Workflow diagram of the nano-dLAMP detection system
Digital polymerase chain reaction (digital PCR, dPCR), a high-sensitivity, absolute quantitative and high-tolerance nucleic acid detection technology, plays an important role in a variety of application fields, such as rare mutations detection, copy number variation, liquid biopsy, single cell analysis, genetically modified detection, viral load testing, and microbiological quantitative analysis. In this study, the researchers fabricated microarray chips with four physical partitions for the simultaneous detection of calreticulin type 1, calreticulin type 2, janus kinase 2 V617F mutations and an internal reference gene.
They then used PCR additives and nanoparticles to make the traditional loop mediated isothermal amplification (LAMP) suitable for nanoliter-scale amplification. Results suggested that nanoparticles could improve the amplification performance of nanoliter LAMP. Quantitative detection of the main myeloproliferative neoplasm molecular markers could be performed simultaneously in one four-partition microarray chip within one hour. The agreement between the developed platform and the commercial Quantstudio 3D was high at 99%.
This accurate, rapid, multiplex, and inexpensive nano-dLAMP platform could be a promising tool for clinical diagnosis in the future, according to the researchers. In the digital PCR research and development, chip-based dPCR (cdPCR) method with high isolation stability, good temperature uniformity and fast detection speed was developed. A series of progress were obtained on the detection method, chip, reagent and instrument of cdPCR.
Guojun Cao et al, Establishment of scalable nanoliter digital LAMP technology for the quantitative detection of multiple myeloproliferative neoplasm molecular markers, Sensors and Actuators B: Chemical (2021). DOI: 10.1016/j.snb.2021.130493