Method and kit for detection of anti-Zika virus antibodies
View Patent ↗Provided is a method of detecting the presence of an anti-Zika virus (ZIKV) antibody in a sample, including contacting a sample with a suspension having a plurality of microspheres wherein individual microspheres are conjugated to a peptide and the peptide includes a ZIKV peptide selected from the group including ZIKV NS1, ZIKV NS5, and ZIKV envelope protein, forming a first incubated suspension by incubating said sample with said suspension to permit binding of anti-ZIKV antibodies present in the sample to said microspheres, forming a second incubated suspension by contacting said first incubated suspension with an anti-ZIKV antibody detecting-reagent to permit binding of the anti-ZIKV antibody detecting reagent to said microspheres, removing from the second incubated suspension anti-ZIKV antibody detecting-reagent molecules that are not bound to said microspheres, and detecting the presence of anti-ZIKV antibody detecting-reagent molecules in the second incubated suspension. Also provided is a kit containing reagents and compositions for performing the foregoing method.
1. A method, comprising:
(i) incubating
(a) a first sample from a subject with a first plurality of populations of microspheres conjugated to viral antigens under a first condition, wherein
the viral antigens conjugated to microspheres of each population of the first plurality of populations of microspheres differ from the viral antigens conjugated to microspheres of all other populations of microspheres of the first plurality of populations of microspheres, and
the first condition permits high-affinity binding and low-affinity binding of antibodies in the first sample to the viral antigens, and
(b) a second sample from the subject with a second plurality of populations of microspheres conjugated to viral antigens under a second condition, wherein
the viral antigens conjugated to microspheres of each population of the second plurality of populations of microspheres differ from the viral antigens conjugated to microspheres of all other populations of microspheres of the second plurality of populations of microspheres, and
the second condition permits high-affinity binding but not low-affinity binding of antibodies in the second sample to the viral antigens, and
(ii) detecting an amount of antibodies bound to each population of the first plurality of populations of microspheres and to each population of the second plurality of populations of microspheres, wherein
a first viral antigen comprises full-length Zika virus (ZIKV) NS1, a second viral antigen comprises full-length ZIKV envelope protein, and a third viral antigen comprises a Dengue virus (DENV) antigen wherein the DENV antigen is selected from DENV1 NS1, DENV2 NS1, DENV3 NS1, and DENV4 NS1.
2. The method of claim 1 , wherein a fourth viral antigen is full-length ZIKV NS5.
3. The method of claim 1 , wherein the second condition comprises incubating in the presence of from 7 M urea to 9 M urea.
4. The method of claim 1 , wherein the second condition comprises incubating in the presence of 8 M urea.
5. The method of claim 2 , wherein the second condition comprises incubating in the presence of from 7 M urea to 9 M urea.
6. The method of claim 2 , wherein the second condition comprises incubating in the presence of 8 M urea.
7. The method of claim 1 , wherein the first sample, the second sample, or both samples comprise blood, serum, plasma, saliva, tears, feces, semen, mucous, tissue, tissue homogenate, cellular extract, and spinal fluid.
8. The method of claim 2 , wherein the first sample, the second sample, or both samples comprise blood, serum, plasma, saliva, tears, feces, semen, mucous, tissue, tissue homogenate, cellular extract, and spinal fluid.
9. The method of claim 1 , wherein first sample and the second sample comprise serum.
10. The method of claim 2 , wherein first sample and the second sample comprise serum.