An integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) method to simultaneously quantify the infectious concentrations of eight environmentally relevant enterovirus serotypes
نویسندگان
چکیده
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 simultaneously specifically quantify the infectious concentrations of eight commonly encountered sewage (coxsackieviruses A9, B1, B2, B3, B4 B5, echoviruses 25 30). The uses two lines for virus replication serotype-specific qPCR primers quantification. Primers were designed target multiple environmental strains given serotype displayed high specificity. ICC-RTqPCR exhibited linear calibration range between 50 1000 (echoviruses) or 5000 (coxsackieviruses) units per mL. Over this range, measurements not influenced by presence non-target serotypes, slopes reproducible different batches ages. was able accurately concentration after inactivation heat, within wastewater matrix. This will be valuable assess differing fates systems, portray associated changes population composition.
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ژورنال
عنوان ژورنال: Journal of Virological Methods
سال: 2021
ISSN: ['1879-0984', '0166-0934']
DOI: https://doi.org/10.1016/j.jviromet.2021.114225