用于聚合酶链反应的低成本三维编程微型泵

Low Cost Three-Dimensional Programmed Mini-Pump Used in PCR.

作者信息

Lin Chengxiong, Wang Yaocheng, Huang Zhengyu, Guo Yu, Wu Wenming

机构信息

National Engineering Research Center for Healthcare Devices, Guangdong Provincial Key Laboratory of Medical Electronic Instruments and Polymer Material Products, Institute of Biological and Medical Engineering, Guangdong Academy of Sciences, Guangzhou 510316, China.

School of Mechanical and Electrical Engineering, Guangdong University of Technology, Guangzhou 510006, China.

出版信息

Micromachines (Basel). 2022 May 14;13(5):772. doi: 10.3390/mi13050772.

DOI:10.3390/mi13050772

PMID:35630239

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9143699/

Abstract

Programmed mini-pumps play a significant role in various fields, such as chemistry, biology, and medicine, to transport a measured volume of liquid, especially in the current detection of COVID-19 with PCR. In view of the cost of the current automatic pipetting pump being higher, which is difficult to use in a regular lab, this paper designed and assembled a three-dimensional programmed mini-pump with the common parts and components, such as PLC controller, motor, microinjector, etc. With the weighting calibration before and after pipetting operation, the error of the pipette in 10 μL (0.2%), 2 μL (1.8%), and 1 μL (5.6%) can be obtained. Besides, the contrast test between three-dimensional programmed mini-pump and manual pipette was conducted with the ORF1ab and pGEM-3Zf (+) genes in qPCR. The results proved that the custom-made three-dimensional programmed mini-pump has a stronger reproducibility compared with manual pipette (ORF1ab: 24.06 ± 0.33 vs. 23.50 ± 0.58, = 0.1014; pGEM-3Zf (+): 11.83.06 ± 0.24 vs. 11.50 ± 0.34, = 0.8779). These results can lay the foundation for the functional, fast, and low-cost programmed mini-pump in PCR or other applications for trace measurements.

摘要

程控微型泵在化学、生物学和医学等各个领域发挥着重要作用，用于输送定量体积的液体，特别是在当前通过聚合酶链式反应（PCR）检测新冠病毒（COVID-19）的过程中。鉴于目前自动移液泵成本较高，难以在常规实验室中使用，本文采用可编程逻辑控制器（PLC）、电机、微量注射器等常见零部件设计并组装了一种三维程控微型泵。通过移液操作前后的重量校准，可得出该移液器在10微升（误差0.2%）、2微升（误差1.8%）和1微升（误差5.6%）时的误差。此外，在实时荧光定量PCR（qPCR）中，使用开放阅读框1ab（ORF1ab）基因和pGEM-3Zf（+）基因对三维程控微型泵和手动移液器进行了对比测试。结果证明，与手动移液器相比，定制的三维程控微型泵具有更强的重复性（ORF1ab：24.06±0.33对23.50±0.58，P = 0.1014；pGEM-3Zf（+）：11.83±0.24对11.50±0.34，P = 0.8779）。这些结果可为PCR或其他痕量测量应用中功能化、快速且低成本的程控微型泵奠定基础。