An easy 'one tube' method to estimate viability of Cryptosporidium oocysts using real-time qPCR

Paziewska-Harris, A. ORCID: https://orcid.org/0000-0001-9677-4753, Schoone, G. and Schallig, H. D. F. H. 2016. An easy 'one tube' method to estimate viability of Cryptosporidium oocysts using real-time qPCR. Parasitology Research 115 (7) , pp. 2873-2877. 10.1007/s00436-016-5044-z

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Abstract

Viability estimation of the highly resistant oocysts of Cryptosporidium remains a key issue for the monitoring and control of this pathogen. We present here a simple ‘one tube’ quantitative PCR (qPCR) protocol for viability estimation using a DNA extraction protocol which preferentially solubilizes excysted sporozoites rather than oocysts. Parasite DNA released from excysted sporozoites was quantified by real-time qPCR using a ribosomal DNA marker. The qPCR signal was directly proportional to the number of oocysts excysted, and a power-law relationship was noted between oocyst age and the proportion excysting. Unexcysted oocysts released negligible amounts of DNA making the method suitable for estimating viability of as few as 10 oocysts.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Publisher:	Springer Verlag
ISSN:	0932-0113
Date of First Compliant Deposit:	12 April 2019
Date of Acceptance:	7 April 2016
Last Modified:	28 Nov 2024 17:30
URI:	https://orca.cardiff.ac.uk/id/eprint/121740

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