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; Miren Iturriza-Go'Mara; 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, Miren Iturriza-Go'Mara, R. M. Chalmers^*

^*Corresponding author for this work

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

34 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 - Apr 2012

Bibliographical note

Funding Information:
We thank Jonathan Goss at the UK Cryptosporidium Reference Unit for performing the Method 1 DNA extractions. Hannah Fairclough was supported by a Students into Work grant from the Society for Applied Microbiology, who also part-funded this work.

Keywords

Cryptosporidium
DNA extraction
Real time PCR

Access to Document

10.1016/j.mimet.2012.02.006

Cite this

@article{0fe7cc3046f6402f8a25dc21b9c56e2b,

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

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.",

keywords = "Cryptosporidium, DNA extraction, Real time PCR",

author = "K. Elwin and G. Robinson and Hadfield, {S. J.} and Fairclough, {H. V.} and Miren Iturriza-Go'Mara and Chalmers, {R. M.}",

note = "Funding Information: We thank Jonathan Goss at the UK Cryptosporidium Reference Unit for performing the Method 1 DNA extractions. Hannah Fairclough was supported by a Students into Work grant from the Society for Applied Microbiology, who also part-funded this work.",

year = "2012",

month = apr,

doi = "10.1016/j.mimet.2012.02.006",

language = "English",

volume = "89",

pages = "38--40",

journal = "Journal of Microbiological Methods",

issn = "0167-7012",

publisher = "Elsevier B.V.",

number = "1",

}

TY - JOUR

T1 - A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay

AU - Elwin, K.

AU - Robinson, G.

AU - Hadfield, S. J.

AU - Fairclough, H. V.

AU - Iturriza-Go'Mara, Miren

AU - Chalmers, R. M.

N1 - Funding Information: We thank Jonathan Goss at the UK Cryptosporidium Reference Unit for performing the Method 1 DNA extractions. Hannah Fairclough was supported by a Students into Work grant from the Society for Applied Microbiology, who also part-funded this work.

PY - 2012/4

Y1 - 2012/4

N2 - 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.

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

UR - http://www.scopus.com/inward/record.url?scp=84857773468&partnerID=8YFLogxK

U2 - 10.1016/j.mimet.2012.02.006

DO - 10.1016/j.mimet.2012.02.006

M3 - Article

C2 - 22366300

AN - SCOPUS:84857773468

SN - 0167-7012

VL - 89

SP - 38

EP - 40

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

IS - 1

ER -

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

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Cite this