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A rapid visualization method for detecting rotavirus A by combining nuclear acid sequence-based amplification with
Yue Chen1, Junhua Wu2, E-Bin Gao3
1Health Science Center, Ningbo University, Ningbo, Zhejiang 315000, PR China.
Insights
A new NASBA-Cas12a system rapidly detects Rotavirus A, a common cause of severe childhood diarrhea. This nucleic acid detection method offers high sensitivity and specificity for early diagnosis and treatment.
Area of Science:
- Molecular Biology
- Virology
- Diagnostic Technology
Background:
- Rotavirus A is a leading cause of severe diarrhea in children under five, leading to serious health complications.
- Current diagnostic methods lack the speed and efficiency needed for timely intervention and transmission control.
Purpose of the Study:
- To develop a rapid and sensitive nucleic acid detection system for Rotavirus A.
- To utilize a combination of NASBA and Cas12a for enhanced diagnostic capabilities.
Main Methods:
- Development of a Nuclear Acid Sequence-Based Amplification (NASBA)-Cas12a system.
- Detection of Rotavirus A using fluorescence intensity or lateral flow strips.
Main Results:
- The NASBA-Cas12a system detected Rotavirus A within 70 minutes at 37°C with a limit of detection of 1.2 copies/μl.
- Achieved 100% sensitivity, 90% specificity, and high consistency with RT-PCR (kappa=0.933).
- Demonstrated no cross-reactivity with other common viruses.
Conclusions:
- The NASBA-Cas12a system is a highly sensitive and specific tool for Rotavirus A detection.
- This system shows significant potential for clinical applications in early diagnosis and management.
Abstract:
Introduction. Rotavirus A is the most common pathogen causing diarrhoea in children less than 5 years, leading to severe complications such as dehydration, electrolyte imbalances, acidosis, myocarditis, convulsions, pneumonia, and other life-threatening conditions.Gap statement. There is an urgent need for a rapid and efficient nucleic acid detection strategy to enable early diagnosis and treatment, preventing rotavirus transmission and associated complications.Aim. This article aimed to develop a nuclear acid sequence-based amplification (NASBA)-Cas12a system for detecting rotavirus A using fluorescence intensity or lateral flow strips.Methodology. The NASBA technology was combined with the clustered regularly interspaced short palindromic repeats-Cas12a system to establish a NASBA-Cas12a system for detecting rotavirus A.Results. The NASBA-Cas12a system could detect rotavirus A at 37 ℃ within 70 min and had no cross-reactivity with other viruses, achieving a limit of detection of 1.2 copies μl-1. This system demonstrated a sensitivity of 100%, specificity of 90%, positive predictive value of 97.22% and negative predictive value of 100%. The kappa value was 0.933, indicating that the NASBA-Cas12a system was highly consistent with reverse transcription-PCR.Conclusion. The NASBA-Cas12a system exhibited high sensitivity and specificity for detecting rotavirus A, showing great potential for clinical application.
