Polymerase chain reaction detection apparatus and polymerase chain reaction detection method

Abstract

The present invention relates to a polymerase chain reaction detection apparatus and a polymerase chain reaction detection method. The polymerase chain reaction detection apparatus is sued for detecting a sample droplet, and comprises a sample conveying mechanism, an excitation light source mechanism, an optical detection mechanism, an impedance detection mechanism and a control mechanism, wherein the impedance detection mechanism comprises an excitation power supply, electrodes and an impedance analyzer, the electrodes are at least two electrodes and respectively comprise a first electrode and a second electrode, the excitation power supply, the first electrode, the second electrode and the impedance analyzer are connected to form a circuit, the first electrode and the second electrode are arranged on the surface of a detection micro-channel and respectively form cross contact with the detection micro-channel, a detection region capable of accommodating at least a sample droplet is formed between the first electrode and the second electrode, and the impedance analyzer is used for collecting the impedance signal produced by the sample droplet passing through the detection region. The polymerase chain reaction detection apparatus of the present invention has high accuracy and high sensitivity.

Description

technical field [0001] The invention relates to the technical field of polymerase chain reaction detection, in particular to a polymerase chain reaction detection device and method. Background technique [0002] Polymerase Chain Reaction (PCR) is a molecular biology technique for amplifying specific nucleic acids in vitro, which has outstanding advantages such as strong specificity, high sensitivity, high efficiency and quickness, and simple operation. Digital PCR is to divide the PCR reaction system containing primers, templates, fluorescent probes and polymerases into many small and independent reaction units. After the PCR amplification reaction, the fluorescence signal of each reaction unit is detected, and the droplets with target DNA fragments will emit strong fluorescence after amplification, that is, positive droplets; the droplets without target DNA fragments will be amplified. Finally, there is only weak background fluorescence, that is, negative droplets; positiv...

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Problems solved by technology

However, the fluorescence of the negative microdroplets is very weak, and there are often missed or wrong detections when detected and analyzed by the fluorescence method.

In addition, when detecting and analyzing microdroplets by the fluorescence method, the identification degree of their characteristic parameters is not high, and there are problems such as fluorescent dye leakage, difficult cleaning of pipelines, and susceptibility to environmental interference, and it is easy to misjudge some abnormal strong fluorescent signal interference as Positive droplet, causing false detection of positive droplet

Therefore, the accuracy and sensitivity of digital PCR based on fluorescence detection are low

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