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Development of a multiple cross displacement amplification combined with nanoparticles-based biosensor assay to

Shijun Li1, Chunting Liu1, Ying Liu1

  • 1Laboratory of Bacterial Infectious Disease of Experimental Center, Guizhou Provincial Center for Disease Control and Prevention, Guiyang, Guizhou, 550004, People's Republic of China.

Infection and Drug Resistance
|August 14, 2019
PubMed
Abstract

Insights

A new rapid assay detects Neisseria meningitidis, the cause of meningococcal disease. This Multiple Cross Displacement Amplification combined with a Lateral Flow Biosensor (MCDA-LFB) offers quick and accurate results.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Neisseria meningitidis is a significant global pathogen causing meningococcal disease.
  • Rapid detection methods are crucial for controlling outbreaks.
  • Existing methods may lack speed or accessibility in certain settings.

Purpose of the Study:

  • To develop and validate a rapid detection assay for Neisseria meningitidis.
  • To utilize Multiple Cross Displacement Amplification (MCDA) and Lateral Flow Biosensor (LFB) technologies.
  • To target the ctrA gene specific to N. meningitidis.

Main Methods:

  • Designed a set of 10 primers targeting 10 distinct regions of the ctrA gene.
  • Developed and optimized the MCDA reaction conditions (64°C for 35 minutes).
  • Integrated MCDA with a Lateral Flow Biosensor (LFB) for direct detection of amplification products.

Main Results:

  • Successfully established an MCDA-LFB assay for N. meningitidis detection.
  • Achieved a rapid turnaround time of under 60 minutes for the entire process.
  • Demonstrated high sensitivity with a detection limit of 10 fg/reaction (approx. 3 CFUs/reaction) and specificity with no cross-reactivity.

Conclusions:

  • The developed MCDA-LFB assay is an effective tool for rapid N. meningitidis detection.
  • This method is particularly valuable for resource-poor settings during meningococcal disease epidemics.
  • The assay provides a fast, sensitive, and specific diagnostic solution.
Keywords:
MCDA-LFBNeisseria meningitidislateral flow biosensorlimit of detectionmultiple cross displacement amplification

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