A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay

K. Elwin; G. Robinson; S. J. Hadfield; H. V. Fairclough; M. Iturriza-Gomara; R. M. Chalmers

doi:10.1016/j.mimet.2012.02.006

A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay

K. Elwin, G. Robinson, S. J. Hadfield, H. V. Fairclough, M. Iturriza-Gomara, R. M. Chalmers

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

33 Citations (Scopus)

Abstract

Direct extraction of Cryptosporidium DNA from 46 stools by bead-beating, guanidine thiocyanate and silica purification provided slightly lower PCR positivity (93.5% vs. 100%) and higher threshold cycle values (mean 34.93 vs. 28.03; P = 0.00) than spin-column extraction from boiled, semi-purified oocyst suspensions. However, direct extraction is cheaper, and amenable to automation.

Original language	English
Pages (from-to)	38-40
Number of pages	3
Journal	Journal of Microbiological Methods
Volume	89
Issue number	1
DOIs	https://doi.org/10.1016/j.mimet.2012.02.006
Publication status	Published - 01 Apr 2012

Keywords

Cryptosporidium
DNA extraction
Real time PCR

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abstract = "Direct extraction of Cryptosporidium DNA from 46 stools by bead-beating, guanidine thiocyanate and silica purification provided slightly lower PCR positivity (93.5% vs. 100%) and higher threshold cycle values (mean 34.93 vs. 28.03; P = 0.00) than spin-column extraction from boiled, semi-purified oocyst suspensions. However, direct extraction is cheaper, and amenable to automation.",

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AU - Iturriza-Gomara, M.

AU - Chalmers, R. M.

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AB - Direct extraction of Cryptosporidium DNA from 46 stools by bead-beating, guanidine thiocyanate and silica purification provided slightly lower PCR positivity (93.5% vs. 100%) and higher threshold cycle values (mean 34.93 vs. 28.03; P = 0.00) than spin-column extraction from boiled, semi-purified oocyst suspensions. However, direct extraction is cheaper, and amenable to automation.

KW - Cryptosporidium

KW - DNA extraction

KW - Real time PCR

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A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay

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Keywords

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