Guerrero F D, Kunz S E, Kammlah D
Knipling-Bushland U.S. Livestock Insects Research Laboratory, USDA-ARS, Kerrville, TX 78028, USA.
J Med Entomol. 1998 Sep;35(5):710-5. doi: 10.1093/jmedent/35.5.710.
A polymerase chain reaction (PCR)-based rapid screening procedure was developed to test individual horn flies, Haematobia irritans irritans (L.), for the presence of a specific nucleotide substitution in the sodium channel gene sequence that has been associated with pyrethroid resistance. By a systematic optimization of reaction conditions and judicious choice of PCR primers differing in DNA sequence by a single nucleotide, we identified pyrethroid-susceptible or resistant sodium channel alleles in individual flies. Laboratory and field populations were examined by both the PCR assay and conventional filter paper bioassays with the pyrethroid cyhalothrin to verify that populations containing greater proportions of individuals with the resistant sodium channel allele DNA sequence also had higher bioassay LC50 values. The PCR assay for resistance alleles gave definitive information on the genotype of an individual fly and detected the presence of heterozygous individuals that might serve as reservoirs of resistance genes in field populations.
开发了一种基于聚合酶链反应（PCR）的快速筛选程序，用于检测个体角蝇（Haematobia irritans irritans (L.)）钠通道基因序列中与拟除虫菊酯抗性相关的特定核苷酸取代情况。通过系统优化反应条件并明智地选择DNA序列仅相差一个核苷酸的PCR引物，我们在个体苍蝇中鉴定出了对拟除虫菊酯敏感或抗性的钠通道等位基因。通过PCR检测和使用拟除虫菊酯氯氟氰菊酯的传统滤纸生物测定法对实验室和野外种群进行了检查，以验证含有更高比例具有抗性钠通道等位基因DNA序列个体的种群也具有更高的生物测定LC50值。针对抗性等位基因的PCR检测提供了关于单个苍蝇基因型的确切信息，并检测到杂合个体的存在，这些个体可能是野外种群中抗性基因的储存库。
