TY - JOUR
T1 - An easy 'one tube' method to estimate viability of Cryptosporidium oocysts using real-time qPCR
AU - Paziewska-Harris, A.
AU - Schoone, G.
AU - Schallig, H. D. F. H.
PY - 2016
Y1 - 2016
N2 - 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
AB - 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
U2 - https://doi.org/10.1007/s00436-016-5044-z
DO - https://doi.org/10.1007/s00436-016-5044-z
M3 - Article
C2 - 27095569
SN - 0932-0113
VL - 115
SP - 2873
EP - 2877
JO - Parasitology Research
JF - Parasitology Research
IS - 7
ER -