Extraction of total nucleic acids from ticks for the detection of bacterial and,

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Extraction of total nucleic acids from ticks for the

detection of bacterial and,viral pathogens.

 

J Med Entomol. 2010 Jan;47(1):89-94.

 

Extraction of total nucleic acids from ticks for the detection of

bacterial and viral pathogens.

 

Crowder CD, Rounds MA, Phillipson CA, Picuri JM, Matthews HE,

Halverson J, Schutzer SE, Ecker DJ, Eshoo MW.

 

Ibis Biosciences, 1896 Rutherford Road, Carlsbad, CA 92008, USA.

 

Ticks harbor numerous bacterial, protozoal, and viral pathogens

that can cause serious infections in humans and domestic animals.

Active surveillance of the tick vector can provide insight into

the frequency and distribution of important pathogens in the

environment. Nucleic-acid based detection of tick-borne

bacterial, protozoan, and viral pathogens requires the extraction

of both DNA and RNA (total nucleic acids) from ticks. Traditional

methods for nucleic acid extraction are limited to extraction of

either DNA or the RNA from a sample.

Here we present a simple bead-beating based protocol for

extraction of DNA and RNA from a single tick and show detection

of Borrelia burgdorferi and Powassan virus from individual,

infected Ixodes scapularis ticks. We determined expected yields

for total nucleic acids by this protocol for a variety of adult

tick species. The method is applicable to a variety of arthropod

vectors, including fleas and mosquitoes, and was partially

automated on a liquid handling robot.

 

Publication Types:

Research Support, N.I.H., Extramural

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

Research Support, U.S. Gov't, P.H.S.

 

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PMID: 20180313 [PubMed - in process]