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Title of Journal: Parasitol Res

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Abbravation: Parasitology Research

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Springer-Verlag

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DOI

10.1007/978-0-387-72968-8_4

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1432-1955

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PCR identification of Emphasis Type="Italic"Culi

Authors: Anja Stephan PeterHenning Clausen Burkhard Bauer Stephan Steuber
Publish Date: 2009/03/24
Volume: 105, Issue: 2, Pages: 367-
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Abstract

Due to the severe outbreaks of bluetongue disease BTD in the years 2006/2007 in Germany in the absence of the main African vector Culicoides imicola a rapid and easy applicable method for identification of autochthonous Culicoides spp had to be developed Morphological identification is timeconsuming rendering impossible the identification of large numbers of midges in a short period of time A polymerase chain reaction PCRbased procedure in connection with a speciesspecific primer greatly simplifies the identification process The region of internal transcribed spacer 1 ITS1 of the ribosomal DNA has shown great potential for developing a reliable PCRbased procedure Culicoides midges were caught with ultravioletlight traps installed on different farms in Germany during 2007 and 2008 The midges were mounted on slides and morphologically characterised Midge DNA was extracted and the ITS1 region amplified using conservative primers Potential primer regions within ITS1 were determined and a speciesspecific Culicoides dewulfi primer was developed to correctly identify autochthonous C dewulfi one of the suspected BTV vectors in northwestern Europe The developed primer was used to identify C dewulfi in a pool of Culicoides midges from a farm in the state of Brandenburg

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