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Detection of Cryptosporidium parvum oocysts using a microfluidic device equipped with the SUS micromesh and FITC‐labeled antibody
Author(s) -
Taguchi Tomoyuki,
Arakaki Atsushi,
Takeyama Haruko,
Haraguchi Satoshi,
Yoshino Masato,
Kaneko Masao,
Ishimori Yoshio,
Matsunaga Tadashi
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21104
Subject(s) - cryptosporidium parvum , cryptosporidium , microfluidics , antibody , microbiology and biotechnology , biology , chemistry , virology , chromatography , nanotechnology , materials science , feces , immunology
Development of a microfluidic device equipped with micromesh for detection of Cryptosporidium parvum oocyst was reported. A micromesh consisting of 10 × 10 cavities was microfabricated on the stainless steel plate by laser ablation. Each cavity size, approximately 2.7 µm in diameter, was adopted to capture a single C. parvum oocyst. Under negative pressure operation, suspensions containing microbeads or C. parvum oocysts flowed into the microchannel. Due to strong non‐specific adsorption of microbeads onto the PDMS microchannel surface during sample injection, the surface was treated with air plasma, followed by treatment with 1% sodium dodecyl sulfate (SDS) solution. This process reduced the non‐specific adsorption of microbeads on the microchannel to 10% or less in comparison to a non‐treated microchannel. This microfluidic device equipped with the SUS micromesh was further applied for the capture of C. parvum oocysts. Trapped C. parvum oocysts were visualized by staining with FITC‐labeled anti‐ C. parvum oocyst antibody on a micromesh and counted under fluoroscopic observation. The result obtained by our method was consistent with that obtained by direct immunofluorescence assay coupled with immunomagnetic separation (DFA‐IMS) method, indicating that the SUS micromesh is useful for counting of C. parvum oocysts. The newly designed microfluidic device exploits a geometry that allowed for the entrapment of oocysts on the micromesh while providing the rapid introduction of a series of reagents and washes through the microfluidic structure. Our data indicate that this microfluidic device is useful for high‐throughput counting of C. parvum oocysts from tap water sample. Biotechnol. Bioeng. 2007;96: 272–280. © 2006 Wiley Periodicals, Inc.