Closed-tube DNA extraction using a thermostable proteinase is highly sensitive,capable of single parasite detection |
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| Authors: | B. C. Ferrari M. L. Power P. L. Bergquist |
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| Affiliation: | (1) Department of Chemistry and Biomolecular Sciences, Environmental Biotechnology CRC and Biotechnology Research Institute, Macquarie University Sydney, Sydney, NSW, 2109, Australia;(2) Department of Biological Sciences, Macquarie University Sydney, Sydney, NSW, 2109, Australia;(3) Department of Molecular Medicine & Pathology, University of Auckland Medical School, Auckland, New Zealand |
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| Abstract: | 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. |
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| Keywords: | DNA extraction Cryptosporidium Giardia Haemonchus Nematoda Proteinase Protozoa |
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