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Clinical Chemistry 0: clinchem.2006.073569v1, 2006; 10.1373/clinchem.2006.073569
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Received on May 18, 2006
Accepted on September 29, 2006

Automation and Analytical Techniques

Rapid and Simultaneous Quantification of 4 Urinary Proteins by Piezoelectric Quartz Crystal Microbalance Immunosensor Array

Yang Luo 1, Ming Chen 1, Qianjun Wen 2, Meng Zhao 1, Bo Zhang 1, Xiaoyan Li 3, Feng Wang 1, Qing Huang 1, Chunyan Yao 1, Tianlun Jiang 1, Guoru Cai 4, Weiling Fu 1*

1 Department of Laboratory Medicine, Southwest Hospital, Third Military Medical University, Chong Qing, People's Republic of China
2 Laboratory Center of Urology, Southwest Hospital, Third Military Medical University, Chong Qing, People's Republic of China
3 Endocrinology Department, Southwest Hospital, Third Military Medical University, Chong Qing, People's Republic of China
4 The 26th Research Institute, Chinese Electronic Scientific and Technical Group Company, Chong Qing, People's Republic of China

* To whom correspondence should be addressed. E-mail: weilingfu{at}yahoo.com.

Background: Urinary proteins are predictive and prognostic markers for diabetes nephropathy. Conventional methods for the quantification of urinary proteins, however, are time-consuming, and most require radioactive labeling. We designed a label-free piezoelectric quartz crystal microbalance (QCM) immunosensor array to simultaneously quantify 4 urinary proteins.

Methods: We constructed a 2 x 5 model piezoelectric immunosensor array fabricated with disposable quartz crystals for quantification of microalbumin, {alpha}1-microglobulin, {beta}2-microglobulin, and IgG in urine. We made calibration curves after immobilization of antibodies at an optimal concentration and then evaluated the performance characteristics of the immunosensor with a series of tests. In addition, we measured 124 urine samples with both QCM immunosensor array and immunonephelometry to assess the correlation between the 2 methods.

Results: With the QCM immunosensor array, we were able to quantify 4 urinary proteins within 15 min. This method had an analytical interval of 0.01-60 mg/L. The intraassay and interassay imprecisions (CVs) were <10%, and the relative recovery rates were 90.3%-109.1%. Nonspecificity of the immunosensor was insignificant (frequency shifts <20 Hz). ROC analyses indicated sensitivities were ≥95.8% and, specificities were ≥76.3%. Bland-Altman difference plots showed the immunosensor array to be highly comparable to immunonephelometry.

Conclusions: The QCM system we designed has the advantages of being rapid, label free, and highly sensitive and thus can be a useful supplement to commercial assay methods in clinical chemistry.




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