Development of a new cell culture-based method and optimized protocol for the detection of enteric viruses

Lee, Jae Ho; Lee, Gyu-Cheol; Kim, Jong Ik; Yi, Hyun; Lee, Chan Hee

doi:10.1016/j.jviromet.2013.03.019

Cited by 11 publications

(6 citation statements)

References 36 publications

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“…Concentrated water samples reserved for assays of infectivity were thawed and maintained at 4 • C, until inoculation into Vero E6 cultures (Vero C 1008, ATCC CRL-1586) [46,47]. Cells were grown at 37 • C in 25 cm 2 flasks (T25) in the following culture medium (culture medium, FBS10): CO 2 -Independent Medium (Gibco, Thermo Fisher Scientific; Waltham, MA, USA) with additional 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific; Waltham, MA, USA), 2 mM GlutaMAX (Gibco, Thermo Fisher Scientific; Waltham, MA, USA), and 0.5 mg/mL gentamicin (Gibco, Thermo Fisher Scientific; Waltham, MA, USA).…”

Section: Infectivity Assaysmentioning

confidence: 99%

Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

et al. 2020

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Hepatitis E virus (HEV) is a non-enveloped single-stranded positive-sense RNA virus, belonging to the Hepeviridae family, resistant to environmental conditions, and transmitted by the consumption of contaminated water. This virus is responsible for both sporadic and epidemic outbreaks, leading to thousands of infections per year in several countries, and is thus considered an emerging disease in Europe and Asia. This study refers to a survey in Portugal during 2019, targeting the detection and eventual quantification of enteric viruses in samples from surface and drinking water. Samples positive for HEV RNA were recurrently found by reverse transcription quantitative PCR (RT-qPCR), in both types of matrix. The infectivity of these samples was evaluated in cultured Vero E6 cells and RNA from putative viruses produced in cultures evidencing cytopathic effects and was subjected to RT-qPCR targeting HEV genomic RNA. Our results evidenced the existence of samples positive either for HEV RNA (77.8% in surface water and 66.7% in drinking water) or for infectious HEV (23.0% in surface water and 27.7% in drinking water). These results highlight the need for effective virological control of water for human consumption and activities.

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Section: Infectivity Assaysmentioning

confidence: 99%

Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

et al. 2020

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“…HAV (ATCC- VR-2266) was grown in MRC-145 cells [ 72 ]. Coxsackie Virus B5 (EV71; ATCC VR-185) was grown in Vero cells [ 73 ]. MUV virus (ATCC VR-1438) was grown in HEp-2 cells [ 74 ].…”

Section: Methodsmentioning

confidence: 99%

A universal mammalian vaccine cell line substrate

et al. 2017

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Using genome-wide small interfering RNA (siRNA) screens for poliovirus, influenza A virus and rotavirus, we validated the top 6 gene hits PV, RV or IAV to search for host genes that when knocked-down (KD) enhanced virus permissiveness and replication over wild type Vero cells or HEp-2 cells. The enhanced virus replication was tested for 12 viruses and ranged from 2-fold to >1000-fold. There were variations in virus-specific replication (strain differences) across the cell lines examined. Some host genes (CNTD2, COQ9, GCGR, NDUFA9, NEU2, PYCR1, SEC16G, SVOPL, ZFYVE9, and ZNF205) showed that KD resulted in enhanced virus replication. These findings advance platform-enabling vaccine technology, the creation of diagnostic cells substrates, and are informative about the host mechanisms that affect virus replication in mammalian cells.

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“…Several methods for detecting RV and AdV are currently available: 1) viruses were cultured in cells, and immunological or molecular biological methods are used to identify the viruses [ 9 ]; 2) immunological methods are used directly to detect the presence of antigens or IgM and IgG antibodies in the specimens [ 10 – 12 ]; and 3) molecular biological methods are used directly to detect antigens [ 10 , 13 ]. Methods 1 and 3 are limited with respect to technology and equipment, especially in primary medical institutions and field tests.…”

Section: Introductionmentioning

confidence: 99%

Comparison of two different combined test strips with fluorescent microspheres or colored microspheres as tracers for rotavirus and adenovirus detection

Jiang

Shi

Lin

et al. 2018

Virol J

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BackgroundRotavirus (RV) and enteric adenovirus (AdV) mainly cause infantile infectious gastroenteritis. Several separate test methods for the detection of RV or AdV are currently available, but few tests are able to simultaneously detect both RV and AdV viruses, especially in primary medical institutions.MethodsThe present study was mainly designed to compare the performance of two combined test strips for the detection of RV and AdV: a rotavirus–adenovirus strip with fluorescent microspheres for tracers (FMT); and the CerTest rotavirus–adenovirus blister strip with colored microspheres for tracers (CMT). To test the strips cultures of RV, AdV and from other enteric pathogens were used, in addition to 350 stool specimens from 45 symptomatic patients with gastrointestinal infections.ResultsDetection thresholds for RV and AdV cultures using serial dilutions showed that the sensitivity of FMT was significantly higher than that of CMT (both P < 0.05). Specificity evaluation demonstrated that with culture mixtures of Coxsackie (A16), ECHO (type30), and entero- (EV71) viruses there was no detection of cross reaction using the two test strips, i.e., all the results were negative. With regard to the detection of RV in 350 clinical specimens, the total coincidence rate was 92.9%, the positive coincidence rate was 98.2%, and the negative coincidence rate was 90.8%. With regard to AdV detection, the total coincidence rate was 95.4%, the positive coincidence rate was 95.2%, and the negative coincidence rate was 95.5%.ConclusionsFMT performed better than CMT with regard to the combined detection of RV and AdV.

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Development of a new cell culture-based method and optimized protocol for the detection of enteric viruses

Cited by 11 publications

References 36 publications

Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

A universal mammalian vaccine cell line substrate

Comparison of two different combined test strips with fluorescent microspheres or colored microspheres as tracers for rotavirus and adenovirus detection

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