Detection of Cryptosporidium parvum in buffer and in complex matrix using PEMC sensors at 5 oocysts mL(-1).
We show for the first time that a piezoelectric-excited millimeter-sized cantilever (PEMC) sensor can detect five Cryptosporidium parvum oocysts in 25% milk in PBS background in a flow format (1 mL min(-1)). To improve sensitivity, a secondary antibody (murine IgM) was used to confirm the attachment of oocysts to the sensor. PEMC sensor is a resonant-mode cantilever sensor whose resonant frequency decreases when target analyte binds to its surface. The sensor was functionalized with Protein G, and then immobilized with goat polyclonal IgG anti-C. parvum for oocysts detection. In the dynamic range of 50-10,000 oocysts mL(-1) the sensor response is characterized by a semi-log relationship between resonant frequency response and C. parvum oocysts concentration. In 25% milk background, binding kinetics was slower and total sensor response was lower (approximately 45%) than in water-like buffer.
Department of Chemical and Biological Engineering, Drexel University, 3141 Chestnut St., Philadelphia, PA 19104, USA.
Sen
S
Xu
Raj
R
Mutharasan
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D015374:Biosensing Techniques / Q000295:instrumentation; D002021:Buffers; D016785:Cryptosporidium parvum / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M; D007700:Kinetics; D057230:Limit of Detection; D051379:Mice; D033661:Oocysts; D014873:Water Pollutants / Q000032:analysis
D001426:Bacterial Proteins; D002021:Buffers; C034284:IgG Fc-binding protein, Streptococcus; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D014873:Water Pollutants