Rapid and simultaneous quantification of 4 urinary proteins by piezoelectric quartz crystal microbalance immunosensor array.
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, alpha1-microglobulin, beta2-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 > or =95.8% and, specificities were > or =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.
Luo
Yang
Y
Department of Laboratory Medicine, Southwest Hospital, The Third Military Medical University, Chongqing, People's Republic of China.
Chen
Ming
M
Wen
Qianjun
Q
Zhao
Meng
M
Zhang
Bo
B
Li
Xiaoyan
X
Wang
Feng
F
Huang
Qing
Q
Yao
Chunyan
C
Jiang
Tianlun
T
Cai
Guoru
G
Fu
Weiling
W
D000510:Alpha-Globulins / Q000652:urine*; D015374:Biosensing Techniques; D002138:Calibration; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D007074:Immunoglobulin G / Q000652:urine*; D046228:Microarray Analysis; D011507:Proteinuria / Q000652:urine*; D011791:Quartz; D012016:Reference Values; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000032:analysis*
D000510:Alpha-Globulins; D007074:Immunoglobulin G; D012709:Serum Albumin; C025500:alpha(2)-microglobulin; C016001:alpha-1-microglobulin; D011791:Quartz