2.5. Sensitivity and specificity testing
To determine the sensitivity of the PCR method
using primer pair (261F/R) for MBV DNA detection,
10-fold serial dilutions of purified plasmid
WS-1 containing the 261 PCR product were used
as DNA templates for PCR assay. The concentration
of plasmid DNA ranged from 1010 to 1 copy per
reaction. In addition, we tested by PCR the published
protocols (OIE, 2003) for MBV using
infected tissue.
To check for the specificity of the primer pair
(261F/R), DNA from penaeid shrimp infected with
four different penaeid shrimp viruses was used. The
extracted DNA containing baculovirus penaei (BP),
infectious hypodermal and hematopoietic necrosis
virus (IHHNV), white spot syndrome virus
(WSSV) or hepatopancreatic parvovirus (HPV) was
used as templates for PCR amplification.