玻璃结构PCR芯片快速精密温控及实验验证
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摘要
PCR芯片作为即时检测技术的一种核心生化分析器件,在疾病快速现场检测、便携式分析中有着广泛应用。温度快速精密控制对提高PCR芯片的扩增效率和准确性极为重要。相较于聚合物材料,玻璃材料具备优良的生物兼容性、导热性和密封性。设计并制作了一种全玻璃结构的PCR芯片,以PT100温度传感器和恒流源组成温度检测电路,将温度值线性转换成电压值。采用STM32控制器作为核心处理器,完成电压值的数字化采集。结合积分分离PID算法,对加热电极和风扇施加驱动电流和控制电压,实现对PCR芯片的快速精密温度控制。实验结果表明,该PCR芯片可获得6℃/s的升温速率以及3℃/s的降温速率,控温精度为0.4℃,优于当前市场上大多数PCR仪的性能。生物扩增实验验证了PCR芯片DNA扩增的有效性。
As a core device for biochemical analysis in point-of-care testing(POCT)technology,PCR chip is widely used in rapid on-site disease detection and portable analysis.Rapid and precise temperature control is extremely important to improve the amplification efficiency and accuracy of PCR chip.Compared to polymer materials,glass materials have excellent biocompatibility,thermal conductivity and sealing properties.A whole glass structure PCR chip was designed and fabricated.The temperature detection circuit is composed of PT100 temperature sensor and constant current source,which can linearly convert temperature into voltage.Using STM32 controler as the core processor to acquire the digital voltage,the system applies integral separation PID algorithm to apply driving current and control voltage to heating electrode and fan,thus fast and precise temperature control of PCR chip can be realized.Eexperimental verification shows that the PCR chip can obtain a heating and cooling rate of 6 and3℃/s,respectively,and the temperature control accuracy is 0.4℃.Bio-amplification experiment verifies the effectiveness of PCR chip for DNA amplification.
As a core device for biochemical analysis in point-of-care testing(POCT)technology,PCR chip is widely used in rapid on-site disease detection and portable analysis.Rapid and precise temperature control is extremely important to improve the amplification efficiency and accuracy of PCR chip.Compared to polymer materials,glass materials have excellent biocompatibility,thermal conductivity and sealing properties.A whole glass structure PCR chip was designed and fabricated.The temperature detection circuit is composed of PT100 temperature sensor and constant current source,which can linearly convert temperature into voltage.Using STM32 controler as the core processor to acquire the digital voltage,the system applies integral separation PID algorithm to apply driving current and control voltage to heating electrode and fan,thus fast and precise temperature control of PCR chip can be realized.Eexperimental verification shows that the PCR chip can obtain a heating and cooling rate of 6 and3℃/s,respectively,and the temperature control accuracy is 0.4℃.Bio-amplification experiment verifies the effectiveness of PCR chip for DNA amplification.
引文
[1]曾照芳,洪秀华.临床检验仪器[M].北京:人民卫生出版社,2007.Zeng Zhaofang,Hong Xiuhua.Clinical Laboratory Instruments[M].Beijing:People's Medical Publishing House,2007.
[2]Wang J,Ahmad H,Ma C,et al.A self-powered,one-step chip for rapid,quantitative and multiplexed detection of proteins from pinpricks of whole blood[J].Lab.on a Chip,2010,10:3157-3162.
[3]Mullis K,Faloona F,Scharf S,et al.Specific enzymatic amplification of DNA in vitro:the polymerase chain reaction[J].Cold Spring Harb Symp.Quant.Biol.,1986,51:263-273.
[4]Ouyang Y,Duarte G R M,Poe B L,et al.A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control[J].Anal.Chim.Acta.,2015,901:59-67.
[5]Shaw K J,Docker P T,Yelland J V,et al.Rapid PCRamplification using a microfluidic device with integrated microwave heating and air impingement cooling[J].Lab.on a Chip,2010,10:1725-1728.
[6]Chen J J,Hsieh I H.Using an IR lamp to perform DNAamplifications on an oscillatory thermocycler[J].App.Ther.Eng.,2016,106:1-12.
