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Updated: Jan 14, 2026
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
A Real-Time PCR-Based Method for the Sequence Determination of Cryptosporidium hominis and Cryptosporidium parvum
Guy Robinson1, Kristin Elwin2, Rachel M Chalmers2
1Cryptosporidium Reference Unit, Public Health Wales Microbiology and Health Protection, Singleton Hospital, Swansea, Wales, UK. Guy.Robinson@wales.nhs.uk.
Abstract:
The gp60 gene has been widely used to subtype Cryptosporidium for many years. Predominantly, this has involved nested polymerase chain reaction (PCR) amplification, gel electrophoresis, and Sanger sequencing of the amplicons. However, laboratories may no longer have gel electrophoresis capability, and during outbreak investigations a more timely approach may be required. Here, we describe a real-time PCR assay combined with high-resolution melt curve analysis that streamlines the PCR detection of gp60 amplicons of Cryptosporidium parvum and Cryptosporidium hominis for sequencing during public health investigations.
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