Anti-Hepatitis A-Virus (HAV) IgG ELISA Kit

Sample

Serum and plasma

Species Reactivity

Human

Detection Method

cELISA

Intended Use

Anti-HAV IgG ELISA Kit is an enzyme immunoassay for the detection of antibodies against Hepatitis A virus in Human Serum for research use.

Contents of Kit

1. Microtiter plate: ready for use, coated with anti-Mouse Adiponectin antibody. Wells are separately breakable. (8x12) wells
2. Conjugate Concentrate (KK): 100-fold concentrated (peroxidase labeled Anti-HAV Ig). 1 x 100 μL
3. Positive Control (PK): ready for use, anti-HAV positive control serum >500 mIU/mL. 1 x 1 mL
4. Negative Control (NK): ready for use, anti-HAV negative control serum. 1 x 1 mL
5. Dilution Buffer (VP): ready for use. Please shake before use! 1 x 120 mL
6. Washing Buffer (WP): 20-fold concentrated solution. 1 x 50 mL
7. Substrate (S): ready for use, horseradish-peroxidase-(HRP) substrate. 1 x 12 mL
8. Stopping Solution (SL): ready for use, 0.2 M sulfuric acid. 1 x 12 mL
9. Sealing Tape: for covering the microtiter plate. 2
Additional to Semi-Quantitative Test:
10. Serum standard 1 (STD1): ready for use, anti-HAV titer 50 mIU/mL. 1 x 1 mL
11. Serum standard 2 (STD2): ready for use, anti-HAV titer 30 mIU/mL. 1 x 1 mL
12. Serum standard 3 (STD3): ready for use, anti-HAV titer 10 mIU/mL. 1 x 1 mL

Storage

Store the kit at 2-8°C after receipt until its expiry date. Avoid repeated thawing and freezing.
The shelf life of the components after initial opening is warranted for 4 weeks, store the unused strips and microtiter wells airtight together with the desiccant at 2-8°C in the clip-lock bag, use in the frame provided. The 1:20 diluted Washing Buffer WP is 4 weeks stable at 2-8°C

Precision

Serum samples were measured in at least 13 independent assays and the variation of the measurement result was calculated (Table 4).
Table 4: Inter-Assay Variation. SD= Standard deviation, CV% = Coefficient of Variation in %, Number = Number of the independent determinations.

Sensitivity

The sensitivity of the test system was evaluated by the International standard NIBSC 97/646. The standard material was diluted and the dilution was applied as sample in the Anti-HAV IgG ELISA Kit. The measured signals (OD450) are shown in table 5.
Table 5: Analytical sensitivity. Shown are the measured signals (OD450).

The recalculated analytical sensitivity resulting from the 2fold standard deviation of the negative control (n=6) is 1.10 mIU/mL. In an additional experiment the theoretically calculated analytical sensitivity was proven by 1 mIU/mL diluted NIBSC 97/646. Here the negative control showed a signal of 2.5 (SD 0.18) and the diluted international standard had a signal of 2.12 (SD 0.2).
Furthermore, through the measurement of 801 human serum samples (of which 287 were positive and 514 negative for anti-HAV antibodies) can be shown that the Anti-HAV IgG ELISA Kit distinguishs very well between positive and negative samples, the sensitivity was 98.26 %.

General Description

After infection with the Hepatitis A-Virus, neutralising antibodies appear at the same time of Anti_x0002_HAV of IgG-Class formation. The Anti-HAV titers 3 to 6 months after naturally occurring infections are very high, within the range of 100 to more than 300 International Units per ml (IU/ml). Even after more than 10 years the titers usually remain at more than 1 to 10 IU/ml. A value of 10 - 30 (mIU/ml) milli International Units can be considered the minimal protective level. Since the incidence of HAV infection in children has diminished in Northern Europe in recent years, children and juveniles are predominantly Anti-HAV negative. In an adult population could be observed, that the percentage of anti-HAV positive individuals raises with increasing age: only 3.9 % of the young people (18-24 years) show antibody against HAV, where as in older population (≥50 years) HAV-Antibody was found in 40.3 % of the population. In countries with less favourable sanitary conditions around the Mediterranean area, Africa or Asia, the incidence is very high. Higher risk is also given for clinical staff, especially in the paediatric. Since a vaccine against Hepatitis A virus infections is available, vaccinations are recommended for people travelling to countries where a high risk of HAV infections exists and for health care employees.