Fluorescent Microspheres Immunochromatographic Assays For Rapidly Detection Of Prealbumin And Bone Alkaline Phosphatase

The diseases may break out when malnutrition existent in humans. Through the literature review and market research, we learned that prealbumin(PA) and bone alkaline phosphatase(BAP) as the specific biochemical markers of detection protein-energy nutritional status and calcium nutritional status, respectively, can monitor the people’s nutritional status stably and accurately. The research aimed to develop the fluorescence immunochromatographic assay for rapid and quantitative detection of the PA and BAP in serum, respectively. The established method would used in clinical application and provide the basic data for monitoring the nutritional status of people.The immunochromatographic assays were proposed for detecting PA in serum by using sandwich principle. Based on the PA concentration in the specimens was proportional to the fluorescence intensity ratio of the test line and the control line(FIT/FIC), quantitative analysis has been achieved. In this study, anti-PA monoclonal antibody(anti-PA m Ab) and polyclonal antibody(anti-PA p Ab) were paired based on double-antibody sandwich. Probe including fluorescent microspheres and antibody was prepared by EDC one step-mediated method. The experimental parameters in labeling process and the test strip preparation process were optimized, and the optimal conditions was selected to detect the PA in the specimen. Results showed that the limit of detection of PA reached 1.0 ng/m L within 20 min with a good linear range of 8.0 ng/m L to 110.0 ng/m L(R2 = 0.998). The test strip showed no cross reaction with hemoglobin and albumin. The variation coefficients of intra- and inter-assay were 3.3 % to 4.3 % and 4.1 % to 9.9 %, respectively. A significantly good agreement was observed between the test strip and immunoturbidimetric assay(R2 =0.94). The developed novel assay in this study is a sensitive, specific, time-saving for detection of human PA in serum.The immunochromatographic test strip assay based on fluorescent microspheres probe for detection of BAP in serum was also established in the study. Theexperimental parameters were optimized according to the result of the test strip. The test strip was evaluated on the basis of the optimum conditions. The results showed that the test strip had good accuracy and stability while the detection time was 15 min.The sensitivity was 0.1 ng/m L, and the assay exhibited good linear range from 0.1ng/m L to 250 ng/m L with reliable correlation of 0.997. The correlation coefficient between the test strip and chemiluminescence immunoassay was 0.771.Nutritional status monitoring in clinical is usually time-consuming, and also need a variety of instruments. The developed immunochromatographic assay in this study has the advantages of sensitive, specific, time-saving, and quantitative.Therefore, the assay has high application values.