Digital polymerase chain reaction optical detection device and method

Abstract

The invention relates to a digital polymerase chain reaction optical detection device and method. The device for detecting sample microdroplets comprises a sample transmission mechanism, an excitation light source mechanism, an optical detection mechanism, an input display mechanism and a control mechanism; the sample transmission mechanism is provided with a transmission micro channel for transmitting the sample microdroplets and a detection micro channel, and the detection micro channel is arranged at one end of the transmission micro channel and is only used for single row of the sample microdroplets to pass through; the excitation light source mechanism is provided with an excitation light source for exciting the sample microdroplets to generate an optical signal; the optical detection mechanism is provided with an optical fiber spectrometer for collecting and converting the optical signal of the sample microdroplets; the input display mechanism is used for setting or displaying the passing speed of the sample microdroplets in the sample transmission mechanism, the luminous intensity of the excitation light source and the spectral data of the sample microdroplets; the control mechanism is used for controlling the passing speed of the sample microdroplets in the sample transmission mechanism and the luminous intensity of the excitation light source. The device has the advantages of simple structure and high detection accuracy.

Description

technical field [0001] The invention relates to the technical field of polymerase chain reaction detection, in particular to a digital polymerase chain reaction optical detection device and method. Background technique [0002] Polymerase Chain Reaction (PCR) is a molecular biology technique for amplifying specific nucleic acids in vitro. Among them, digital PCR (Digital Polymerase Chain Reaction, dPCR) is the third generation PCR technology, digital PCR is to divide the PCR reaction system containing primers, templates, fluorescent probes, polymerase and other components into many tiny, independent reaction units . Each reaction unit contains a nucleic acid template as much as possible. After PCR amplification reaction, the fluorescence signal of each reaction unit is detected and statistically analyzed to realize the absolute quantitative analysis of nucleic acid. [0003] According to the different ways of forming the reaction unit, digital PCR is mainly divided into th...

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Problems solved by technology

[0004] The droplet digital PCR optical detection device in the traditional technology is mainly composed of several mechanisms such as an excitation light source, a sample microchannel, and a weak light detector. The detected optical signals include fluorescent signals and scattered light signals, etc. Sensor detection, the optical signal collection optical path is more complicated, which increases the instability of the mechanism

In addition, a variety of fluorescent probes are used in droplet digital PCR, and multiplex PCR requires multiple excitation light sources and multi-window fluorescence signal detection. The optical signals between different fluorescent probes and between fluorescence and excitation light sources will vary Mutual interference, the true intensity of the target fluorescence is difficult to measure, and the detection accuracy is low

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