[7]Fermer C,Nilsson P,Larhed M.Microwave-assisted highspeed PCR[J].Eur.Jour.Phar.Sci.,2003,18:129-132.
[8]Maltezos G,Johnston M,Scherera A.Thermal management in microfluidics using micro-Peltier junctions[J].Appl.Phys.Lett.,2005,87(15):105-108.
[9]殷学锋,沈宏,方肇伦.制造玻璃微流控芯片的简易加工技术[J].分析化学,2003,31(1):116-119.Yin Xuefeng,Shen Hong,Fang Zhaolun.A simplified microfabrication technology for production of glass microfluidic chips[J].Chin.J.Anal.Chem.,2003,31(1):116-119.
[10]王磊,李桂香,王元麟.基于PT100热电阻的温度检测系统设计[J].仪器仪表学报,2014,12(4):33-35.Wang Lei,Li Guixiang,Wang Yuanlin.Temperature measurement system based on PT100[J].China Instrumentation,2014,12(4):33-35.
[11]罗毅,施云波,渠立亮,等.谐波分析及交流比较的高空温度探测方法研究[J].仪器仪表学报,2014,35(4):721-729.Luo Yi,Shi Yunbo,Qu Liliang,et al.Research on high altitude temperature detection method based on harmonic analysis and AC comparison[J].China Instrumentation,2014,35(4):721-729.
[12]刘金琨.先进PID控制MATLAB仿真[M].北京:电子工业出版社,2004.Liu Jinkun.Advanced PID Control MATLAB Simulation[M].Beijing:Publishing House of Electronics Industry,2004.
[2]Wang J,Ahmad H,Ma C,et al.A self-powered,one-step chip for rapid,quantitative and multiplexed detection of proteins from pinpricks of whole blood[J].Lab.on a Chip,2010,10:3157-3162.
[3]Mullis K,Faloona F,Scharf S,et al.Specific enzymatic amplification of DNA in vitro:the polymerase chain reaction[J].Cold Spring Harb Symp.Quant.Biol.,1986,51:263-273.
[4]Ouyang Y,Duarte G R M,Poe B L,et al.A disposable laser print-cut-laminate polyester microchip for multiplexed PCR via infra-red-mediated thermal control[J].Anal.Chim.Acta.,2015,901:59-67.
[5]Shaw K J,Docker P T,Yelland J V,et al.Rapid PCRamplification using a microfluidic device with integrated microwave heating and air impingement cooling[J].Lab.on a Chip,2010,10:1725-1728.
[6]Chen J J,Hsieh I H.Using an IR lamp to perform DNAamplifications on an oscillatory thermocycler[J].App.Ther.Eng.,2016,106:1-12.
[7]Fermer C,Nilsson P,Larhed M.Microwave-assisted highspeed PCR[J].Eur.Jour.Phar.Sci.,2003,18:129-132.
[8]Maltezos G,Johnston M,Scherera A.Thermal management in microfluidics using micro-Peltier junctions[J].Appl.Phys.Lett.,2005,87(15):105-108.
[9]殷学锋,沈宏,方肇伦.制造玻璃微流控芯片的简易加工技术[J].分析化学,2003,31(1):116-119.Yin Xuefeng,Shen Hong,Fang Zhaolun.A simplified microfabrication technology for production of glass microfluidic chips[J].Chin.J.Anal.Chem.,2003,31(1):116-119.
[10]王磊,李桂香,王元麟.基于PT100热电阻的温度检测系统设计[J].仪器仪表学报,2014,12(4):33-35.Wang Lei,Li Guixiang,Wang Yuanlin.Temperature measurement system based on PT100[J].China Instrumentation,2014,12(4):33-35.
[11]罗毅,施云波,渠立亮,等.谐波分析及交流比较的高空温度探测方法研究[J].仪器仪表学报,2014,35(4):721-729.Luo Yi,Shi Yunbo,Qu Liliang,et al.Research on high altitude temperature detection method based on harmonic analysis and AC comparison[J].China Instrumentation,2014,35(4):721-729.
[12]刘金琨.先进PID控制MATLAB仿真[M].北京:电子工业出版社,2004.Liu Jinkun.Advanced PID Control MATLAB Simulation[M].Beijing:Publishing House of Electronics Industry,2004.