MicroRNAs
(miRNAs) are novel tumor biomarkers owing to their important
physiological functions in cell communication and the progression
of multiple diseases. Due to the small molecular weight, short sequence
length, and low concentration levels of miRNA, miRNA detection presents
substantial challenges, requiring the advancement of more refined
and sensitive techniques. There is an urgent demand for the development
of a rapid, user-friendly, and sensitive miRNA analysis method. Here,
we developed an enhanced biotin–streptavidin dual-mode phase
imaging surface plasmon resonance (PI-SPR) aptasensor for sensitive
and rapid detection of miRNA. Initially, we evaluated the linear sensing
range for miRNA detection across two distinct sensing modalities and
investigated the physical factors that influence the sensing signal
in the aptamer-miRNA interaction within the PI-SPR aptasensor. Then,
an enhanced biotin–streptavidin amplification strategy was
introduced in the PI-SPR aptasensor, which effectively reduced the
nonspecific adsorption by 20% and improved the limit of detection
by 548 times. Furthermore, we have produced three types of tumor marker
chips, which utilize the rapid sensing mode (less than 2 min) of PI-SPR
aptasensor to achieve simultaneous detection of multiple miRNA markers
in the serum from clinical cancer patients. This work not only developed
a new approach to detect miRNA in different application scenarios
but also provided a new reference for the application of the biotin–streptavidin
amplification system in the detection of other small biomolecules.