title: Quantitative reverse transcription PCR to determine the inactivation of Human Rotavirus by chlorine.
abstract: Human rotaviruses (HRVs) are the major cause of acute diarrhea in infants and young children. Here, a real-time reverse transcription polymerase chain reaction assay targeting the rotaviral VP4 gene (VP4-RT-qPCR) was established to evaluate the inactivation of HRV upon chlorine disinfection, based on a previous report that damage to the 1227-2354bp region of the VP4 gene was associated with eliminated HRV infectivity by chlorine. In this study, inactivation of HRV by 0.6mg/L free chlorine was assessed in phosphate buffered saline (PBS; pH 7.2), and tap and river water samples, using both TCID50and RT-qPCR (VP2- and VP4-RT-qPCR) assays, respectively. Among the samples tested, the VP2-RT-qPCR method did not show significant inactivation after chlorine disinfection; however, the reduction in VP4-RT-qPCR signal was correlated with decreased HRV infectivity. Moreover, the higher sensitivity of the VP4-RT-qPCR assay allowed for assessment of chlorine HRV inactivation at longer exposure times compared with the conventional TCID50assay. Collectively, these results indicated that the VP4-RT-qPCR assay is a rapid, sensitive, and reliable tool to detect infectious HRV following chlorine inactivation, and highlights the potential for further development of qPCR/RT-qPCR assays to provide information regarding viral infectivity from drinking water plants.
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xue
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Chenyu
authors.initials: C
authors.lastname: Li
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Tianyu
authors.initials: T
authors.lastname: Zhao
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Qiu
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China.
authors.forename: Min
authors.initials: M
authors.lastname: Jin
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China. Electronic address: wangjingfeng0116@163.com.
authors.forename: Jingfeng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety, No.1 Dali Road, Tianjin, 300050, China. Electronic address: junwen9999@hotmail.com.
authors.forename: Junwen
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics*; D002460:Cell Line; D002713:Chlorine / Q000633:toxicity*; D004203:Disinfection; D008253:Macaca mulatta; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000187:drug effects* / Q000235:genetics; D038301:Virus Inactivation / Q000187:drug effects*
chemical_list: D036022:Capsid Proteins; C060339:VP4 protein, Rotavirus; D002713:Chlorine
keywords: Chlorine; Human rotavirus; Inactivation; Infectivity; RT-qPCR