A novel biosensing strategy for identification of three important bacteria causing meningitis

Area of Science:

• Molecular Biology
• Biotechnology
• Medical Diagnostics

Background:

• Bacterial meningitis is a severe infection with high mortality, necessitating rapid and accurate diagnostic tools.
• Current diagnostic methods may lack the speed or multiplexing capability required for timely intervention.
• The causative agents Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae are significant public health concerns.

Purpose of the Study:

• To design and evaluate a homemade multiplex PCR assay for simultaneous detection of three major bacterial meningitis pathogens.
• To develop and validate a novel fluorescence biosensor on chip (FBC) for detecting these same pathogens.
• To compare the diagnostic performance, including sensitivity and specificity, of the developed PCR and FBC methods.

Main Methods:

• Fabrication of the FBC involved depositing lead nanoparticles on a quartz slide via thermal evaporation, followed by probe immobilization.
• A multiplex PCR was designed to target specific DNA sequences of Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae.
• Limit of detection (LOD) studies were conducted for both methods using serial dilutions of bacterial strains, with results assessed using a Cytation 5 Cell Imaging Multimode Reader.

Main Results:

• The homemade multiplex PCR demonstrated effective simultaneous detection of all three target bacteria.
• The FBC achieved sensitive detection with varying limits of detection for each pathogen, showing promise for rapid diagnostics.
• Both the multiplex PCR and FBC exhibited appropriate sensitivity and specificity for the detection of Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae.

Conclusions:

• The developed homemade multiplex PCR and novel FBC are viable tools for the rapid and simultaneous diagnosis of bacterial meningitis.
• These diagnostic approaches offer improved sensitivity and specificity compared to some existing methods.
• The FBC, in particular, presents a promising platform for point-of-care meningitis diagnostics.