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Updated: Jun 14, 2026
Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres
Published on: October 23, 2011
[Development of a universal primers PCR-coupled liquid bead array to detect biothreat bacteria]
Hai-yan Wen1, Jing Wang, Heng-chuan Liu
1Institute of Health Quarantine, China Academy of Inspection and Quarantine, Beijing, China.
Objective:
To develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.
Methods:
16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed.
Results:
After PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples.
Conclusion:
The suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.
