The development of a qPCR assay to detect tick (Ixodida) DNA and its implementation for the study of tick-borne pathogen transmission.
Polymerase chain reaction (PCR) analysis is regularly used to detect pathogens within arthropod vectors, but has also been applied to investigate vector DNA. This study details a novel highly sensitive quantitative PCR (qPCR) which detects and quantifies DNA from Ixodes ricinus, the European vector of Anaplasma phagocytophilum. By pairing this with a qPCR to detect A. phagocytophilum, valid comparisons of pathogen load can be made between different sized tick-tissue samples. These qPCRs were validated in I. ricinus that were fed A. phagocytophilum-infected blood using an artificial membrane feeder. Pathogens were detected in the tick haemolymph within 36h, indicating that successful infection had taken place. This study illustrates the application of vector-targeted qPCRs to confirm and validate pathogen load in samples as part of investigations of vector-pathogen interactions.
School of Clinical Veterinary Sciences, University of Bristol, Langford House, Langford, North Somerset, UK. m.robinson@bristol.ac.uk
M T
MT
Robinson
E R
ER
Morgan
D
D
Woods
S E
SE
Shaw
D041081:Anaplasma phagocytophilum / Q000235:genetics / Q000302:isolation & purification; D000818:Animals; D001096:Arachnid Vectors / Q000235:genetics* / Q000382:microbiology; D004247:DNA / Q000302:isolation & purification*; D005260:Female; D018884:Ixodes / Q000235:genetics* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D017282:Tick-Borne Diseases / Q000635:transmission*
D004247:DNA