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首页> 外文期刊>Applied and Environmental Microbiology >DNA hybridization assay for detection of gypsy moth nuclear polyhedrosis virus in infected gypsy moth (Lymantria dispar L.) larvae.
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DNA hybridization assay for detection of gypsy moth nuclear polyhedrosis virus in infected gypsy moth (Lymantria dispar L.) larvae.

机译：DNA杂交测定法，用于检测吉普赛蛾（Lymantria dispar L.）幼虫中的吉普赛核多角体病毒。

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Radiolabeled Lymantria dispar nuclear polyhedrosis virus DNA probes were used in a DNA hybridization assay to detect the presence of viral DNA in extracts from infected larvae. Total DNA was extracted from larvae, bound to nitrocellulose filters, and assayed for the presence of viral DNA by two methods: slot-blot vacuum filtration and whole-larval squashes. To test the assays, neonate larvae were fed droplets containing a known concentration of L. dispar nuclear polyhedrosis virus and observed for up to 10 days to determine the percentage of infected larvae. The average percent mortalities were 88.0, 60.7, 26.0, and 5.3% for larvae fed droplets containing 4.0 x 10(4), 1.0 x 10(4), 2.5 x 10(3), and 6.25 x 10(2) polyhedral inclusion bodies (PIBs) per ml, respectively. Other larvae treated with the same virus concentrations were frozen at 2, 4, and 6 days postinoculation and examined by the hybridization techniques. The average percentage of slot blots containing viral DNA equaled 81.0, 58.0, 18.0, and 6.0% for larvae blotted 4 days after treatment with 4.0 x 10(4), 1.0 x 10(4), 2.5 x 10(3), and 6.25 x 10(2) PIBs per ml, respectively, and 89.9, 52.1, 26.6, and 6.0%, respectively at 6 days postinoculation. Thus, the hybridization results were closely correlated with mortality observed in reared larvae. Hybridization of squashes of larvae frozen 4 days after receiving the above virus treatments also produced accurate measures of the incidence of virus infection.
机译：放射性标记的Lymantria dispar核多角体病毒DNA探针用于DNA杂交测定中，以检测感染的幼虫提取物中病毒DNA的存在。从幼虫中提取总DNA，结合到硝酸纤维素滤膜上，并通过两种方法检测病毒DNA的存在：狭缝印迹真空过滤和全幼南瓜。为了测试试验，给新生幼虫喂入含有已知浓度的L. dispar核多角体病毒的小滴，并观察长达10天以确定感染幼虫的百分比。含有4.0 x 10（4），1.0 x 10（4），2.5 x 10（3）和6.25 x 10（2）多面体包涵体的幼虫饲喂小滴的平均死亡率为88.0、60.7、26.0和5.3％ （PIBs）每毫升。用相同病毒浓度处理的其他幼虫在接种后第2、4和6天冷冻，并通过杂交技术进行检查。分别用4.0 x 10（4），1.0 x 10（4），2.5 x 10（3）和6.25处理后的4天印迹的幼虫，含有病毒DNA的缝隙印迹的平均百分比分别为81.0、58.0、18.0和6.0％接种后第6天，每毫升x 10（2）PIB分别为89.9、52.1、26.6和6.0％。因此，杂交结果与在饲养幼虫中观察到的死亡率密切相关。在接受上述病毒处理后4天冷冻的幼虫南瓜的杂交也产生了准确的病毒感染发生率的量度。

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