Penaeus monodon nucleopolyhedrovirus detection using an immunochromatographic strip test

An immunochromatographic strip test is described for detection of the polyhedrin protein of Penaeus monodon nucleopolyhedrovirus (PemoNPV). The test employs one monoclonal antibody (MAb MBV5) conjugated to colloidal gold to bind to polyhedrin protein and a 1:1:1 mixture of 3 other MAbs (MBV8, 14 and 21) to capture colloidal-gold MAb–protein complexes at a test (T) line on the nitrocellulose strip. A downstream control (C) line of goat anti-mouse immunoglobulin G (GAM) antibody is used to capture excess free colloidal-gold conjugated MBV5 to validate test performance. Heating of homogenates of PemoNPV-infected P. monodon postlarvae prepared in PBS for 30 min was necessary to maximize T line color intensity, and homogenates of infected postlarvae could still be scored as PemoNPV-positive when diluted 1:64. A strip test result was obtained within 15 min of sample application, and although about 200-fold lower than a one-step PCR test for PemoNPV, its detection sensitivity was comparable to a dot blot. Due to its simplicity not reliant on sophisticated equipment or specialized skills, the strip test could be adopted to screen easily for PemoNPV infections at shrimp hatcheries and farms.

► A strip test for Penaeus monodon nucleopolyhedrovirus was developed using four monoclonal antibodies.
► The detection sensitivity of the strip test was comparable to that of dot blot test and ∼200-fold less sensitive than one-step PCR.
► The result can be observed within 15 min.
► It does not require sophisticated equipment or specialized skills.