Abstract
Five differently preserved groups of adult Rhipicephalus appendiculatus specimens were compared for quality of DNA extracted. Three methods were used to extract DNA from specimens i.e. two simple mosquito validated DNA extraction methods and a tick validated method. Extraction of DNA from tick legs was attempted. The quality of DNA extracted was evaluated by the success of PCR amplification of the ITS2 gene and the mitochondrial COI gene fragment. Fresh specimens (i.e. killed just before extraction) had the highest success of DNA amplification followed by specimens killed in ethanol and subsequently stored in the refrigerator (4 °C). There was no significant difference in amplification success between cryopreserved and 70% ethanol preserved specimens. It was possible to amplify DNA from legs of ticks. Sequenced ITS2 amplicon of template obtained from legs of ticks was as legible as those from whole tick extract. The two mosquito validated DNA extraction methods showed a significantly lower amplification success than the tick validated protocol.
Similar content being viewed by others
References
S.C. Barker (1998) ArticleTitleDistinguishing species and populations of Rhipicephaline ticks with ITS 2 ribosomal RNA J. Parasitol. 84 887–892 Occurrence Handle1:CAS:528:DyaK1cXnt1GgsLs%3D Occurrence Handle9794625
R. Boom C.J.A. Sol M.M.M. Salimans C.L. Jansen P.M.E. Wertheim-van Dillen J. Noordaa ParticleVan Der (1990) ArticleTitleRapid and simple method of purification of nucleic acids J. Clin. Microbiol. 28 495–503 Occurrence Handle1:CAS:528:DyaK3cXhvFCrsbY%3D Occurrence Handle1691208
R. Boom C. Sol M. Beld J. Weel J. Goudsmit P. Wertheim-van Dillen (1999) ArticleTitleImproved silica-guanidiniumthiocyanate DNA isolation procedure based on selective binding of bovine α-casein to silica particles J. Clin. Microbiol. 37 615–619 Occurrence Handle1:CAS:528:DyaK1MXhsleisb0%3D Occurrence Handle9986822
R. Burgemeister R. Gangnus B. Haar K. Schütze U. Sauer (2003) ArticleTitleHigh quality RNA retrieved from samples obtained by using LMPC (Laser Microdisection and Pressure Catapulting) technology Pathol. Res. Pract. 199 431–436 Occurrence Handle10.1078/0344-0338-00442 Occurrence Handle1:CAS:528:DC%2BD3sXot1Sjurw%3D Occurrence Handle12924446
F.H. Collins M.A. Mendez M.O. Rasmussen P.C. Mehaffey N.J. Besansky V. Finnerty (1987) ArticleTitleA ribosomal RNA gene probe differentiates member species of Anopheles gambiae complex Am. J. Trop. Med. Hyg. 37 37–41 Occurrence Handle1:CAS:528:DyaL2sXltFSgs7s%3D Occurrence Handle2886070
A. Crampton I. McKay S.C. Barker (1996) ArticleTitlePhylogeny of ticks (Ixodida) inferred from nuclear ribosomal DNA Int. J. Parasitol. 26 511–517 Occurrence Handle1:CAS:528:DyaK28XkvVCrtL4%3D Occurrence Handle8818731
P.R. Crosbie W.M. Boyce T.C. Rodwell (1998) ArticleTitleDNA sequence variation in Dermacentor hunteriestimated phylogenies of Dermacentor spp. (Acari: Ixodidae) in the New world J. Med. Entomol. 35 277–288 Occurrence Handle1:CAS:528:DyaK1cXjs12hu78%3D Occurrence Handle9615547
R.H. Cruickshank (2002) ArticleTitleMolecular markers for the phylogenetics of mites and ticks Syst. Appl. Acarol. 7 3–14
M.D. Dean J.W.O. Ballad (2001) ArticleTitleFactors affecting DNA quality from museum preserved Drosophila simulans Entomol. Exp. Appl. 98 279–283 Occurrence Handle10.1023/A:1018963131302 Occurrence Handle1:CAS:528:DC%2BD3MXjtFyitrk%3D
N. Dillon A.D. Austin E. Bartowsky (1996) ArticleTitleComparison of preservation techniques for DNA extraction from hymenopterous insects Insect. Mol. Biol. 5 21–24 Occurrence Handle1:CAS:528:DyaK28XhtVylsrs%3D Occurrence Handle8630531
M.J. Domanico R.B. Phillips T.H. Oakley (1997) ArticleTitlePhylogenetic analysis of pacific salmon (genus Oncorhynchus) using nuclear and mitochondrial DNA sequences Can. J. Fish. Aquat. Sci. 54 1865–1872 Occurrence Handle10.1139/cjfas-54-8-1865 Occurrence Handle1:CAS:528:DyaK2sXntlGltbw%3D
