Digital pcr: a technology review

Halimeh Rezaei ,1,* Sheyda khalilian,2

1. Division of Genetics, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran, IR Iran
2. Division of Genetics, Department of Biology, Faculty of Science, University of Isfahan, Isfahan, Iran, IR Iran

Abstract

Introduction

Any permanent alteration of the nucleotide sequences in the genome is considered as a gene mutation. gene mutations are closely related to the occurrence and progression of many disorders; therefore the mutation identification is a significant part of genetic screening of the disorders. several methods have been developed to detect mutations such as real-time quantitative pcr. in recent years, digital pcr (dpcr), as the third generation of pcr, has been developed for the absolute quantification of target nucleic acids. the advent of dpcr, elevates the detection of gene mutations, especially in cancer-associated genes. this technology has been utilized in the detection of tumor markers in cell-free dna (cfdna) samples from patients with different types of cancer and in this way has a significant role in basic research and clinical applications. in this review we try to summarize the dpcr applications in the detection of gene mutations related to human diseases.

Methods

The search was done in electronic databases such as pubmed, elsevier, and scopus to introduce the most recent studies.

Results

In the dpcr, the sample is divided in to different independent partitions and amplified target sequences are detected by fluorescence. the presence of fluorescence allows determining the concentration of the target in the sample. dpcr is highly used in detecting pathogen mutations, analysis of gene expression, assessment of genome editing effects, screening cancer biomarkers,… and it is necessary to state that the gene mutation analysis is an important aspect for all these fields.

Conclusion

Although dpcr is a novel technology that performs a key role in clinical applications, it has some limitations such as the large sample volume and highly allele-specific probes. the higher accuracy and sensitivity of dpcr in compare with qpcr, indicates a great advantage in the detection of rare mutations. altogether, the advent technology of dpcr can help to promote the development of personalized cancer treatment, gene-modifying techniques, mitochondrial disease management and hopefully in near future, it can be applied in disease-specific tailored treatments and adopted as a routine clinical assay.

Keywords

Digital pcr, dpcr, gene mutation, real-time pcr, genome editing