ポリメラーゼ連鎖反応による飼育残沙を含めた大量試料からの核多角体病蚕の検出

DOI 日本農学文献記事索引 Web Site 被引用文献3件

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  • An application of the polymelase chain reaction for practical diagnosis of the nuclear polyhedrosis in large-scale culture of Bombyx mori.
  • ポリメラーゼ レンサ ハンノウ ニ ヨル シイク ザンサ オ フクメ タ タイ

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We developed a simple and highly sensitive method for the detection of the nuclear polyhedrosis virus in a large-scale culture population of Bombyx mori, using the polymelase chain reaction (PCR). Newly exuviated 3rd instar larvae were crushed in a plastic bags, mixed with 8 volumes of alkali solution (0.2 M Na2CO3, 0.2M NaCl, 20mM EDTA, pH 10.8) and incubated at 25°C for 30min to dissolve the polyhedra. The lysate was treated with detergent and Proteinase K. DNA was purified with phenol, diluted with water and used as a template for PCR. We could detect a single polyhedrosis larva in a population of 20, 000 larvae even if the sample contained mulberry leaves or rearing wastes. Because our method is much more sensitive than other conventional methods, we propose it as a tool for a practical diagnosis of the nuclear polyhedrosis in cooperative rearing.

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