Research On CL Immunoassay System Based On Microfluidic Technology

Clenbuterol,used as an animal feed additive,is often used to improve lean meat rates for livestock.People eat meat products with excessive CL,which can appear a series of symptoms such as dizziness,muscle twitch,neuroticism and so on.At present,the technology of detecting clenbuterol hydrochloride content at home and abroad is divided into two major categories.One is the quantitative analysis method of precision instruments,and the other is the method of fast detection.The quantitative analysis method of precision equipment is expensive,and the detection time is long.It has certain requirements for operators,and it is difficult to detect quickly,real-time and mass.Although the dipsticks have the advantages of low cost and short detection time,the detection limit is low and the sensitivity is not high.It is necessary to further confirm the quantitative analysis method of the above precision instrument.This article will start from the perspective of experimental research.Combining microfluidics,ELISA and POCT and Transferring of traditional ELISA kit to microfluidic chips establish a new rapid detection technique for CL residues in large batch field screening.Including:1)Based on ELISA,the analysis and design of the sampling,flow channel,reaction pool and waste water pool of microfluidic chip were analyzed and designed.The material of microfluidic chip is selected from the angle of stability,processing performance and cost performance,and the processing.To theoretical study on the driving method for microscale transport of fluid in micro pipeline and selecte the micro pump valve drive system with higher stability and strong driving ability After comparison of various driving methods.In order to reduce the injection error caused by the peristaltic pump pulsation,a method of accurately controlling the injection volume by time control was designed in this paper,so that the quantitative control performance has been greatly improved.2)Building CL detection hardware platform for microfluidic chip driven by micro pump valve.First of all,the selection of the micro pump valve was fully investigated and the driving structure of the four pump valve was designed.The experimental test on the driving performance of the fluid was carried out;A constant temperature system was set up which to build a controllable constant temperature environment.The portable optical detection device was designed and made to obtain the absorbance analysis with high sensitivity and fast response.3)Experimental studying on biological verification of microfluidic immunoassay and investigating the relationship between CL antigen inclusion,reagent consumption,reagent reaction and color time,washing times,minimum detection limit and other parameters to draw the standard curve of CL.The minimum detection limit was obtained for washing test of micro pump valve.The recovery rate of standard addition was tested,and the reaction time was tested.The stability test and repeated experiment were carried out respectively.The results show that:The optimal coating concentration is 0.25 g/m.The minimum detection limit is 0.025 ng/ml.The detection could be achieved within 24 min,only 1/2 of the traditional time.The detection range of sample concentration was between 0.10 ng/mL and 8.10 ng/ml.The testing equipment is small and easy to carry,and it is 1/4 of the traditional equipment.