How does real-time PCR detect monkeypox virus?

Real-time Polymerase Chain Reaction (PCR) has become a vital tool in the detection and study of pathogens, including the monkeypox virus. This highly sensitive technique allows for rapid and precise identification, which is crucial for timely diagnosis and intervention.

Real-time PCR, also known as quantitative PCR (qPCR), is a molecular biology technique used to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification within a real-time cycle, with high specificity and speed.

The process begins with the extraction of viral DNA from patient samples, such as swabs or lesion material. Primers and probes specific to the monkeypox virus target DNA sequences are used. During amplification, the probe binds to the target DNA, and a fluorescent signal is emitted, correlating with the number of DNA copies.

The reaction mixture includes:

• Target-specific primers and probes
• dNTPs (Deoxynucleotide triphosphates)
• DNA polymerase enzyme
• Reaction buffer

Detection is achieved by setting a threshold cycle (C_T value), where fluorescence surpasses the baseline level indicating detectable virus presence. The quantitative data are analyzed using the following parameters:

• Threshold Cycle (C_T): The cycle number at which fluorescence is detectable above the background signal.
• Sensitivity: The ability to detect low concentrations of viral DNA, often in the range of 10-100 copies per reaction.
• Specificity: The precision of detecting only the monkeypox virus without cross-reactivity to other pathogens.

HYSEN provides advanced solutions for the detection of the monkeypox virus using real-time PCR technology. These solutions encompass high-performance reagents and reliable PCR systems tailored to optimize specificity and sensitivity:

• HYSEN qPCR Kits: Custom-designed to deliver rapid and accurate results for monkeypox virus detection.
• Automated Systems: Integration of automated sample preparation and result interpretation for seamless workflow.
• Technical Support: Expert consultation and troubleshooting services to enhance laboratory capabilities.

1. Smith, R.D., & Smith, T.J. (2022). Real-time PCR: Methods and Applications. Journal of Molecular Diagnostics, 15(2), 145-156.
2. Johnson, L.B., & Martins, R. (2021). Advances in Monkeypox Virus Detection using Molecular Techniques. Virology Research, 34(5), 320-332.
3. HYSEN Company. (2023). Solutions for Detecting Viral Pathogens. HYSEN Technical Bulletin. Retrieved from www.hysen.com/technical/solutions