Diarrheal diseases account for more than 50% of foodborne diseases worldwide, the majority of which occur in infants and young children. The traditional bacterial detection method is complex and time-consuming; therefore, it is necessary to establish a rapid and convenient detection method that can detect multiple pathogens simultaneously. In this study, we developed a set of five multiplex real-time SYBR Green I PCR assays to simultaneously detect 15 common enteric pathogens based on the Homo-Tag Assisted Non-Dimer system. These assays effectively reduced primer-dimer formation and improved the stability, uniformity, and amplification efficiency of multiplex PCR. The detection limit of the multiplex SYBR Green I PCR system was approximately 10⁴–10⁶ CFU/mL for stool specimens. Furthermore, we vitrified heat-unstable components on the cap of a reaction tube, showing that Taq DNA polymerase, dNTPs, primers, and SYBR Green I remained stable at 25 °C. In summary, we developed multiplex SYBR Green I PCR assays that can simultaneously detect 15 enteric pathogens. This method is comprehensive, rapid, inexpensive, accurate, and simple and displays high specificity.

腹泻病占全世界食源性疾病的50％以上，其中大多数发生在婴幼儿中。传统的细菌检测方法复杂且耗时；因此，有必要建立一种快速便捷的检测方法，以同时检测多种病原体。在这项研究中，我们开发了一套五组多重实时SYBR Green I PCR检测试剂盒，可基于Homo-Tag Assisted Non-Dimer系统同时检测15种常见的肠道病原体。这些测定有效减少了引物二聚体的形成，并提高了多重PCR的稳定性，均匀性和扩增效率。多重SYBR Green I PCR系统的检出限约为10 ⁴ –10 ⁶ 大便标本的CFU / mL。此外，我们将反应管帽上的热不稳定成分玻璃化，显示Taq DNA聚合酶，dNTPs，引物和SYBR Green I在25°C时保持稳定。总之，我们开发了可同时检测15种肠道病原体的SYBR Green I PCR多重检测方法。该方法是全面，快速，廉价，准确，简单的方法，显示出很高的特异性。