Droplet Digital PCR (ddPCR)

Digital polymerase chain reaction (dPCR) is a biotechnological refinement of conventional polymerase chain reaction methods that can be used to directly quantify and clonally amplify nucleic acids strands including DNA, cDNA, or RNA. The key difference between dPCR and traditional PCR lies in the method of measuring nucleic acids amounts, with the former being a more precise method than PCR. Like PCR, in dPCR, a single reaction is also performed within a sample, but the sample is divided into numerous partitions, with the reaction being performed in each partition separately. With this division, nucleic acid quantities may be collected and measured more accurately and sensitively. The technique is regularly used for clonal amplification of materials for next-generation sequencing and has shown beneficial for analysing changes in gene sequences, including copy number variants and point mutations. Droplet Digital PCR (ddPCR) is a technique for dPCR in which a 20-microliter sample reaction is split into 20,000 nanoliter-sized oil droplets using a water-oil emulsion technique, thermocycled in a microfluidic cartridge, and the fluorescence amplitude of each droplet in each sample well is read using a droplet scanner.

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Applications of Droplet Digital PCR

Clinical Diagnostics

· Qualitative/quantitative detection of infectious pathogenic microorganisms (especially low-copy microorganisms), detection of mutation in pathogen resistance gene, pathogen typing and identification.
· Tumor/cancer-related gene variation (mutation, fusion, methylation, CNV, etc.) for early cancer screening, companion diagnosis, targeted drug therapy, treatment guidance, treatment efficacy monitoring and evaluation.
· Non-invasive prenatal screening, detection of neonatal genetic diseases such as congenital hypothyroidism and phenylketonuria.

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Environmental Surveillance

· Detection of complex and low abundance microorganisms in water, soil, gas, and other components.

· Detection of drug-resistant infectious pathogenic microorganisms in wards, laboratories, hospitals, and other environmental surfaces.

Food Safety

· Detection of pathogenic/non-pathogenic microorganisms in foods such as dairy products.

· Food sampling and inspection, such as meat adulteration, and genetically modified food ingredients.

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Biopharmaceuticals/ Gene Therapy

· Efficiency assessment of gene editing.

· Biological safety assessment of vaccines and antibodies for residues of host-derived substances.
· Drug efficacy evaluation.

Forensic Science

· Species identification and quantification of samples.

· Genetic marker analysis for

identification.
· Analysis of trace evidence and inference of time of death.

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Agriculture

· Identification of genetically modified species.
· Stress resistance gene analysis.
· Research on genetic differences between species.
· Plant breeding.
· Detection/identification of source of diseases, such as identification of parasites and pathogenic microorganisms.

Preparation of Standard Materials

· Quantitation and quality control of reference materials.