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首页> 外文期刊>Journal of Economic Entomology >A Multiplex Real-Time PCR Assay for Screening Gypsy Moths (Lepidoptera: Erebidae) in the United States for Evidence of an Asian Genotype
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A Multiplex Real-Time PCR Assay for Screening Gypsy Moths (Lepidoptera: Erebidae) in the United States for Evidence of an Asian Genotype

机译：多重实时PCR检测筛查吉普赛蛾（鳞翅目：孔雀科）在亚洲的基因型证据

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摘要

European gypsy moth populations (Lymantria dispar L.) are well established and a proven destructive force in hardwood trees throughout the United States and Canada. Introduction of the exotic Asian gypsy moth into North America would be even more impactful, as Asian gypsy moth populations have wider host ranges, and are capable of naturally dispersing more rapidly due to female flight ability. To support early detection and exclusion of Asian gypsy moth, the U.S. Department of Agriculture (USDA) uses molecular techniques to screen moths trapped in North America for evidence of common Asian genotype. In order to strengthen U.S. domestic capacity to screen moths quickly and efficiently, we report a real-time PCR assay for this pest. A probe system using TaqMan 5' nuclease chemistry is reported for detection of an allele associated with common Asian gypsy moth genotypes. The targeted allele is located at the nuclear FS1 locus currently used by the USDA in conventional PCR tests to screen for evidence of Asian gypsy moth introductions or introgression. The diagnostic probe is successfully multiplexed with a conserved 18S probe system to detect reaction failure due to poor sample quality or quantity. The specificity, sensitivity, and repeatability of the FS1-18S multiplex real-time PCR assay were tested on laboratory-reared and field-collected moths to demonstrate diagnostic utility. Implications of the new assay as a screening tool for evidence of Asian gypsy moth introgression and introduction are discussed.
机译：欧洲的吉普赛蛾种群（Lymantria dispar L.）是公认的，并且在整个美国和加拿大的硬木树中具有公认的破坏力。由于亚洲吉普赛蛾种群具有更广泛的寄主范围，并且由于雌性飞行能力而能够更快地自然扩散，因此将奇异的亚洲吉普赛蛾引入北美的影响甚至更大。为了支持及早发现和排除亚洲吉普赛蛾，美国农业部（USDA）使用分子技术来筛选在北美被困的蛾，以寻找常见的亚洲基因型。为了增强美国国内快速有效地筛查飞蛾的能力，我们报告了该害虫的实时PCR检测方法。据报道使用TaqMan 5'核酸酶化学的探针系统可检测与常见的亚洲吉普赛蛾基因型相关的等位基因。靶向的等位基因位于美国农业部目前在常规PCR测试中使用的FS1核基因座，以筛选出引入亚洲吉普赛蛾或基因渗入的证据。诊断探针可与保守的18S探针系统成功复用，以检测由于样品质量或数量不佳而导致的反应失败。在实验室饲养的飞蛾和田间采集的飞蛾上测试了FS1-18S多重实时PCR分析的特异性，敏感性和可重复性，以证明其诊断用途。讨论了新方法作为亚洲吉普赛蛾渗入和引入证据的筛选工具的意义。

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