MonkeyPox Virus IgG (MPV-IgG) ELISA Kit

The MonkeyPox Virus IgG (MPV-IgG) ELISA Kit is a qualitative enzyme-linked immunosorbent assay designed for the detection of Monkey MPV-IgG antibodies in serum, plasma, culture media, and other biological fluids.

This microplate-based assay utilizes antigen-coated wells and an HRP–TMB colorimetric detection system to deliver reliable and reproducible results measured at 450 nm. The kit is suitable for research laboratories conducting immunological, virology, and infectious disease studies.

This product is for research use only and is not intended for therapeutic or diagnostic applications.

Intended Use

Qualitative detection of Monkey MPV-IgG antibodies in:

• Serum
• Plasma
• Culture media
• Urine and cerebrospinal fluid
• Cell culture supernatants
• Tissue homogenates

Assay Principle

The assay is based on the ELISA technique:

1. Microplate wells are pre-coated with Monkey MPV-specific antigen.
2. IgG antibodies in the sample bind to the immobilized antigen during incubation.
3. An HRP-labeled antigen conjugate binds to the captured antibodies.
4. After washing, TMB substrate is added, producing a blue color reaction.
5. Addition of stop solution converts the reaction to yellow.
6. Optical density (OD) is measured at 450 nm.
7. Results are interpreted relative to a calculated cut-off value.

The intensity of the color is proportional to the amount of MPV-IgG present in the sample.

Kit Components

The kit allows 48 or 96 reactions, including blank, negative, and positive controls.

Required Equipment (Not Provided)

• ELISA microplate reader (450 nm)
• 37 °C incubator
• Manual or automatic plate washer
• Adjustable pipettes (10–1000 µL)
• Vortex mixer
• Distilled or deionized water
• Timer

Sample Preparation

Serum:

Allow blood to clot at room temperature for 10–20 minutes. Centrifuge at 2,000–3,000 rpm for 20 minutes and collect serum.

Plasma:

Collect blood in EDTA or citrate tubes. Centrifuge at 2,000–3,000 rpm for 20 minutes and collect plasma.

Urine and Other Fluids:

Centrifuge at 2,000–3,000 rpm for 20 minutes and collect clear supernatant.

Cell Samples:

Centrifuge culture supernatants and collect clarified supernatant. For intracellular detection, lyse cells in PBS (pH 7.2–7.4) via freeze–thaw cycles and centrifuge.

Tissue Samples:

Homogenize tissue in PBS (pH 7.4), centrifuge at 2,000–3,000 rpm for 20 minutes, and collect supernatant.

Important Notes:

• Store samples at −20 °C if testing is delayed.
• Avoid repeated freeze–thaw cycles.
• Samples containing sodium azide (NaN₃) are incompatible due to inhibition of HRP activity.

Assay Procedure Summary

1. Add controls and diluted samples to designated wells.
2. Incubate at 37 °C for 30 minutes.
3. Wash plate five times with 1X wash buffer.
4. Add HRP-conjugate reagent and incubate at 37 °C for 30 minutes.
5. Wash plate five times.
6. Add Chromogen Solutions A and B; incubate at 37 °C for 15 minutes in the dark.
7. Add Stop Solution.
8. Measure absorbance at 450 nm within 15 minutes.

Thorough washing is critical to ensure assay precision and minimize background.

Result Interpretation

Validation Criteria:

• Average OD of Positive Control ≥ 1.00
• Average OD of Negative Control ≤ 0.10

Cut-Off Calculation:

Cut-Off = Average Negative Control + 0.15

Interpretation:

• OD < Cut-Off: Negative
• OD ≥ Cut-Off: Positive

Storage and Stability

• Store unopened kit at 2–8 °C.
• Shelf life: 6 months.
• After opening, reseal unused strips with desiccant and store at 2–8 °C.
• Use opened strips within 1 month (maximum 3 months).

The MonkeyPox Virus IgG (MPV-IgG) ELISA Kit provides a reliable and standardized qualitative method for detecting MPV-specific IgG antibodies in a wide range of biological samples. Its pre-coated microplate format, validated controls, and optimized HRP–TMB detection system ensure reproducible results suitable for research laboratories in virology, immunology, and infectious disease studies.