A Fluorogenic Quantitative PCR Assay for Valid Quantity of SeMNPV in Sample

With the software BLAST from NCBI , the unique gene ORF22 and gene ORF40 of Spodoptera exigua nuclear polyhedrovirus (SeMNPV) were selected and cloned into the pMD18-T vector. The purified pMD18-T-ORF22 and pMD18-T-ORF40 were used as standard samples, genome of quantified SeMNPV were used as control, a real-time TaqMan-based PCR assay was developed to rapidly detect valid quantity of SeMNPV. The equation of standard curves were con=10(-0.282CT+9.965) （R2=0.9997）and con=10(-0.296CT+9.945) (R2=0.9995. This study indicated that a SeMNPV polyhedrobody contained 102 nucleic molecules. SeMNPV complex as detection sample contained 633 PIBs/mg、691 PIBs/mg SeMNPV polyhedrobodies, the two methods were in accordance with each other, and was in accordance with the actual quantity in the samples.