Monkeypox PCR controls 

Monkeypox PCR controls are essential in the molecular detection and quantification of the Monkeypox virus (MPXV), an Orthopoxvirus responsible for the monkeypox disease. These controls ensure the accuracy, sensitivity, and specificity of PCR assays used in detecting MPXV DNA in clinical and environmental samples, especially during outbreaks.

Content of Monkeypox PCR Controls

Positive Control

• Contains a known concentration of MPXV DNA.
• Ensures that the PCR assay can correctly identify the presence of MPXV DNA in a sample.
• Verifies the proper functioning of all PCR components, including primers, probes, thermal cycling conditions, and reagents, ensuring the detection of true positives.

Negative Control

• Includes all PCR reagents without MPXV DNA.
• Used to confirm that there is no contamination in the PCR reagents or laboratory environment, preventing false-positive results.
• Ensures assay specificity by verifying that no non-specific amplification or cross-reactivity with non-target sequences (e.g., other Orthopoxviruses) occurs.

Internal Control

• A non-target DNA sequence, either synthetic or endogenous, is added to each reaction.
• Confirms that no PCR inhibitors are present in the sample that could interfere with the amplification process.
• Helps differentiate between true negatives (no MPXV DNA present) and false negatives (MPXV DNA present but not detected due to inhibition), maintaining the reliability of the results.

Reference Control

• A well-characterized sample with a known amount of MPXV DNA.
• Used to assess the quantitative performance of the PCR assay, such as determining the limit of detection (LOD) and ensuring consistency across different batches and assay runs.
• Provides a benchmark for interpreting quantitative results, crucial for understanding viral load dynamics during infection and monitoring the effectiveness of treatments or public health interventions.