Feng Wanyu, Chen Yuqing, Han Yanxi, Diao Zhenli, Zhao Zihong, Zhang Yuanfeng, Huang Tao, Ma Yu, Li Ziqiang, Jiang Jian, Li Jing, Li Jinming, Zhang Rui
National Center for Clinical Laboratories, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital/National Center of Gerontology, Beijing, China.
National Center for Clinical Laboratories, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Microbiol Spectr. 2024 Oct 29;12(12):e0164124. doi: 10.1128/spectrum.01641-24.
Respiratory tract infections (RTIs) caused by viruses are prevalent and significant conditions in clinical settings. Accurate and effective detection is of paramount importance in the diagnosis, treatment, and prevention of viral RTIs. With technological advancements, multiplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assays have been developed and extensively adopted for the diagnosis of viral RTIs. Given the potential challenges in the detection performance of multiplex assays, this study evaluated the analytical sensitivity and competitive interference of the six most commonly used multiplex rRT-PCR kits for detection of respiratory viruses in China. The results revealed that the limits of detection were variable across the viruses and kits. Most of the evaluated multiplex kits demonstrated comparable or enhanced analytical sensitivity compared with singleplex kits for clinically significant viruses, including human adenovirus (HAdV)-3, HAdV-7, Omicron BA.5, H1N1pdm09, H3N2, B/Victoria, respiratory syncytial virus subtype A, and respiratory syncytial virus subtype B, whereas multiplex kits showed relatively less analytical sensitivity for human rhinovirus-B72, human metapneumovirus-A2, parainfluenza virus (PIV)-1, and PIV-3. In addition, most multiplex kits successfully identified co-infections when one analyte was present at a low concentration and another analyte was present at a high concentration.
The complexity and severity of viral respiratory tract infections (RTIs) emphasize the pivotal role of precise diagnosis for viral RTIs in guiding effective public health responses and ensuring appropriate medical interventions, given the substantial population at risk. This study highlights the necessity and importance of evaluating the analytical validity of multiplex real-time reverse transcription polymerase chain reaction assays, offering valuable insights into their optimization and application.
由病毒引起的呼吸道感染（RTIs）在临床环境中普遍存在且是重要病症。准确有效的检测对于病毒性RTIs的诊断、治疗和预防至关重要。随着技术进步，多重实时逆转录聚合酶链反应（rRT-PCR）检测方法已被开发并广泛用于病毒性RTIs的诊断。鉴于多重检测在检测性能方面存在潜在挑战，本研究评估了中国六种最常用的多重rRT-PCR试剂盒检测呼吸道病毒的分析灵敏度和竞争性干扰。结果显示，不同病毒和试剂盒的检测限各不相同。与单重试剂盒相比，大多数评估的多重试剂盒对包括人腺病毒（HAdV）-3、HAdV-7、奥密克戎BA.5、甲型H1N1流感（H1N1pdm09）、H3N2、乙型维多利亚（B/Victoria）、呼吸道合胞病毒A亚型和呼吸道合胞病毒B亚型在内的具有临床意义的病毒表现出相当或更高的分析灵敏度，而多重试剂盒对人鼻病毒-B72、人偏肺病毒-A2、副流感病毒（PIV）-1和PIV-3的分析灵敏度相对较低。此外，当一种分析物浓度较低而另一种分析物浓度较高时，大多数多重试剂盒能够成功识别合并感染。
病毒性呼吸道感染（RTIs）的复杂性和严重性凸显了精确诊断病毒性RTIs在指导有效的公共卫生应对措施和确保适当医疗干预方面的关键作用，因为面临风险的人群数量众多。本研究强调了评估多重实时逆转录聚合酶链反应检测方法分析有效性的必要性和重要性，为其优化和应用提供了有价值的见解。
