Study On Temperature Calibration System Of Nucleic Acid Isothermal Amplification Instruments

With the increasing demand for rapid nucleic acid detection,the application of nucleic acid isothermal amplification instrument is more than before.Nucleic acid amplification is very sensitive to temperature,so that the isothermal amplification instrument on the temperature performance is strict.Temperature calibration is used to ensure the accuracy and consistency of the sample holes temperature,but the relevant domestic standards are not perfect,so it is necessary to establish a complete set of temperature calibration system.Firstly,this paper introduces the temperature field structure of the nucleic acid isothermal amplification instruments,analyzes the influence of each module on the reagent temperature,and obtains the law of temperature distribution in three-dimensional space at constant temperature of sample seat.Isothermal amplification instruments heating and cooling fast,high temperature control accuracy,so the requirements of the measurement tool must have multi-channel,fast response,high accuracy,good stability characteristics.This paper introduces a multi-channel temperature calibration system,composed of NTC(Negative Temperature Coeffcient,NTC)thermistor temperature sensors,a multi-channel data collector and computer softwares,which can display multiplex temperature in real time,analyze temperature performance,calibrate temperature sensors,and provides necessary assistance for temperature measurement and calibration of isothermal amplification instruments.The temperature of each sample hole of isothermal amplification instruments is not exactly the same,and the selection of acquisition points directly affects the measurement result of temperature performance.According to the principle of heat transfer,this paper puts forward the measurement method of different acquisition points for different temperature parameters,and determines the optimum acquisition point through the measured analysis.Finally,the calibration application of a 8-hole isothermal amplification instrument shows that the system can meet the practical application requirements,the temperature measurement range is 0~120 ℃,the resolution is 0.01 ℃,the acquisition time interval is 1s,and the expanded uncertainty is 0.142 ℃(k=2).The blank about temperature calibration of nucleic acid isothermal amplification instruments in China is filled.