Development of immunomagnetic droplet-based digital immuno-PCR for the quantification of prostate specific antigen

Zhou, Wenhan

doi:10.1039/c8ay00921j

Cited by 12 publications

(8 citation statements)

References 22 publications

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“…63 Another approach based on nucleic acid amplification is to perform immuno-PCR on single immunocomplexes associated with beads: Zhou proposed this concept with the beads isolated in droplets. 64 While they did not report immunoassays, Ramsey and co-workers demonstrated digital detection of beads of PCR products from single primer labels on beads sealed in arrays of microwells. 65 Liu and co-workers have shown proof-of-concept of another single molecule approach that localizes fluorescence on individual beads, and allows on-and off-beads to be identified using flow cytometry.…”

Section: Single Molecule Labelsmentioning

confidence: 99%

Digital detection of proteins

Duffy

2023

Lab Chip

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Section: Single Molecule Labelsmentioning

confidence: 99%

Digital detection of proteins

Duffy

2023

Lab Chip

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“…ddPCR can be integrated into iPCR for enabling protein detection with high sensitivity (Ref. 113). EV proteins are profiled using iddPCR amplification method, allowing multiplexed protein profiling of a single EV.…”

Section: Protocols For Isolation and Detection Of Extracellular Vesiclesmentioning

confidence: 99%

“…Immune-droplet digital PCR (iddPCR) is the latest utilisation of PCR technology. ddPCR can be integrated into iPCR for enabling protein detection with high sensitivity (Ref 113)…”

mentioning

confidence: 99%

Extracellular vesicle isolation, purification and evaluation in cancer diagnosis

Mortezaee

Majidpoor

Fathi

2022

Expert Rev. Mol. Med.

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Strategies for non-invasive biomarker discovery in early detection of cancer are an urgent need. Extracellular vesicles (EVs) have generated increasing attention from the scientific community and are under intensive investigations due to their unique biological profiles and their non-invasive nature. EVs are membrane-enclosed vesicles with variable sizes and function. Such vesicles are actively secreted from multiple cell types and are considered as key vehicles for inter-cellular communications and signalling. The stability and potential to easily cross biological barriers enable EVs for exerting durable effects on target cells. These along with easy access to such vesicles, the consistent secretion from tumour during all stages of tumorigenesis and their content providing a reservoir of molecules as well as mirroring the identity of the cell of origin are virtues that have made EVs appealing to be assessed in liquid biopsy approaches and for using as a promising resource of biomarkers in cancer diagnosis and therapy and monitoring targeted cancer therapy. Early detection of EVs will guide time-scheduled personalised therapy. Surveying reliable and sensitive methods for rapid isolation of EVs from biofluids, the purity of isolated vesicles and their molecular profiling and marker specification for clinical translation in patients with cancer are issues in the area and the hot topics of many recent studies. Here, the focus is over methods for EV isolation and stratification for digging more information about liquid biopsy-based diagnosis. Extending knowledge regarding EV-based strategies is a key to validate independent patient follow-up for cancer diagnosis at early stages and inspecting the efficacy of therapeutics.

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“…Similar to the digital readout of single enzyme molecule labels,adigital immuno-PCR can be implemented if the PCR reaction is enclosed in microdroplets.T he droplet-based digital immuno-PCR (ddIPCR) used magnetic beads as as olid support, DNAa sam arker and PCR for signal amplification, for example,f or the detection of PSA. [58] The ddIPCR was performed in three steps as shown in Figure 4. First, the PCR reaction mixture was emulsified to generate tens of thousands of water-in-oil droplets per microchip.…”

Section: Dna Labels For Pcr Amplificationmentioning

confidence: 99%

Advances in Optical Single‐Molecule Detection: En Route to Supersensitive Bioaffinity Assays

et al. 2020

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The ability to detect low concentrations of analytes and in particular low‐abundance biomarkers is of fundamental importance, e.g., for early‐stage disease diagnosis. The prospect of reaching the ultimate limit of detection has driven the development of single‐molecule bioaffinity assays. While many review articles have highlighted the potentials of single‐molecule technologies for analytical and diagnostic applications, these technologies are not as widespread in real‐world applications as one should expect. This Review provides a theoretical background on single‐molecule—or better digital—assays to critically assess their potential compared to traditional analog assays. Selected examples from the literature include bioaffinity assays for the detection of biomolecules such as proteins, nucleic acids, and viruses. The structure of the Review highlights the versatility of optical single‐molecule labeling techniques, including enzymatic amplification, molecular labels, and innovative nanomaterials.

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Development of immunomagnetic droplet-based digital immuno-PCR for the quantification of prostate specific antigen

Abstract: Immuno-PCR (IPCR) is an immunoassay that employs DNA as the signal generator and utilizes both the versatility of antigen–antibody interactions and the exponential amplification power of PCR.

Cited by 12 publications

References 22 publications

Digital detection of proteins

Digital detection of proteins

Extracellular vesicle isolation, purification and evaluation in cancer diagnosis

Advances in Optical Single‐Molecule Detection: En Route to Supersensitive Bioaffinity Assays

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