Quantitative mRNA expression measurement at home.

Autor: Pandey S; Shanvi, San Diego, CA, USA.; Department of Pediatrics, University of California San Diego, 9500 Gilman Drive, MC 0703, Leichtag Building 132, La Jolla, CA, 92093-0703, USA., McCoy SS; Department of Pediatrics, University of California San Diego, 9500 Gilman Drive, MC 0703, Leichtag Building 132, La Jolla, CA, 92093-0703, USA.; Department of Computer Science and Engineering, Jacob's School of Engineering, University of California San Diego, La Jolla, USA., Stobdan T; Department of Pediatrics, University of California San Diego, 9500 Gilman Drive, MC 0703, Leichtag Building 132, La Jolla, CA, 92093-0703, USA., Sahoo D; Shanvi, San Diego, CA, USA. dsahoo@ucsd.edu.; Department of Pediatrics, University of California San Diego, 9500 Gilman Drive, MC 0703, Leichtag Building 132, La Jolla, CA, 92093-0703, USA. dsahoo@ucsd.edu.; Department of Computer Science and Engineering, Jacob's School of Engineering, University of California San Diego, La Jolla, USA. dsahoo@ucsd.edu.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Jan 10; Vol. 14 (1), pp. 1013. Date of Electronic Publication: 2024 Jan 10.
DOI: 10.1038/s41598-023-49651-8
Abstrakt: mRNA measurement is dominated by RT-PCR, which requires expensive laboratory equipment and personnel with advanced degrees. Loop-mediated isothermal amplification (LAMP) is a versatile technique for detecting target DNA and RNA. The sensitivity of LAMP in early reports has been below that of the standard RT-PCR tests. Here, we report the use of a fluorescence-based RT-LAMP protocol to measure CDX2 expression patterns, which match extremely well to the standards of sophisticated RT-PCR techniques (r = 0.99, p < 0.001). The assay works on diverse sample types such as cDNA, mRNA, and direct tissue sample testing in 25 min compared to more than 3 h for RT-PCR. We have developed a new protocol for designing RT-LAMP primers that reduce false positives due to self-amplification and improve quantification. A simple device with a 3D-printed box enables the measurement of mRNA expression at home, outdoors, and point-of-care setting.
(© 2024. The Author(s).)
Databáze: MEDLINE
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