Zhong Zecheng, Su Xiaosong, Yang Kunyu, Huang Weida, Wang Jin, Zhuo Zhihao, Xiang Jiyu, Lin Lesi, He Shuizhen, Li Tingdong, Zhang Jun, Ge Shengxiang, Zhang Shiyin, Xia Ningshao
State Key Laboratory of Vaccines for Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, Fujian, China.
NMPA Key Laboratory for Research and Evaluation of Infectious Disease Diagnostic Technology, School of Public Health, Xiamen University, Xiamen, 361102, Fujian, China.
Nat Commun. 2024 Aug 1;15(1):6478. doi: 10.1038/s41467-024-50921-w.
Human enteroviruses (HEV) can cause a range of diseases from mild to potentially life-threatening. Identification and genotyping of HEV are crucial for disease management. Existing typing methods, however, have inherent limitations. Developing alternative methods to detect HEV with more virus types, high accuracy, and sensitivity in an accessible manner presents a technological and analytical challenge. Here, a sequence-specific nanoparticle barcode (SSNB) method is presented for simultaneous detection of 10 HEV types. This method significantly increases sensitivity, enhancing detection by 10-10 times over the traditional multiplex hybrid genotyping (MHG) method, by resolving cross-interference between the multiple primer sets. Furthermore, the SSNB method demonstrates a 100% specificity in accurately distinguishing between 10 different HEV types and other prevalent clinical viruses. In an analysis of 70 clinical throat swab samples, the SSNB method shows slightly higher detection rate for positive samples (50%) compared to the RT-PCR method (48.6%). Additionally, further assessment of the typing accuracy for samples identified as positive by SSNB using sequencing method reveals a concordance rate of 100%. The combined high sensitivity and specificity level of the methodology, together with the capability for multiple type analysis and compatibility with clinical workflow, make this approach a promising tool for clinical settings.
人肠道病毒（HEV）可引发一系列疾病，从轻度到可能危及生命不等。HEV的鉴定和基因分型对于疾病管理至关重要。然而，现有的分型方法存在固有局限性。开发能够以可及方式检测更多病毒类型、具备高精度和高灵敏度的HEV替代方法，是一项技术和分析上的挑战。在此，我们提出一种序列特异性纳米颗粒条形码（SSNB）方法，用于同时检测10种HEV类型。该方法显著提高了灵敏度，通过解决多个引物组之间的交叉干扰，比传统的多重杂交基因分型（MHG）方法的检测能力提高了10至100倍。此外，SSNB方法在准确区分10种不同的HEV类型和其他常见临床病毒方面表现出100%的特异性。在对70份临床咽拭子样本的分析中，SSNB方法对阳性样本的检测率（50%）略高于逆转录聚合酶链反应（RT-PCR）方法（48.6%）。此外，使用测序方法对经SSNB鉴定为阳性的样本进行分型准确性的进一步评估，结果显示一致性率为100%。该方法兼具高灵敏度和高特异性，同时具备多种类型分析能力且与临床工作流程兼容，使其成为临床环境中一种很有前景的工具。
