Development and evaluation of a reverse transcription-loop-mediated isothermal amplification assay for rapid and high-sensitive detection of Cryptosporidium in water samples

摘要

We describe a novel assay for simple, rapid and high-sensitive detection of Cryptosporidiumnoocysts in water samples using a reverse transcription-loop-mediated isothermal amplificationn(RT-LAMP). The assay is based on the detection of 18S rRNA specific for Cryptosporidium oocysts.nThe detection limit of the developed RT-LAMP assay was as low as 6 ￡ 1023 oocysts/test tube,nwhich theoretically enables us to detect a Cryptosporidium oocyst and perform duplicated testsneven if water samples contain only one oocyst. The developed RT-LAMP assay could morensensitively detect Cryptosporidium oocysts in real water samples than the conventional assaynbased on microscopic observation.