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Very low abundance protein molecular detection,especially for early diagnosis of cancer markers detection is extremely difficult,with the development of single-cell omics in the recent years,there is no a better approach for precise quantification of protein molecules within a single cell.We intend to develop an novel digital proximity ligation assay chip for single protein molecule detection,which integrates the proximity ligation assay for protein detection with the self-priming compartmentalization digital PCR chip to develop an absolute quantitative method for single protein molecule counting-digital proximity ligation assay.This method can provide a new tool for single molecule proteins detection of low abundance proteins and absolute quantification of single cell protein molecule detection.With the advantages of high detection sensitivity,strong specificity,low sample consumption,simple operation,testing equipment common and so on,the digital PLA method not only can detect trace amounts of tumor protein markers in the serum with tiny nonspecific detection,and the number of protein molecules in a single cell by single molecule counting.Compared to the existing single molecule enzyme-linked immunoassays detection technology,it is more simple,less sample consumption,better sensitivity and specificity.For the early diagnosis of cancer,the occurrence and development of tumor,drug screening,single cell analysis and identification of cancer stem cells,digital PLA will have important application value and practical value.