In the past few years, paper-based inkjet printing biosensor and bioanalytical tools have been extensively used for the rapid detection of biomolecule interactions. Many research activities increasingly concentrate on microfluidic devices fabricated with glass and polymer surfaces, which have attracted great attention because of their potential miniature form and automation. Specifically, studies have validated these paper-based microfluidic biosensors, i.e., surface-modified miniaturized microfluidic devices, as a novel analytical tool for sequential analytical measurements. It includes chemiluminescent methods1, surface-Raman spectroscopy2, electrochemical3, and FRET-based fluorescent detection methods4–6. On the other hand, the design and fabrication of these microfluidic devices would be complex, highly expensive, and time- consuming. Hence, a growing need exists for a cost-effective and similar method for the detection of biomolecules. Nitrocellulose (NC) membranes7,8, filter paper9, parchment paper9, chromatographic paper8, or glass fiber paper-like6 substrate have been utilized as the paper-based material, because they possess high protein/enzyme binding capability that makes it available for bio-molecular immobilization. These porous membranes or paper operates based on capillary action to transport and react with liquid samples. So far, paper-based inkjet printing microfluidic sensors have been employed for sandwich ELISAs7, α-amylase detector for disease diagnosis10, detection of acetylcholinesterase (AChE) inhibitors11, and micro-colorimetric biochemical (glucose/glucose oxidase, DNA/hydrogen peroxidase and biotin/streptavidin) detection method12. Similarly, automated paper-based inkjet printing sandwich ELISA was fabricated on a piece of nitrocellulose membrane to analyze human chronic gonadotropin (hCG). However, this method also includes multiple steps, and crucial printing patterns are required to obtain quantitative outcomes7. Different paper-based microfluidic devices have reported FRET-based fluorescent assay for the direct detection of protein, nucleic acid, and upconversion phosphors (UCPs) suitable for molecular diagnosis4,6. Recently, portable paper-based sensor bis (dithiocarbamato) copper (II) complex functionalized carbon nanodots (CDs) for the detection of mercuric ion (Hg2+) were developed by printing CuDTC2-CD solution on cellulose acetate paper using a commercial inkjet printer5. Although various inkjet-printing paper-based ELISA platforms with colorimetric detection for drug screening, and molecular diagnosis and enzyme inhibitory analysis have been successfully developed in the recent years, a paper-based inkjet-printing technique is still not widely applied for FRET detection. Paper-based assays are commonly utilized for detecting biologically small molecules and macromolecules because of their effective accessibility and fewer false-positive results13–16. A variety of applications based on paper and inkjet printing-based diagnosis have been reported for molecular diagnosis17–23, RNA detection and analysis for Ebola virus diagnosis13, C-reactive protein (CRP) monitoring24, multiplexed point-of-care diagnostic devices to detection of nucleic acids, malaria and dengue14–16,25,26.