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.
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