Enterovirus (EV) PCR controls are essential components in the molecular detection of Enteroviruses, a group of RNA viruses that include polioviruses, coxsackieviruses, echoviruses, and others. These controls ensure the accuracy, sensitivity, and specificity of PCR assays used in detecting EV RNA in clinical and environmental samples.

Content of EV PCR Controls

Positive Control

• Contains a known concentration of EV RNA.
• Ensures the PCR assay can correctly identify the presence of EV RNA in the sample.
• Verifies that the reverse transcription process (if using RT-PCR), primers, probes, thermal cycling conditions, and reagents are functioning properly, ensuring true positive results.

Negative Control

• Includes all PCR reagents without EV RNA.
• Used to verify that there is no contamination in the PCR reagents or laboratory environment that could lead to false-positive results.
• Ensures the specificity of the assay by confirming that no non-specific amplification or cross-reactivity with non-target sequences occurs.

Internal Control

• A non-target RNA sequence is added to each reaction.
• Confirms that PCR inhibitors present in the sample do not interfere with the amplification process.
• Helps distinguish between true negatives (no EV RNA present) and false negatives (EV RNA present but not detected due to PCR inhibition), ensuring reliable and accurate results.

Reference Control

• A well-characterized sample with a known quantity of EV RNA.
• Used to assess the quantitative performance of the PCR assay, such as determining the limit of detection (LOD) and ensuring consistency across different assay runs and reagent batches.
• Provides a benchmark for interpreting results, ensuring the reproducibility and reliability of the assay, particularly in clinical diagnostics and public health monitoring.