Method and kit for detection of anti-zika virus antibodies

A method, comprising:

• contacting a first portion of a sample from a subject with a first suspension comprising a first plurality of microspheres, wherein the sample comprises blood, serum, plasma, saliva, tears, feces, semen, mucous, tissue, tissue homogenate, cellular extract, or spinal fluid, and individual microspheres are conjugated to Zika virus (ZIKV) peptides that differ from ZIKV peptides to which other individual microspheres are conjugated, and the ZIKV peptides are selected from the group consisting of full-length ZIKV NS1, full-length ZIKV NS5, and full-length ZIKV envelope protein;
• forming a first incubated suspension wherein forming comprises incubating said first portion with said first suspension, in the presence of between 6 M urea and 10 M urea, to permit high-avidity binding between anti-ZIKV NS1, anti-ZIKV NS5, and anti-ZIKV envelope protein antibodies present in the first portion and microspheres of said first plurality of microspheres;
• forming a second incubated suspension wherein forming comprises contacting said first plurality of microspheres with an anti-ZIKV antibody detecting-reagent to permit binding between the anti-ZIKV antibody detecting reagent and antibodies bound to microspheres of said first plurality of microspheres;
• removing from the second incubated suspension anti-ZIKV antibody detecting-reagent molecules that are not bound to antibodies bound to microspheres;
• and
• detecting anti-ZIKV antibody detecting-reagent molecules in the second incubated suspension.