Development Of A Suspension Array Method For Simultaneously Quantitative Detection Of Multi-pathogens

Objective: To establish a platform for rapid, sensitive, specific, high-throughput, and simultaneous detection of multiple biothreat-associated agents, a suspension array-based multiplexed immunoassay was developed for exploring the feasibility of directly simultaneous detection of five pathogens including bacterium, bacterial spore, virus, bacterial toxin, and plant toxin in powder samples.Methods: The 5-plexed immunoassays using sets of 9-plexed coupled fluorescent beads were employed to simultaneously detect five representative biothreat agents, including Y. pestis, B. anthracis spore, staphylococcal enterotoxin B (SEB), ricin, and SARS-CoV, from a single powder sample and the feasibility for detection of them in field samples was demonstrated by both blind and standard laboratory trials. Results: The newly developed suspension array appeared to be specific and sensitive, with the detection sensitivities of about 67.5cfu/mL, 10~3 cfu/mL, 0.203 ng/mL, 29.1ng/mL, 4.86ng/mL for Y. pestis, B. anthracis spore, SEB, ricin, SARS-CoV N protein, respectively, and the dynamic ranges of about 10~2～10~8cfu/mL, 10~3～10~7cfu/mL, 0.02-152.7ng/mL, 3.56-100852.1ng/mL, 0.391-25600ng/mL, respectively, which were higher than those of corresponding conventional ELISA tests.Conclusion: The suspension array could rapidly, sensitively , specifically , simultaneously and quantitatively detect five pathogens from powder samples, which would be useful for early identification, rapid diagnosis, and response for the attack of bioterrorism and outbreak of infectious disease.