Validation of droplet digital Polymerase Chain Reaction for the detection and absolute quantification of Taenia solium eggs in spiked soil samples
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Date
2019
Journal Title
Journal ISSN
Volume Title
Publisher
Elsevier
Abstract
To enable the detection of taeniid eggs in environmental samples, a sensitive technology is required. In this
study, we validated the effectiveness of a digital droplet Polymerase Chain Reaction (ddPCR) assay for detection,
identification and absolute quantification of taeniid DNA from artificially contaminated soils with varying
numbers of taeniid eggs using a set of universal primers, JB3 & JB4.5. The results showed that the number of
cox1 copies detected increased gradually for both species with the number of taeniid eggs added to the different
soil types. The highest cox1 DNA copies recovery for Taenia solium and T. lynciscapreoli was from the sand soil
with lowest recovery being observed in clay soils. Therefore, ddPCR is a promising technology for screening of
taeniid eggs from soil samples collected in the environment irrespective of the soil type and the number of eggs.
The potential of the ddPCR protocol to detect taeniid egg DNA in spiked soil samples has great practical application
for taeniid egg screening in soils from endemic areas. However, when universal primers are used in
screening environmental samples, the identity of ddPCR positive samples must be confirmed by sequencing. In
addition, more validation studies using species-specific primers and field soil samples is recommended.
Description
Journal article
Keywords
Soil, ddPCR, Taeniid eggs, Taenia solium, Taenia lynciscapreoli, Detection