**Human HPV IgG ELISA Kit – For the Quantitative In Vitro Determination of Human Parvovirus IgG in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Body Fluids**
**For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.**
Before using this product, please read the complete package insert thoroughly. This kit is designed for research purposes only and should not be used in clinical diagnostic settings.
**Intended Use and Test Principle**
The Human HPV IgG ELISA Kit is intended for laboratory research use only and is not suitable for diagnostic or therapeutic procedures. The test is based on a competitive enzyme-linked immunosorbent assay (ELISA) principle. The reaction is terminated by a Stop Solution, which changes the color from blue to yellow. The optical density (OD) is measured at 450 nm using a microplate reader.
A set of calibration standards is included in the kit to generate a standard curve. By comparing the OD values of the samples with those of the standards, the concentration of HPV IgG in the samples can be accurately determined.
**Storage and Handling Instructions**
- Store all samples at -20°C. Avoid repeated freeze-thaw cycles.
- For cell culture supernatants, tissue homogenates, and other biological fluids, remove particulates by centrifugation and assay immediately or aliquot and store at -20°C.
- Ensure proper centrifugation and avoid hemolysis or contamination.
**Materials Required but Not Supplied**
1. 37°C incubator
2. Microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
**Reagents Provided**
All reagents are stored at 2–8°C. Check the expiration date on the label before use.
| Reagent Name | 96 Determinations | 48 Determinations |
|----------------------------------|-------------------|-------------------|
| MicroELISA Strip Plate | 12×8 strips | 12×4 strips |
| Standard (6 vials) | 0.5 ml/vial | 0.5 ml/vial |
| Sample Diluent | 6.0 ml | 3.0 ml |
| HRP-Conjugate Reagent | 10.0 ml | 5.0 ml |
| 20X Wash Solution | 25 ml | 15 ml |
| Chromogen Solution A | 6.0 ml | 3.0 ml |
| Chromogen Solution B | 6.0 ml | 3.0 ml |
| Stop Solution | 6.0 ml | 3.0 ml |
| Closure Plate Membrane | 2 | 2 |
| User Manual | 1 | 1 |
| Sealed Bags | 1 | 1 |
**Important Notes**
- Standard concentrations: 80, 40, 20, 10, 5, 2.5 U/L
- If sample values exceed the highest standard, dilute with Sample Diluent.
- Always use reagents provided by the manufacturer.
**General Precautions**
1. Allow all reagents and materials to reach room temperature (20–25°C) before use. Do not thaw using water baths.
2. Do not use reagents beyond their expiration date.
3. Use only deionized or distilled water for dilutions.
4. Keep microtiter plates in sealed bags until needed. Store unused strips at 2–8°C with desiccant.
5. Use fresh pipette tips for each transfer to prevent cross-contamination.
6. Treat all disposable items as potentially hazardous. Follow proper biosafety protocols.
**Waste Disposal Guidelines**
- For liquid waste: Add sodium hypochlorite to achieve a final concentration of 1.0%. Allow to stand for 30 minutes to inactivate viruses before disposal.
- Substrate Solution A and B are sensitive to contamination.
- Substrate B contains 20% acetone; keep away from heat or open flame.
**Reagent Preparation and Storage**
- **Wash Solution (1X):** Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to 1 month.
**Assay Procedure**
1. Prepare all reagents before starting. Run standards and samples in duplicate.
2. Add 50 µL of standard or sample to appropriate wells. Blank well receives no addition.
3. Add 100 µL of HRP-conjugate reagent to all wells except the blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the plate 4 times manually or using an automated washer.
5. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
6. Add 50 µL of Stop Solution to each well. Read OD at 450 nm.
**Data Analysis**
- Plot average OD values of standards against their concentrations to generate a standard curve.
- Subtract blank OD from all measurements.
- Locate the OD value on the Y-axis and draw a horizontal line to intersect the standard curve. Drop a vertical line to the X-axis to determine the concentration.
- Intra-assay and inter-assay CV% are <15%.
- Assay range: 2.5 – 80 U/L.
- Sensitivity: <1.0 U/L.
- Cross-reactivity: No significant interference observed.
**Storage Conditions**
- Store at 2–8°C for frequent use.
- For long-term storage, keep at -20°C for up to six months.
**Important Reminder**
This kit must be used strictly according to the instructions. Any deviation may affect accuracy and reliability. Always follow good laboratory practices and safety protocols.
**Please read this instruction carefully before proceeding.**
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