DESCRIPTION

Used for the quantitative determination of HSV-1 gD concentration in serum and plasma.

PRINCIPLE OF THE ASSAY

This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for HSV-1 gD has been pre-coated onto a microplate. Standards or samples are pipetted into the wells and any HSV-1 gD present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-labeled antibody specific for HSV-1 gD is added to the wells. 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 proportion to the amount of HSV-1 gD 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): <10% Three samples of known concentration were tested sixteen times on one plate to assess intra-assay precision. Inter-Assay Precision (Precision between assays): <15% 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)		18.6	3.4	1.4	18.8	3.6	1.4
Standard deviation		0.9	0.1	0.1	0.9	0.2	0.1
CV \(%\)		4.8	4.1	9.4	4.7	6.3	9.4

RECOVERY

The recovery of HSV-1 gD 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=3)	1:10	96	81-114
	1:20	92	84-102

LINEARITY

To assess the linearity of the assay, human serum samples were spiked with high concentrations of HSV-1 gD 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=3)	1:2	110	103-116
	1:4	111	100-119
	1:8	106	94-116
	1:16	98	87-107