A novel real-time PCR assay for the speciation of medically important ticks.
The identification of ticks using morphological characters is a well-established practice, however specimens that are small or damaged are often difficult to speciate. A novel, rapid real-time PCR assay, which targets the second internal transcribed spacer (ITS2) region in the nuclear ribosomal DNA gene, was developed for identification of four tick species of utmost medical importance in the United States: Ixodes scapularis, I. pacificus, Dermacentor variabilis, and Amblyomma americanum. Computational analyses of public databases and DNA sequencing studies revealed regions that could be specifically targeted with oligonucleotides optimized for TaqMan chemistry. The oligonucleotide sets designed in this study are specific at both the genus and species levels, and are sensitive at 0.1-1 pg of total tick DNA.
Shone
Scott M
SM
Association of Public Health Laboratories, National Center for Infectious Diseases/Centers for Disease Control and Prevention, Emerging Infectious Diseases Research Fellowship Program, Washington, DC, USA.
Dillon
Harry J
HJ
Hom
Sherman S
SS
Delgado
Nelson
N
D000818:Animals; D001096:Arachnid Vectors / Q000145:classification* / Q000235:genetics; D001483:Base Sequence; D021901:DNA, Intergenic; D021903:DNA, Ribosomal Spacer; D003870:Dermacentor / Q000145:classification; D018884:Ixodes / Q000145:classification; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013987:Ticks / Q000145:classification* / Q000235:genetics
D021901:DNA, Intergenic; D021903:DNA, Ribosomal Spacer