An integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) method to simultaneously quantify the infectious concentrations of eight environmentally relevant enterovirus serotypes

Larivé, Odile; Brandani, Jade; Dubey, Manupriyam; Kohn, Tamar

doi:10.1016/j.jviromet.2021.114225

Cited by 6 publications

(11 citation statements)

References 37 publications

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“…Effect of Host Cells on the Inactivation Kinetics of Other Genotypes of the Enterovirus Genus. Additional enterovirus genotypes (CVA9, CVB1, E7, E9, and E13), which were reported to be prevalent in European sewage, 60 were tested for their ability to infect the three host cells. We found that these viruses can propagate not only in RD and A549 cells but also in BGMK cells, which lack CD55 and β2M expression (Table S2).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Observed Kinetics of Enterovirus Inactivation by Free Chlorine Are Host Cell-Dependent

Torii

David

Larivé

et al. 2023

Environ. Sci. Technol.

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Virucidal efficacies of disinfectants are typically assessed by infectivity assay utilizing a single type of host cell. Enteroviruses infect multiple host cells via various entry routes, and each entry route may be impaired differently by a given disinfectant. Yet, it is unknown how the choice of host cells affects the observed inactivation kinetics. Here, we evaluated the inactivation kinetics of echovirus 11 (E11) by free chlorine, ultraviolet (UV) irradiation, and heat, using three different host cells (BGMK, RD, and A549). Inactivation rates were independent of the host cell for treatment of E11 by UV or heat. Conversely, E11 inactivation by free chlorine occurred 2-fold faster when enumerated on BGMK cells compared with RD and A549 cells. Host cell-dependent inactivation kinetics by free chlorine were also observed for echovirus 7, 9, and 13, and coxsackievirus A9. E11 inactivation by free chlorine was partly caused by a loss in host cell attachment, which was most pronounced for BGMK cells. BGMK cells lack the attachment receptor CD55 and a key subunit of the uncoating receptor β2M, which may contribute to the differential inactivation kinetics for this cell type. Consequently, inactivation kinetics of enteroviruses should be assessed using host cells with different receptor profiles.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Observed Kinetics of Enterovirus Inactivation by Free Chlorine Are Host Cell-Dependent

Torii

David

Larivé

et al. 2023

Environ. Sci. Technol.

Self Cite

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show abstract

“…A panel of enteroviruses (CVA9, CVB1, E7, E9, and E13), which were reported to be prevalent in European sewage 50 , was tested for their ability to infect the three host cells (Table S1). The results suggested that these viruses can propagate in BGMK cells as well as RD and A549 cells, despite the lack of CD55 and β2M.…”

Section: Resultsmentioning

confidence: 99%

Observed Kinetics of Enterovirus Inactivation by Free Chlorine Is Host Cell-Dependent

Torii

David

Larivé

et al. 2022

Preprint

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The virucidal efficacy of disinfectants is typically assessed by infectivity assay utilizing a single type of host cell. Enteroviruses infect multiple host cells via different entry routes, and each entry route may be impaired to a varying extent by a given disinfectant. Yet, it is not known how the choice of host cells for titration affects the observed inactivation kinetics. Here, we evaluated the inactivation kinetics of echovirus 11 (E11) by free chlorine, ultraviolet (UV) irradiation, and heat, using three different host cells (BGMK, RD, and A549). E11 inactivation by free chlorine occurred at a two-fold greater rate when enumerated on BGMK cells compared to RD and A549 cells. Conversely, a comparable inactivation rate was observed for UV and heat independent of the host cell used. Host cell-dependent inactivation kinetics by free chlorine were also observed for echovirus 7, 9 and 13, and coxsackievirus A9, confirming that this phenomenon is not serotype-specific. Inactivation of E11 was partly caused by a loss in host cell attachment, which was most pronounced for BGMK cells, and which may be promoted by a lack of CD55 attachment receptors on this cell type. Additionally, BGMK cells lack a key subunit of the uncoating receptor, β2M, which may further contribute to the differential inactivation kinetic for this cell type. Consequently, inactivation kinetics of enteroviruses should be assessed using host cells with different receptor profiles. This will yield a more complete understanding of the inactivating power of disinfectants targeting the viral attachment and/or uncoating.

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“…The integrated cell culture–reverse transcription–quantitative polymerase chain reaction (ICC-RT-qPCR) assay which combines cell culture with targeted viral genome detection by qPCR was employed to determine the loss of MHV-A59 infectivity during disinfection in a timely and specific manner (Text S6). Synthetic cDNA oligos (Integrated DNA Technologies, Inc.) were prepared in a series of 10-fold dilutions (10 3 –10 12 gene copies mL –1 ) to generate the standard curve of absolute gene copy numbers versus cycle threshold values. In our RT-qPCR protocol, 5 μL of the RNA template was added to 5 μL of TaqMan fast virus one-step master mix (Fisher Scientific, Applied Biosystems 4444434), 2 μL of forward primer (10 μM), 2 μL of reverse primer (10 μM), 1.25 μL of probe (10 μM), and 4.75 μL of nuclease-free water, following cycling conditions of 52 °C for 10 min (reverse transcription), 95 °C for 20 s (reverse transcription inactivation and denaturation initiation), and 45 cycles of 95 °C for 15 s and 60 °C for 1 min.…”

Section: Methodsmentioning

confidence: 99%

Fe–Fe Double-Atom Catalysts for Murine Coronavirus Disinfection: Nonradical Activation of Peroxides and Mechanisms of Virus Inactivation

Zhu

Zhang

et al. 2023

Environ. Sci. Technol.

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Peroxides find broad applications for disinfecting environmental pathogens particularly in the COVID-19 pandemic; however, the extensive use of chemical disinfectants can threaten human health and ecosystems. To achieve robust and sustainable disinfection with minimal adverse impacts, we developed Fe single-atom and Fe−Fe double-atom catalysts for activating peroxymonosulfate (PMS). The Fe−Fe double-atom catalyst supported on sulfur-doped graphitic carbon nitride outperformed other catalysts for oxidation, and it activated PMS likely through a nonradical route of catalyst-mediated electron transfer. This Fe− Fe double-atom catalyst enhanced PMS disinfection kinetics for inactivating murine coronaviruses (i.e., murine hepatitis virus strain A59 (MHV-A59)) by 2.17−4.60 times when compared to PMS treatment alone in diverse environmental media including simulated saliva and freshwater. The molecular-level mechanism of MHV-A59 inactivation was also elucidated. Fe−Fe double-atom catalysis promoted the damage of not only viral proteins and genomes but also internalization, a key step of virus lifecycle in host cells, for enhancing the potency of PMS disinfection. For the first time, our study advances double-atom catalysis for environmental pathogen control and provides fundamental insights of murine coronavirus disinfection. Our work paves a new avenue of leveraging advanced materials for improving disinfection, sanitation, and hygiene practices and protecting public health.

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An integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) method to simultaneously quantify the infectious concentrations of eight environmentally relevant enterovirus serotypes

Cited by 6 publications

References 37 publications

Observed Kinetics of Enterovirus Inactivation by Free Chlorine Are Host Cell-Dependent

Observed Kinetics of Enterovirus Inactivation by Free Chlorine Are Host Cell-Dependent

Observed Kinetics of Enterovirus Inactivation by Free Chlorine Is Host Cell-Dependent

Fe–Fe Double-Atom Catalysts for Murine Coronavirus Disinfection: Nonradical Activation of Peroxides and Mechanisms of Virus Inactivation

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