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Background <p>As a major etiological agent of hand, foot, and mouth disease in humans, <i>Coxsackievirus A16</i> (CVA16) exerts a detrimental impact on the health of infants and toddlers. In the absence of effective antiviral drugs and vaccines targeting CVA16, a diagnostic strategy with sensitivity, specificity and rapid response is indispensable for the prevention and management of CVA16 infection.</p> Results <p>In this study, a novel molecular diagnostic approach, coupling real-time fluorescence technique with reverse transcription multiple cross-displacement amplification (RT-MCDA), was developed for the sensitive, rapid, and specific identification of CVA16 (termed CVA16-E-RT-MCDA). To construct the system, a unique set of CVA16-E-RT-MCDA primers was successfully designed targeting the VP1 gene of CVA16, and the optimal conditions for CVA16-E-RT-MCDA were determined to be 63 ℃ for 35&#xa0;min. The limit of detection for the CVA16-E-RT-MCDA assay was 14 copies/µL (approximately 70 copies per reaction) for the RNA standard template of CVA16. The CVA16-E-RT-MCDA assay exhibited high specificity and anti-interference. The findings demonstrated that the applicability of the CVA16-E-RT-MCDA method was promptly and successfully confirmed using clinical samples from patients with suspected CVA16 infections.</p> Conclusions <p>The CVA16-E-RT-MCDA assay established in the present study exhibits sensitivity, specificity and rapidity for CVA16 detection, rendering it a promising diagnostic tool for clinical applications.</p>

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Highly sensitive and rapid determination of Coxsackievirus A16 using restriction endonuclease‐mediated reverse transcription multiple cross displacement amplification

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月25日

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  • Qi Liang,
  • Yumei Cao,
  • Xujian Zhang,
  • Rui Ye,
  • Mao Liu,
  • Shu Zhang,
  • Yu Wang

摘要

Background

As a major etiological agent of hand, foot, and mouth disease in humans, Coxsackievirus A16 (CVA16) exerts a detrimental impact on the health of infants and toddlers. In the absence of effective antiviral drugs and vaccines targeting CVA16, a diagnostic strategy with sensitivity, specificity and rapid response is indispensable for the prevention and management of CVA16 infection.

Results

In this study, a novel molecular diagnostic approach, coupling real-time fluorescence technique with reverse transcription multiple cross-displacement amplification (RT-MCDA), was developed for the sensitive, rapid, and specific identification of CVA16 (termed CVA16-E-RT-MCDA). To construct the system, a unique set of CVA16-E-RT-MCDA primers was successfully designed targeting the VP1 gene of CVA16, and the optimal conditions for CVA16-E-RT-MCDA were determined to be 63 ℃ for 35 min. The limit of detection for the CVA16-E-RT-MCDA assay was 14 copies/µL (approximately 70 copies per reaction) for the RNA standard template of CVA16. The CVA16-E-RT-MCDA assay exhibited high specificity and anti-interference. The findings demonstrated that the applicability of the CVA16-E-RT-MCDA method was promptly and successfully confirmed using clinical samples from patients with suspected CVA16 infections.

Conclusions

The CVA16-E-RT-MCDA assay established in the present study exhibits sensitivity, specificity and rapidity for CVA16 detection, rendering it a promising diagnostic tool for clinical applications.