Fusion genes, playing
a causal role in human tumorigenesis and
developments, are deemed as gold standard molecular biomarkers in
cancer diagnosis, therapy, and prognosis. A rapid, robust, and sensitive
method of detection of fusion genes for point-of-care (POC) diagnosis
is urgently needed. Here, taking the advantages of the superior specificity
of the ligation reaction and the highly amplified efficiency of isothermal
exponential amplification with a pH indicator, we developed a colorimetric
method for visual detection of fusion genes with high sensitivity
and specificity by the naked eye. More importantly, we first found
that fusion genes can be accurately quantified in a wide dynamic range
(2 zmol to 2 fmol) by an open-source app with a smartphone-assisted
RGB (red, green, and blue value) reading mode. The proposed method
for Visual detection of Fusion genes by Ligation-triggered Isothermal
Exponential Amplification is termed Vis-Fusion LIEXA. We have demonstrated
that the Vis-Fusion LIEXA is a practical and reliable method for accurate
quantitative detection of the fusion gene in a complex biological
sample at zmol level in 40 min only with a smartphone, thereby providing
a user-friendly and point-of-care testing (POCT) tool for molecular
diagnostics.
A method based on mRNA-templated ligation of splice-junction anchored DNA probes followed by PCR amplification of the ligated product has been developed for multiplexed detection of mRNA splice variants with...
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