Amplicon Competition Enables End-Point Quantitation of Nucleic Acids Following Isothermal Amplification.
Yu Sherry JiangApollo StacyMarvin WhiteleyAndrew D EllingtonSanchita BhadraPublished in: Chembiochem : a European journal of chemical biology (2017)
It is inherently difficult to quantitate nucleic acid analytes with most isothermal amplification assays. We developed loop-mediated isothermal amplification (LAMP) reactions in which competition between defined numbers of "false" and "true" amplicons leads to order of magnitude quantitation by a single endpoint determination. These thresholded LAMP reactions were successfully used to directly and quantitatively estimate the numbers of nucleic acids in complex biospecimens, including directly from cells and in sewage, with the values obtained closely correlating with qPCR quantitations. Thresholded LAMP reactions are amenable to endpoint readout by cell phone, unlike other methods that require continuous monitoring, and should therefore prove extremely useful in developing one-pot reactions for point-of-care diagnostics without needing sophisticated material or informatics infrastructure.
Keyphrases
- nucleic acid
- loop mediated isothermal amplification
- sensitive detection
- ms ms
- mass spectrometry
- solid phase extraction
- induced apoptosis
- liquid chromatography tandem mass spectrometry
- liquid chromatography
- cell cycle arrest
- tandem mass spectrometry
- high performance liquid chromatography
- cell therapy
- stem cells
- bone marrow
- deep learning
- big data
- signaling pathway
- cell proliferation
- artificial intelligence
- quantum dots
- antibiotic resistance genes
- mesenchymal stem cells