Closed-tube DNA extraction using a thermostable proteinase is highly sensitive, capable of single parasite detection.
Current DNA extraction methods for parasites are labour-intensive and usually involve several steps, increasing the potential for cross-contamination. We describe here a closed-tube DNA extraction procedure based upon the use of a thermostable proteinase that enabled sensitive amplification of target loci from parasites from diverse lineages including Apicomplexa, Sarcomastgophora and Nematoda. Moreover, this procedure is not subject to cross-contamination and is readily adaptable to automation.
Department of Chemistry and Biomolecular Sciences, Environmental Biotechnology CRC and Biotechnology Research Institute, Macquarie University Sydney, Sydney, NSW, 2109, Australia. bferrari@cbms.mq.edu.au
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D000818:Animals; D002417:Cattle; D016054:DNA, Protozoan / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004795:Enzyme Stability; D005243:Feces / Q000469:parasitology; D005434:Flow Cytometry; D005821:Genetic Techniques; D005849:Gerbillinae / Q000469:parasitology; D006801:Humans; D018189:Immunomagnetic Separation; D010271:Parasites / Q000235:genetics* / Q000302:isolation & purification*; D010447:Peptide Hydrolases / Q000378:metabolism*; D013696:Temperature
D016054:DNA, Protozoan; D010447:Peptide Hydrolases