Evaluation of the MonkeyPox AffiTest: A Rapid Lateral Flow Immunoassay for Detection of MPXV Antigens

MonkeyPox AffiTest is an immunoassay designed for the rapid detection of monkeypox virus (MPXV) antigens in clinical samples. This article provides a technical evaluation of the AffiTest, focusing on its methodological principles, performance characteristics, and clinical utility.

Monkeypox is a zoonotic viral infection caused by the monkeypox virus (MPXV), a member of the Orthopoxvirus genus. Accurate and timely diagnosis is critical for managing outbreaks and implementing appropriate public health measures. The MonkeyPox AffiTest is a lateral flow immunoassay intended for the qualitative detection of MPXV antigens in patient specimens.

Principle of the Assay

The MonkeyPox AffiTest operates on the principle of lateral flow immunoassay technology. It utilizes specific antibodies that bind to MPXV antigens present in the sample. The test strip consists of several key components:

  • Sample Pad: Where the specimen is applied.
  • Conjugate Pad: Contains antibodies conjugated with gold nanoparticles, which bind to MPXV antigens.
  • Nitrocellulose Membrane: Contains test and control lines where antigen-antibody complexes are captured.
  • Absorbent Pad: Facilitates the flow of reagents through the test strip.

When the sample is applied, it migrates through the conjugate pad, where MPXV antigens bind to the conjugated antibodies. The antigen-antibody complexes then travel along the nitrocellulose membrane and are captured at the test line, resulting in a visible signal if MPXV is present.

Sample Collection

The test requires clinical specimens such as skin lesions, vesicular fluid, or swabs from affected areas. Samples are prepared according to the manufacturer’s instructions to ensure accuracy and reliability.

Procedure

  • Apply the specimen to the sample pad.
  • Allow the sample to migrate through the conjugate pad and the nitrocellulose membrane.
  • Read the result within the specified time frame (typically 15-20 minutes) to avoid false results.

Performance Characteristics

Sensitivity and Specificity

The sensitivity of the MonkeyPox AffiTest is determined by its ability to correctly identify positive cases of monkeypox, while specificity refers to its ability to correctly identify negative cases. Performance metrics are evaluated through clinical validation studies:

  • Sensitivity: Typically >90% (dependent on study conditions and sample types).
  • Specificity: Typically >95% (dependent on cross-reactivity with other pathogens).

Limit of Detection (LoD)

The LoD indicates the smallest amount of MPXV antigen detectable by the test. The AffiTest’s LoD is calibrated to detect clinically relevant levels of MPXV antigen to ensure effective diagnosis during early stages of infection.

Stability and Storage

The test should be stored at 2-30°C and protected from direct sunlight. Stability studies confirm that the test maintains its performance characteristics within the recommended storage conditions and up to the expiration date.

Clinical Utility​

The MonkeyPox AffiTest offers rapid results, making it suitable for point-of-care settings and field diagnostics. It complements traditional diagnostic methods such as PCR by providing quick preliminary results, aiding in the early detection and management of monkeypox cases.

Limitations

While the MonkeyPox AffiTest is highly effective, it has limitations:

  • False Negatives: Can occur if the antigen levels are below the test’s LoD.
  • False Positives: May arise due to cross-reactivity with other orthopoxviruses or similar pathogens.

The MonkeyPox AffiTest is a valuable tool for the rapid detection of monkeypox virus antigens. Its technical design ensures ease of use and quick results, which are essential for timely disease management and control. Ongoing evaluation and quality control are necessary to maintain its diagnostic accuracy and reliability.

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