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.
Department of Environmental Health and Toxicology, Tokyo Metropolitan Institute of Public Health, Shinjuku, Tokyo, Japan. Akiko_Inomata@member.metro.tokyo.jp
A
A
Inomata
N
N
Kishida
T
T
Momoda
M
M
Akiba
S
S
Izumiyama
K
K
Yagita
T
T
Endo
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