DEVELOPMENT OF A SPECIES-DIAGNOSTIC POLYMERASE CHAIN-REACTION ASSAY FOR THE IDENTIFICATION OF CULEX VECTORS OF ST-LOUIS ENCEPHALITIS-VIRUS BASED ON INTERSPECIES SEQUENCE VARIATION IN RIBOSOMAL DNA SPACERS
DEVELOPMENT OF A SPECIES-DIAGNOSTIC POLYMERASE CHAIN-REACTION ASSAY FOR THE IDENTIFICATION OF CULEX VECTORS OF ST-LOUIS ENCEPHALITIS-VIRUS BASED ON INTERSPECIES SEQUENCE VARIATION IN RIBOSOMAL DNA SPACERS
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DOI：
10.4269/ajtmh.1995.53.105
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发表时间：
1995-07-01
影响因子：
3.3
通讯作者：
MILLER, BR
中科院分区：
文献类型：
--
作者：
CRABTREE, MB;SAVAGE, HM;MILLER, BR
尖音库蚊复合组（Cx. p. pipiens和Cx. p. quinquefasciatus）是美国东部圣路易斯脑炎（SLE）病毒的主要载体之一; Cx. restuans和Cx.盐藻在病毒循环的传播和维持中起次要作用。在SLE病毒传播的流行病学研究中，需要在野外采集中准确鉴定这三种病毒。我们已经开发了一种聚合酶链反应（PCR）检测为此目的。根据核核糖体DNA基因阵列的第一和第二内转录间隔区（ITS 1和ITS 2）中的种间核酸序列变异设计种特异性PCR引物;然而，检测到的变异不足以区分Cx的亚种。pipiens复合体引物在单一扩增反应中一起使用，以使用从整个个体蚊子提取的基因组DNA、来自磨碎的蚊子池的DNA或来自蚊子头或腿的粗DNA来正确地鉴定样本到物种。
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.