Microfluidic immunoassay for rapid detection of cotinine in saliva
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Microfluidic immunoassay for rapid detection of cotinine in saliva Kaiping Cheng & Wang Zhao & Sixiu Liu & Guodong Sui
# Springer Science+Business Media New York 2013
Abstract A microfluidic immunoassay is successfully developed for rapid analysis of cotinine saliva samples, which is a metabolite of nicotine and is widely used as a biomarker to evaluate the smoking status and exposure to tobacco smoke. The core microfluidic chip is fabricated by polydimethylsiloxane (PDMS) with standard soft lithography. Each chip is capable of eight parallel analyses of cotinine samples. The analyses can be completed within 40 min with 12 μl sample consumption. The linear detection range is 1~250 ng/ml and the minimum detectable concentration is 1 ng/ml respectively. The correlation coefficient of the calibration curve established from standard samples is 0.9989. The immunoassay was also validated by real saliva samples, and the results showed good reproducibility and accuracy. All the results were confirmed with traditional ELISA measurements. The result from microfluidic chip device and ELISA kits showed good correspondence, and the correlation coefficients are higher than 0.99. Compared with traditional technique, this microfluidic immunoassay is more economic, rapid, simple and sensitive, perfect for on-site cotinine measurements as well as for the evaluation of the exposure to tobacco smoking. Moreover, this immunoassay has potential to be applied in the analysis of other biomarkers in human saliva samples. Keywords Cotinine . Microfluidic device . Saliva . Passive smoking . Active smoking . ELISA
K. Cheng : W. Zhao : S. Liu : G. Sui (*) Department of Environmental Science and Engineering, Institute of Biomedical Science, Fudan University, Shanghai 200433, People’s Republic of China e-mail: [email protected] G. Sui Shanghai Tobacco Group, NO. 717, Changyang Road, Shanghai 200082, People’s Republic of China
1 Introduction It is well known that smoking is harmful to human health and environment, more than 4000 chemical components have been identified in the smoke, and about 60 of them are suspicious carcinogens (Adams and Hamilton 2008; Adhikari, et al. 2009; Thielen, et al. 2008). These chemicals may cause cancer (Bennett, et al. 1999; Chen, et al. 2003), apogeny (Pasinska, et al. 2009; Stekhun et al. 1979), cardiovascular (Ezzati, et al. 2005; Thun, et al. 1999) and cerebrovascular diseases (Ezzati, et al. 2005; Lederle, et al. 2003; Mazzone, et al. 2010; Silvestrini, et al. 1996), and many other diseases (Braithwaite, et al. 2012; Austin, et al. 2002; Bisanovic, et al. 2011; Ueshima, et al. 2004; Heitzer and Meinertz 2005). During smoking, two kinds of smoke with different composition are produced: mainstream smoke which is inhaled by the active smoker and sidestream smoke released into the environment through puffs from the lit end of the cigarette (Thielen, et al. 2008). Compared with mainstream smoke, sidestream smoke was produced in larger quantities and contained 2 times more nicotine and 12 times more ammonia per c
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