DEVELOPMENT OF A MOLECULAR DIAGNOSTIC TECHNIQUE FOR THE DETECTION OF BACULOVIRUS INFECTING OAK TASAR SILKWORM, ANTHERAEA PROYLEI

Abstract

Antheraea proylei, a temperate oak tasar silkworm, is one of the economically important silkworms reared for the production of oak tasar silk. A. proylei is frequently infected with A. proylei nucleopolyhedrovirus (AnprNPV), which causes tiger band disease leading to severe economic loss in oak tasar silk production. Therefore, development of an accurate diagnostic tool may facilitate early detection of pathogen, thus preventing massive economic loss. In the current study, we have developed a real-time quantitative polymerase chain reaction (RT-qPCR) for diagnosis of AnprNPV. The primers specific to p94 gene of AnprNPV were designed to investigate their sensitivity and specificity. The AnprNPV detection limit in RT-qPCR was found to be 2.5 × 101 copy number with 98.12% efficiency. The developed diagnostic technique is 100 times more sensitive than the conventional PCR for the detection of AnprNPV. Further, the technique was validated with filed samples wherein the AnprNPV viral loads in the oak tasar silkworms were ranged from 103 to 1010 copies/l.