Development and evaluation of a reverse transcription-loop-mediated isothermal amplification assay for rapid and high-sensitive detection of Cryptosporidium in water samples.
We describe a novel assay for simple, rapid and high-sensitive detection of Cryptosporidium oocysts in water samples using a reverse transcription-loop-mediated isothermal amplification (RT-LAMP). The assay is based on the detection of 18S rRNA specific for Cryptosporidium oocysts. The detection limit of the developed RT-LAMP assay was as low as 6 x 10(-3) oocysts/test tube, which theoretically enables us to detect a Cryptosporidium oocyst and perform duplicated tests even if water samples contain only one oocyst. The developed RT-LAMP assay could more sensitively detect Cryptosporidium oocysts in real water samples than the conventional assay based on microscopic observation.
Inomata
A
A
Department of Environmental Health and Toxicology, Tokyo Metropolitan Institute of Public Health, Shinjuku, Tokyo, Japan. Akiko_Inomata@member.metro.tokyo.jp
Kishida
N
N
Momoda
T
T
Akiba
M
M
Izumiyama
S
S
Yagita
K
K
Endo
T
T
D000818:Animals; D003458:Cryptosporidium / Q000235:genetics* / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D033661:Oocysts / Q000166:cytology; D048348:Reverse Transcription / Q000235:genetics*; D013696:Temperature; D014867:Water / Q000469:parasitology*
D014867:Water