Wang Kun, Ai Yuanhang, Luo Juan, Liang Longying, Zhang Weiwei, Cao Guojun, Zha He, Wu Jie, Lei Kun, Yao Shifei, Wu Kaifeng
Department of Laboratory Medicine, The Third Affiliated Hospital of Zunyi Medical University (The First People's Hospital of Zunyi), Guizhou, China.
Department of Pediatrics and Child Health, The Third Affiliated Hospital of Zunyi Medical University (The First People's Hospital of Zunyi), Guizhou, China.
Front Cell Infect Microbiol. 2025 May 13;15:1563495. doi: 10.3389/fcimb.2025.1563495. eCollection 2025.
Coxsackievirus A6 (CVA6) ranks as a primary enterovirus associated with hand-foot-mouth disease (HFMD) and herpangina (HA). Given its significant role in these diseases, there is an urgent need for an efficient identification method. This study presents a novel visual approach based on the reverse transcription polymerase spiral reaction (RT-PSR) for the rapid detection of CVA6. We designed an RT-PSR assay that targets and amplifies a segment of the VP1 gene. Hydroxy naphthol blue (HNB) is incorporated as the detection agent in this assay. To evaluate the performance of the RT-PSR assay, we analyzed 142 clinical throat swab samples. The results were benchmarked against those obtained using quantitative reverse transcription - polymerase chain reaction (qRT - PCR). The RT-PSR assay operates at 65°C for 60 minutes and exhibits a detection limit of 10 copies/μL. When tested against other viruses, it consistently yielded negative results, demonstrating its high specificity. Moreover, the RT - PSR assay showed excellent agreement with a commercial qRT - PCR kit. In conclusion, by using HNB as an indicator, the RT - PSR assay emerges as a straightforward and highly sensitive method for detecting CVA6 in symptomatic throat samples. This approach holds great potential for improving the diagnosis and surveillance of CVA6 - related diseases.
柯萨奇病毒A6（CVA6）是与手足口病（HFMD）和疱疹性咽峡炎（HA）相关的主要肠道病毒。鉴于其在这些疾病中的重要作用，迫切需要一种高效的鉴定方法。本研究提出了一种基于逆转录聚合酶螺旋反应（RT-PSR）的新型可视化方法，用于快速检测CVA6。我们设计了一种针对并扩增VP1基因片段的RT-PSR检测方法。在该检测方法中加入了羟基萘酚蓝（HNB）作为检测试剂。为了评估RT-PSR检测方法的性能，我们分析了142份临床咽拭子样本。将结果与使用定量逆转录-聚合酶链反应（qRT-PCR）获得的结果进行了对比。RT-PSR检测方法在65°C下运行60分钟，检测限为10拷贝/μL。当针对其他病毒进行检测时，它始终产生阴性结果，表明其具有高特异性。此外，RT-PSR检测方法与商用qRT-PCR试剂盒显示出极好的一致性。总之，通过使用HNB作为指示剂，RT-PSR检测方法成为一种用于检测有症状咽拭子样本中CVA6的简单且高度灵敏的方法。这种方法在改善CVA6相关疾病的诊断和监测方面具有巨大潜力。
