Crabtree M B, Savage H M, Miller B R
Arbovirus Diseases Branch, National Center for Infectious Diseases, Fort Collins, Colorado, USA.
Am J Trop Med Hyg. 1995 Jul;53(1):105-9.
Culex pipiens complex mosquitoes (Cx. p. pipiens and Cx. p. quinquefasciatus) are among the principal vectors of St. Louis encephalitis (SLE) virus in the eastern United States; Cx. restuans and Cx. salinarius play secondary roles in the transmission and maintenance of the virus cycle. Accurate identification of these three species in field collections is required for epidemiologic studies of SLE virus transmission. We have developed a polymerase chain reaction (PCR) assay for this purpose. Species-specific PCR primers were designed based on interspecies nucleic acid sequence variation in the first and second internal transcribed spacers (ITS1 and ITS2) of the nuclear ribosomal DNA gene array; however, insufficient variation was detected to differentiate between subspecies of the Cx. pipiens complex. The primers were used together in a single amplification reaction to correctly identify specimens to species using genomic DNA extracted from whole individual mosquitoes, DNA from triturated mosquito pools, or crude DNA from mosquito heads or legs.
致倦库蚊复合组蚊虫（致倦库蚊和淡色库蚊）是美国东部圣路易斯脑炎（SLE）病毒的主要传播媒介之一；骚扰库蚊和盐泽库蚊在病毒循环的传播和维持中起次要作用。对SLE病毒传播进行流行病学研究需要准确鉴定野外采集的这三种蚊虫。为此我们开发了一种聚合酶链反应（PCR）检测方法。基于核糖体DNA基因阵列的第一和第二内部转录间隔区（ITS1和ITS2）种间核酸序列变异设计了种特异性PCR引物；然而，未检测到足够的变异来区分致倦库蚊复合组的亚种。这些引物在单一扩增反应中共同使用，以利用从整只蚊虫提取的基因组DNA、研磨后的蚊虫样本DNA或蚊虫头部或腿部的粗DNA正确鉴定到物种。
