An improved method for PCR-base detection of nuclear polyhedrosis virus in Bombyx mori
1995
Noguchi, Y. (Saitama-ken. Sericultural Experiment Station, Kumagaya (Japan)) | Kobayashi, M. | Shimada, T.
We have already established a diagnostic technique based on the polymerase chain reaction to detect nuclear polyhedrosis virus in a silkworm population using samples containing the wastes and feces as well as larval bodies. In the present study, we utilized potassium acetate precipitation instead of phenol/chloroform treatment at the deproteinization step of DNA extraction, thus avoiding danger of applying a large amount of organic solvents. Also HCl treatment of DNA followed by ethanol precipitation was found to be effective to increase the diagnostic sensitivity. By using this improved method, we could detect a single polyhedrosis-infected larva in a 0.5, 1 and 3 kg sample for the first, second and third instar, respectively
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