Spectrally resolved chemiluminescent probes for sensitive multiplex molecular quantification.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Molecular Probes

Background:

• Fluorescent luminophores are common for analyte detection but distinguishing multiple chemically initiated luminophores is challenging.
• Existing methods for multiplexing chemically initiated luminophores are limited, hindering their broader application.

Purpose of the Study:

• To develop a new class of hybrid luminophore probes for sensitive, multiplexed detection.
• To enable rapid, rational design of spectrally resolvable, chemically initiated probes.

Main Methods:

• Developed hybrid luminophore probes with covalently conjugated fluorophores and an acridinium ester chemiluminophore.
• Utilized energy transfer (ET) for light emission at distinct wavelengths and intensities.
• Performed time-resolved spectrographic and luminescence characterizations.

Main Results:

• Demonstrated that ET is not dependent on spectral overlap, suggesting a non-Förster resonance mechanism.
• Showcased the probes' ability for sensitive, multiplex quantification of nucleic acid sequences.
• Achieved detection of environmentally relevant microbes without prior enzymatic amplification.

Conclusions:

• The novel hybrid luminophore probes facilitate multiplexed detection of analytes.
• This approach offers a sensitive and efficient method for quantifying microbial nucleic acids.
• The findings advance the development of chemically initiated probes for various applications.