DESCRIPTION

Used for the quantitative determination of Human Anti-Monkeypox virus IgG (M1R) concentration in serum and plasma.

PRINCIPLE OF THE ASSAY

This assay employs the quantitative competitive enzyme immunoassay technique. Recombinant Monkeypox virus M1R has been pre-coated onto a microplate. Standards or samples are premixed with biotin-labeled antibody and then pipetted into the wells. Human Anti-Monkeypox virus IgG (M1R) in the sample competitively binds to the pre-coated protein with biotin-labeled Human Anti-Monkeypox virus IgG (M1R). After washing away any unbound substances, Streptavidin-HRP is added to the wells. Following a wash to remove any unbound enzyme reagent, a substrate solution is added to the wells and color develops in inversely proportion to the amount of Human Anti-Monkeypox virus IgG (M1R) bound in the initial step. The color development is stopped and the intensity of the color is measured.

PRECISION

Intra-Assay Precision (Precision within an assay): <20% Three samples of known concentration were tested sixteen times on one plate to assess intra-assay precision. Inter-Assay Precision (Precision between assays): <20% Three samples of known concentration were tested in twenty four separate assays to assess inter-assay precision.

Sample		1	2	3	1	2	3
n		16	16	16	24	24	24
Mean (ng/mL)		377.5	110.5	32.0	360.5	102.8	32.4
Standard deviation		13.3	4.7	4.3	21.4	3.7	3.3
CV \(%\)		3.5	4.2	13.6	5.9	3.6	10.1

RECOVERY

The recovery of Human Anti-Monkeypox virus IgG (M1R) spiked to three different levels in human serum samples throughout the range of the assay was evaluated.

Sample	Dilution factor	Average (%)	Range (%)
Human serum (n=4)	1:10	105	86-120
	1:20	105	87-122

LINEARITY

To assess the linearity of the assay, human serum samples were spiked with high concentrations of Human Anti-Monkeypox virus IgG (M1R) and diluted with the Standard Diluent to produce samples with values within the dynamic range of the assay.

Sample	Dilution factor	Average (%)	Range (%)
Human serum (n=4)	1:2	87	81-93
	1:4	90	83-100
	1:8	95	89-105
	1:16	96	90-109