Identification of I. ricinus, I. persulcatus and I. trianguliceps species by multiplex PCR

Kairi Värv; Anna Ivanova; Julia Geller; Jaanus Remm; Kertu Jaik; Nina Tikunova; Vera Rar; Åke Lundkvist; Irina Golovljova

doi:10.1016/j.ttbdis.2016.11.004

Identification of I. ricinus, I. persulcatus and I. trianguliceps species by multiplex PCR

Ticks Tick Borne Dis. 2017 Feb;8(2):235-240. doi: 10.1016/j.ttbdis.2016.11.004. Epub 2016 Nov 10.

Authors

Kairi Värv¹, Anna Ivanova², Julia Geller², Jaanus Remm³, Kertu Jaik³, Nina Tikunova⁴, Vera Rar⁴, Åke Lundkvist⁵, Irina Golovljova⁶

Affiliations

¹ Department of Virology, National Institute for Health Development, Tallinn, Estonia; Department of Gene Technology, Tallinn University of Technology, Tallinn, Estonia; Department of Medical Biochemistry and Microbiology, Zoonotic Science Center (ZSC), Uppsala University, Sweden. Electronic address: kairi.varv@tai.ee.
² Department of Virology, National Institute for Health Development, Tallinn, Estonia.
³ Department of Zoology, University of Tartu, Tartu, Estonia.
⁴ Laboratory of Molecular Microbiology, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.
⁵ Department of Medical Biochemistry and Microbiology, Zoonotic Science Center (ZSC), Uppsala University, Sweden.
⁶ Department of Virology, National Institute for Health Development, Tallinn, Estonia; Department of Medical Biochemistry and Microbiology, Zoonotic Science Center (ZSC), Uppsala University, Sweden; Laboratory of Molecular Microbiology, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation; University of Information Technologies, Mechanics and Optics, St. Petersburg, Russian Federation.

PMID: 27856176
DOI: 10.1016/j.ttbdis.2016.11.004

Abstract

Correct identification of tick species is an essential requirement for any scientific study engaged in tick-associated research. However, morphological identification can lead to misinterpretations, especially when dealing with vector-host research and sub-adult, engorged or damaged specimens. To overcome this limitation, we developed a novel assay to discriminate between Ixodes ricinus, I. persulcatus and I. trianguliceps species collected from rodents or vegetation, using the second internal transcribed spacer (ITS2) as a genetic marker. This single tube multiplex PCR allows specific amplification of targeted species and produces rapid and accurate results. The specificity was confirmed by sequencing the ITS2 and partial 16S rRNA genes from ticks collected from Estonia, Latvia, Sweden and Russia. We tested the assay in a large-scale experiment, and a total of 1284 ticks removed from rodents and shrews were successfully identified at species level.

Keywords: I. persulcatus; I. ricinus; I. trianguliceps; ITS2; Species identification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA, Ribosomal Spacer / genetics
Europe, Eastern
Genetic Markers
Ixodes / classification*
Ixodes / genetics*
Multiplex Polymerase Chain Reaction*
Species Specificity
Sweden

Substances

DNA, Ribosomal Spacer
Genetic Markers