An On-Site Serology Monitoring System Consisting of a Multiplex Microfluidic Chip Fabricated Using the Electrospray Deposition Method for Laboratory Mice

  • AOKI Hiroyoshi
    Ultra High Precision Fabrication Team, Advanced Technology Support Division, Advanced Science Institute, RIKEN Ultra High Precision Fabrication Team, Advanced Technology Support Division, Advanced Science Institute, RIKEN
  • KANEKO Ai
    Fuence Co., Ltd. Fuence Co., Ltd.
  • KAJITA Ayako
    Experimental Animal Division, RIKEN BioResource Center Experimental Animal Division, RIKEN BioResource Center
  • YAMAGATA Yutaka
    Ultra High Precision Fabrication Team, Advanced Technology Support Division, Advanced Science Institute, RIKEN Ultra High Precision Fabrication Team, Advanced Technology Support Division, Advanced Science Institute, RIKEN
  • IKE Fumio
    Experimental Animal Division, RIKEN BioResource Center Experimental Animal Division, RIKEN BioResource Center
  • KASE Hiroshi
    Fuence Co., Ltd. Fuence Co., Ltd.

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Abstract

Mice are important laboratory animals used in biological and medical studies. Pathogen infections in laboratory mice are detected using the enzyme-linked immunosorbent assay (ELISA), which requires a large quantity of serum. A low serum requirement is important for more frequent and detailed analyses for infections. A new serology monitoring system with a 16-microchannel microfluidic chip was developed to identify anti-pathogen antibodies in 0.2 µL of mouse serum; the system indicates infections caused by 6 important murine pathogens: Salmonella enterica serotype Typhimurium, Sendai virus, mouse hepatitis virus, Mycoplasma pulmonis, lymphocytic choriomeningitis virus, and ectromelia virus. To avoid non-specific adsorption of mouse sera to the microchannel, a polyvinyl alcohol-based blocking method was developed. To equalize the sensitivities of different antigens, the quantities of pathogen antigens deposited were optimized through precision spotting by using the electrospray deposition method. A comparative study with the conventional ELISA method was carried out, and the developed microfluidic chip system was able to simultaneously and specifically detect antibodies against the above mentioned 6 pathogens with good linearity (R2=0.955–0.966) in 12 min with a lower serum quantity (1/13), reaction time (1/16), and antigen quantities (1/19–1/2,800) than the conventional ELISA. Validation using a serum panel (n=501) also showed good agreement with previous results.

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