Screening Ticks from Different Counties and Comparing Them
1407 Words6 Pages
Ticks will be collected from areas in the five counties in the state where Lyme disease and other tick-borne infections are prevalent. Trapping sites will be set in known tick habitats in the five counties to help in trapping and collecting small animals, including the target pests. Sherman live-traps baited with oatmeal and peanut-butter will be set at the sites to ensnare the ticks. All captured animals will be regularly and carefully examined to allow the researchers to extract as many ticks as possible. The trapping of animals and extraction of ticks from the captured animals will take about four weeks.
All ticks collected will be placed in vials containing 75% ethanol and sent to parasitology laboratories. The screening process will be done through MasTag polymerase chain reaction (PCR), a multiplication method, which is known for its fast and economic screening of tick-borne pathogens. PCR is also known for efficient and accurate screening of ticks for polymicrobial contagion. Before beginning the PCR procedure, the ticks will be removed from the vials, washed with saline containing phosphate buffer, and then homogenized with Tri-reagent LS.
The next step will involve nucleic acid extraction, where total RNA will be suspended in about 30ml of H2O. The generation of cDNA will then follow where 20ml of the solution and 15ml of total RNA will be reacted using Superscript 11 Reverse Transcriptase as a catalyst. The MassTag PCR procedure will then be done using the tick panel, with primers added to the reactants to help detect the Borrelia species responsible for relapsing fever and the Powassan virus. All the MassTag PCR assays that will be done in the experiment will utilize 3ml of cDNA. The final PCR products wi...
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