A.A. Guglielmone A. Estrada-Peňa A.J. Mangold D.M. Barros-Battesti M.B. Labruna J.R. Martins J.M. Venzal M. Arzua J.E. Keirans (2003) ArticleTitleAmblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch1844 (Acari: Ixodidae): hosts, distribution and 16S rDNA sequences Vet. Parasitol. 113 273–288 Occurrence Handle1:CAS:528:DC%2BD3sXjtlWksrc%3D Occurrence Handle12719142
R. Hernandez A.C. Chen R.B. Davey G.W. Ivie G.G. Wagner J.E. George (1998) ArticleTitleComparison of genomic DNA in various strains of Boophilus microplus (Acari: Ixodidae) J. Med. Entomol. 35 895–900 Occurrence Handle1:CAS:528:DyaK1cXmtlOrurw%3D Occurrence Handle9775626
C.A. Hill J.A. Gutierrez (2003) ArticleTitleA method for extraction and analysis of high quality genomic DNA from ixodid ticks Med. Vet. Entomol. 17 224–227 Occurrence Handle10.1046/j.1365-2915.2003.00425.x Occurrence Handle1:STN:280:DC%2BD3s3ps12hsw%3D%3D Occurrence Handle12823841
M.J. Hubbard K.J. Cann D.J.M. Wright (1995) ArticleTitleValidation and rapid extraction of nucleic acids from alcohol-preserved ticks Exp. Appl. Acarol. 19 473–478 Occurrence Handle1:STN:280:BymC3Mnlt1Y%3D Occurrence Handle8529479
A.J. Mangold M.D. Barques S. Mas-Coma (1998) ArticleTitle18S rRNA gene sequences and phylogenetic relationships of European hard-tick species (Acari: Ixodidae) Parasitol. Res. 84 31–37 Occurrence Handle10.1007/s004360050433 Occurrence Handle1:STN:280:DyaK1c7lt1WhsQ%3D%3D Occurrence Handle9491423
D.K. McLain D.M. Wesson J.H. Olivier SuffixJr. F.H. Collins (1995) ArticleTitleVariation in ribosomal DNA internal transcribed spacers 1 among eastern populations of Ixodes scapularis (Acari: Ixodidae) J. Med. Entomol. 32 353–360 Occurrence Handle1:STN:280:ByqA2c7mtlE%3D Occurrence Handle7616527
A. Murrell N.J.H. Campbell S.C. Barker (2000) ArticleTitlePhylogenetic analyses of the rhipicephaline ticks indicate that the genus Rhipicephalus is paraphyletic Mol. Phylogenet. Evol. 16 1–7 Occurrence Handle1:CAS:528:DC%2BD3cXksVaqt7g%3D Occurrence Handle10877935
A. Murrell N.J.H. Campbell S.C. Barker (2001) ArticleTitleRecurrent gains and losses of large (84–109 bp) repeats in the rDNA internal transcribed spacer 2 (ITS2) of rhipicephaline ticks Insect. Mol. Biol. 10 587–59 Occurrence Handle10.1046/j.0962-1075.2001.00298.x Occurrence Handle1:CAS:528:DC%2BD38Xps1Gl Occurrence Handle11903628
A.J. Phillips C. Simon (1995) ArticleTitleSimpleefficient and non-destructive DNA extraction protocol for arthropods Ann. Entomol. Soc. Am. 88 281–283 Occurrence Handle1:CAS:528:DyaK2MXms12lurY%3D
C. Simon F. Frati A. Beckenbach B. Crespi H. Liu P. Flook (1994) ArticleTitleEvolution, weighting and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers Ann. Entomol. Soc. Am. 87 651–701 Occurrence Handle1:CAS:528:DyaK2MXis1Wiu7g%3D
N. Speybroeck M. Madder P. Bossche ParticleVan Den J. Mtambo N. Berkvens G. Chaka M. Mulumba J. Brandt L. Tirry D. Berkvens (2002) ArticleTitleDistribution and phenology of ixodid ticks in southern Zambia Med. Vet. Entomol. 16 430–441 Occurrence Handle10.1046/j.1365-2915.2002.00397.x Occurrence Handle1:STN:280:DC%2BD3s%2FgsVyktw%3D%3D Occurrence Handle12510896
InstitutionalAuthorNameStata Corp. (2003) Stata/SE 8.0 for Windows statistical software Stata Corporation College Station, Texas
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mtambo, J., Van Bortel, W., Madder, M. et al. Comparison of Preservation Methods of Rhipicephalus appendiculatus (Acari: Ixodidae) for Reliable DNA Amplification by PCR. Exp Appl Acarol 38, 189–199 (2006). https://doi.org/10.1007/s10493-006-0004-4
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10493-006-0004-4