A highly sensitive TaqMan real-time PCR assay for early detection of Schistosoma species

► In order to maximize the detection sensitivity, the sequence of the 18S rRNA gene was chosen as the PCR target gene which is a repeated region with high copy numbers. ► As we using the real-time PCR method, further down-stream analysis is not required, which shortens the time needed to obtain results by conventional electrophoresis on agarose gel. Also TaqMan chemistry was chosen since for high throughput testing, it is not necessary to interrogate melt profiles for every well as would be the case with SYBR Green I. ► S. japonicum egg DNA could be detected by TaqMan real-time PCR one week earlier than the microscopy method could, and the peak percentage of positive animals detected is much higher than that by microscopy. ► Since DNA derived from decaying cercarae corpse or worm eggs cannot last for a long time, the DNA detected by our real-time PCR assay are generated by the parasite in “real-time”. Thus our method is potentially useful for quantification of infection by Asian Schistosoma parasite and for monitoring the burden in infection in patients.