Digital PCR microarray image analysis method

Abstract

The invention provides a digital PCR (Polymerase Chain Reaction) microarray image analysis method, which comprises the following steps of: inputting a first channel fluorescence image, and preprocessing the first channel fluorescence image; carrying out image segmentation on the preprocessed fluorescence image; identifying effective units of the segmented image, and extracting centroid coordinates of the effective units; positioning and extracting effective units in the second and third channel images according to the coordinates, and counting fluorescence signal intensity of the effective units in the second and third channel images according to the effective units; according to the signal intensity, drawing a scatter diagram and a histogram of the fluorescence intensity of the reaction units in the second channel image and the third channel image, counting the proportion of the positive units, combining Poisson distribution, and calculating the concentration of the target nucleic acid molecules in the original reaction liquid. The mask is established by the first channel image so as to indirectly analyze the second channel image and the third channel image, so that the multi-channel digital PCR microarray image under non-uniform illumination can be effectively analyzed, the accuracy of image analysis is improved, and absolute quantitative analysis of target nucleic acid molecules is realized.

Description

technical field [0001] The invention relates to the field of image processing, in particular to a digital PCR microarray image analysis method. Background technique [0002] Digital PCR is a nucleic acid detection technology with high sensitivity, high specificity and absolute quantification. Using the idea of ​​"divide and conquer", the PCR reaction system is subdivided into tens of thousands of micro-reaction systems, so that all reaction systems can be carried out independently After the PCR cycle amplification reaction, after the reaction is completed, the proportion of positive units is counted, and the concentration of the target nucleic acid molecule in the original reaction solution can be calculated by using the Poisson distribution. In recent years, with the continuous development of chip manufacturing technology, chips containing tens of thousands of micro-reaction units have been manufactured through micro-processing technology. The chip in the QuantastudioTM3D ...

AI Technical Summary

Problems solved by technology

However, for images with uneven illumination and blurred images, the accuracy of image segmentation is low.

[0003] Therefore, in view of the problems of inaccurate segmentation, low recognition rate and weak robustness in the current digital PCR image analysis technology, it is of great significance to develop a high-accuracy digital PCR microarray image analysis method.

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