Enterovirus (EV) infectivity is typically measured as a bulk parameter, yet EV serotypes vary in their susceptibility to natural and engineered stressors. Here we developed an integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) method to simultaneously and specifically quantify the infectious concentrations of eight EV serotypes commonly encountered in sewage (coxsackieviruses A9, B1, B2, B3, B4 and B5, and echoviruses 25 and 30). The method uses two cell lines for virus replication and serotype-specific qPCR primers for quantification. Primers were designed to target multiple environmental strains of a given serotype and displayed high specificity. The ICC-RTqPCR method exhibited a linear calibration range between 50 and 1000 (echoviruses) or 5000 (coxsackieviruses) infectious units per mL. Over this range, measurements were not influenced by the presence of non-target serotypes, and calibration slopes were reproducible for different virus batches and cell ages. The ICC-RTqPCR method was able to accurately quantify the infectious concentration of a virus after inactivation by heat, and the concentration of a virus within a wastewater matrix. This method will be valuable to assess the differing fates of EV serotypes in natural or engineered systems, and to portray the associated changes in EV population composition.

肠道病毒 (EV) 感染性通常作为一个整体参数来衡量，但 EV 血清型对自然和工程压力源的易感性各不相同。在这里，我们开发了一种集成的细胞培养逆转录酶定量 PCR (ICC-RTqPCR) 方法，以同时且特异性地量化污水中常见的八种 EV 血清型（柯萨奇病毒 A9、B1、B2、B3、B4 和 B5，以及埃可病毒 25）的感染浓度和 30）。该方法使用两种细胞系进行病毒复制，使用血清型特异性 qPCR 引物进行定量。引物旨在针对给定血清型的多种环境菌株，并显示出高特异性。ICC-RTqPCR 方法的线性校准范围介于 50 和 1000（回声病毒）或 5000（柯萨奇病毒）感染单位/mL 之间。在这个范围内，测量不受非目标血清型存在的影响，并且校准斜率对于不同的病毒批次和细胞年龄是可重现的。ICC-RTqPCR 方法能够准确量化热灭活后病毒的感染浓度，以及废水基质中病毒的浓度。这种方法对于评估自然或工程系统中 EV 血清型的不同命运，以及描绘 EV 种群组成的相关变化将是有价值的。