Tuesday, 19 November 2002

This presentation is part of : Display Presentations, Section B. Physiology, Biochemistry, Toxicology, and Molecular Biology

Molecular detection of knockdown-resistant mutations in insects

J. Marshall Clark1, Si Hyeock Lee2, Hyo Jeong Kim3, Kyong-Sup Yoon1, Jian-Rong Gao1, Steven B. Symington1, and David J. Hawthorne4. (1) University of Massachusetts, Department of Entomology, Fernald Hall, Amherst, MA, (2) Seoul National University, School of Agricultural Biotechnology, College of Agriculture and life Science, Suwon, MA, South Korea, (3) University of Massachusetts, Molecular and Cell Biology, 102G Fernald Hall, Amherst, MA, (4) University of Maryland, Department of Entomology, 4112 Plant Sciences Building, College Park, MD

Two DNA-based genotyping techniques, bi-directional PCR amplification of specific allele (bi-PASA) and single stranded conformational polymorphism (SSCP), have recently been developed and compared for the detection L1014F (insensitive sodium channel) mutation associated with DDT and permethrin resistance in 16 field populations of Colorado potato beetle, Leptinotarsa decemlineata ( Say). Bi-PASA appears to be the most accurate and rapid genotyping method, however, is strongly dependent on the quality of template genomic DNA. SSCPis more reliable but somewhat less accurate than biPASA, is less dependent on template DNA quality but requires a longer processing time. Recently, two point mutations, T929I and L932F, located in the IIS5 transmembrane segment of the voltage-sensitive sodium channel alpha subunit, have been identified and found associated with permethrin resistance (kdr-type) in the human head louse, Pediculus capitis (3). Using a serial invasive signal amplification reaction (SISAR) method, we have now simultaneously detected both mutations in spite of their close proximity.

Species 1: Coleoptera Chrysomelidae Leptinotarsa decemlineata (Colorado potato beetle)
Species 2: Phthiraptera Pediculidae Pediculus capitis (human head louse)
Keywords: resistance, permethrin

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