity and specificity performance. The 95% limit of detection (LOD) of the assay was 17 copies of WSSV genomic DNA per reaction (95% confidence interval [CI], 13 to 24 copies per reaction). The established assay has detection sensitivity similar to that of OIE-registered IQ2000™ WSSV Detection and Protection System with serial dilutions of WSSV-positive Litopenaeus vannamei DNA. No cross-reaction signals were generated from infectious hypodermal and haematopoietic necrosis virus (IHHNV), monodon baculovirus (MBV), and hepatopancreatic parvovirus (HPV) positive samples. Accuracy analysis using700 L. vannamei of known WSSV infection status shows that the established assayhassensitivity93.5% (95% CI: 90.61–95.56%) and specificity 97% (95% CI: 94.31–98.50%). Furthermore, no discrepancy was found between the two assays when 100 random L. vannamei samples were tested in parallel. Finally, excellent correlation was observed among test results of three batches of reagents with 64 samples analyzed in three different laboratories. Working in a portable device, IQ Plus™ WSSV Kit with POCKIT system allows reliable, sensitive and specific on-site detection of WSSV in L. vannamei.
Highlights
White spot syndrome, an OIE-listed disease, has resulted in severe economic losses in the shrimp aquaculture industry worldwide [1]
The limit of detection (LOD) determined by probit analysis with white spot syndrome virus (WSSV) genomic DNA was17 copies per reaction (95% confidence interval [CI], 13 to 24 copies per reaction)
The assay could detect as low as 17 copies of WSSV genomic DNA per reaction (95% LOD) and reach detection limits similar to those of the nested Polymerase chain reaction (PCR)-based IQ2000TM WSSV Detection and Protection System (DPS), an OIE-registered WSSV detection assay
Summary
An OIE-listed disease, has resulted in severe economic losses in the shrimp aquaculture industry worldwide [1]. The disease is caused by white spot syndrome virus (WSSV), an enveloped large double-stranded DNA virus belonging to genus Whispovirus in the family Nimaviridae. Juvenile and adult penaeid shrimp could all be infected by WSSV both vertically and horizontally. WSSV replicates preferentially in the cuticular epithelium and subcuticular connective tissues, as well as in the antennal gland, and haematopoietic tissues in shrimp. Factors such as rapid salinity and temperature changes can help activate WSSV in apparently healthy carriers and trigger the onset of the disease. Samples from pleopods, gills, haemolymph, stomach or abdominal muscle are recommended for diagnostic testing in shrimp [2]
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