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Updated: Oct 21, 2025
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Rapid Internal Control Reference Recombinase-Aided Amplification Assays for EBV and CMV Detection
Yuan Gao1, Yan Qing Tie2, Lin Qing Zhao3
1Hebei Medical University, Shijiazhuang 050031, Hebei, China;NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;Hebei General Hospital, Shijiazhuang 050070, Hebei, China.
A new internal control reference recombinase-aided amplification (ICR-RAA) assay rapidly detects Epstein-Barr virus (EBV) and cytomegalovirus (CMV) within 30 minutes. This highly sensitive and specific assay shows promise for quick diagnosis of these common human herpesviruses.
Area of Science:
- Molecular Biology
- Virology
- Diagnostic Assays
Background:
- Epstein-Barr virus (EBV) and cytomegalovirus (CMV) are prevalent human herpesviruses linked to various diseases.
- Accurate and rapid detection of EBV and CMV is crucial for timely patient management.
Discussion:
- The developed internal control reference recombinase-aided amplification (ICR-RAA) assay provides detection within 30 minutes.
- The assay demonstrates high sensitivity (5 copies/test for EBV, 1 copy/test for CMV) and specificity, with no cross-reactivity.
- ICR-RAA performance was comparable to commercial quantitative polymerase chain reaction (qPCR), showing high sensitivity and specificity for both extracted DNA and thermally lysed samples.
Key Insights:
- The ICR-RAA assay achieves rapid (30-minute) and specific detection of EBV and CMV.
- Sensitivity and specificity of the ICR-RAA assay are comparable to established qPCR methods.
- The assay exhibits excellent agreement with qPCR, indicated by high Kappa values (0.930 for EBV, 0.892 for CMV with extracted DNA).
Outlook:
- The ICR-RAA assay offers a valuable tool for rapid clinical diagnostics of EBV and CMV infections.
- Further validation in diverse clinical settings can confirm its utility in routine diagnostics.
- This assay could potentially streamline the diagnostic workflow for herpesvirus infections.
