Rotavirus Antigen ELISA Kit

Name: Rotavirus Antigen ELISA Kit
SKU: DEIAFY102
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

stool samples

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Rotavirus assay is an enzyme-linked immunosorbent assay (ELISA) for qualitative identification of rotaviruses in human stool samples. For Research Use Only. Not for use in diagnostic procedures.

Contents of Kit

No.	Components	Size	Storage Conditions
1	Plate	96 wells, 12 microwell strips (can be divided) in the strip holder	2-8°C
2	Positive Control	2 mL	2-8°C
3	Negative Control	2 mL	2-8°C
4	Diluent 1	100 mL	2-8°C
5	Conjugate 1	13 mL	2-8°C
6	Conjugate 2	13 mL	2-8°C
7	Substrate 1	3 mL	2-8°C
8	Wash Buffer 10×	100 mL	2-8°C
9	Stop solution	12 mL	2-8°C
10	Plate Seater	2

Storage

All reagents must be stored at 2 - 8°C and can be used until the expiry date printed on the label. The 1× working wash buffer can be used for a maximum of 4 weeks when stored at 2 - 8°C. Microbial contamination must be prevented. After the expiry date, the quality guarantee is no longer valid.
The aluminum bag containing the microplate strips must be opened in such a way that the clip seal is not torn off. Any microplate strips which are not required must immediately be returned to the aluminum bag with the desiccant and stored at 2 - 8°C until all the strips have been used.
The colorless substrate must also be protected from direct light to prevent it from decomposing or turning blue due to auto-oxidation. Once the substrate has turned blue, it must not be used.

Performance Characteristics

The Rotavirus ELISA was validated in comparison with three commercial rotavirus ELISAs. The sample used consisted of fresh, same-day samples taken at a routine laboratory and of prepared samples frozen in advance at -20 °C for use in the comparison study. One homogeneous baseline suspension was tested by each of the ELISAs in accordance with the manufacturers' instructions.
A sample was considered positive or negative if the results of two out of three reference tests were in agreement. See the results of that study in Table.
Table: Correlation between our Rotavirus ELISA and three other commercial ELISAs

Sensitivity : 95,7 % Specificity : 99,1 %

Precision

Reproducibility of the the Rotavirus ELISA was tested with six references representing the complete measurement range from weak to high positive. To determine intra-assay reproducibility, 40 replicates of these references were assayed. Kit 1 was assayed with reference standard 1 (RS 1), and Kits 2 and 3 with reference standard 2 (RS 2). The mean values and coefficient of variation (CV) were determined for all three lots. For inter-assay reproducibility, references from 10 different working days were assayed in duplicate with two runs per day. The measurements were determined in three lots by three technicians. The inter-lot reproducibility was determined for two lots (Lots 2 and 3). See Table for results.
Table: Reproducibility and precision of the Rotavirus ELISA

Sensitivity

The analytical sensitivity of the Rotavirus ELISA was determined with the serial dilution of a stool sample quantified by immunoelectron microscopy (IEM). The measurements were taken in triplicate, based on a virus titer of 10.6 x 10⁷ particles/ml. The detection limit was defined as 6.63 × 10⁷ virus particles/ml of the stool sample. Results of the titration series are shown in Table.
Note that the positive OD value in ELISA is caused not only by intact virus particles, but also by fragments of the virus, which are not counted in the IEM.
Table: Determination of the analytical sensitivity of Rotavirus ELISA