Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR)

Rigotto, Caroline; Sincero, Thaís Cristine Marques; Simões, C. M. O.; Barardi, Célia Regina Monte

doi:10.1016/j.watres.2004.10.005

Cited by 46 publications

(39 citation statements)

References 24 publications

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“…Cytotoxicity tests were performed to evaluate the potential toxicity caused by seawater in cell lines to be used during disinfection studies. The tests were carried out as previously described (36), with minor modifications. RAW 264.7 and A549 cell monolayers (2 ϫ 10 5 cells/ ml) were propagated in 24-well microplates (Nunc, Rochester, NY) for 24 h before the cytotoxicity assays were performed.…”

Section: Methodsmentioning

confidence: 99%

Comparative Inactivation of Murine Norovirus, Human Adenovirus, and Human JC Polyomavirus by Chlorine in Seawater

Corrêa

Carratalà

Barardi

et al. 2012

Appl Environ Microbiol

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cViruses excreted by humans affect the commercial and recreational use of coastal water. Shellfish produced in contaminated waters have been linked to many episodes and outbreaks of viral gastroenteritis, as well as other food-borne diseases worldwide. The risk can be reduced by appropriate treatment following harvesting and by depuration. The kinetics of inactivation of murine norovirus 1 and human adenovirus 2 in natural and artificial seawater by free available chlorine was studied by quantifying genomic copies (GC) using quantitative PCR and infectious viral particles (PFU). Human JC polyomavirus Mad4 kinetics were evaluated by quantitative PCR. DNase or RNase were used to eliminate free genomes and assess potential viral infectivity when molecular detection was performed. At 30 min of assay, human adenovirus 2 showed 2.6-and 2.7-log 10 GC reductions and a 2.3-and 2.4-log 10 PFU reductions in natural and artificial seawater, respectively, and infectious viral particles were still observed at the end of the assay. When DNase was used prior to the nucleic acid extraction the kinetic of inactivation obtained by quantitative PCR was statistically equivalent to the one observed by infectivity assays. For murine norovirus 1, 2.5, and 3.5-log 10 GC reductions were observed in natural and artificial seawater, respectively, while no viruses remained infectious after 30 min of contact with chlorine. Regarding JC polyomavirus Mad4, 1.5-and 1.1-log 10 GC reductions were observed after 30 min of contact time. No infectivity assays were conducted for this virus. The results obtained provide data that might be applicable to seawater used in shellfish depuration.

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Section: Methodsmentioning

confidence: 99%

Comparative Inactivation of Murine Norovirus, Human Adenovirus, and Human JC Polyomavirus by Chlorine in Seawater

Corrêa

Carratalà

Barardi

et al. 2012

Appl Environ Microbiol

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“…Many PCR primer pairs have been designed and used to target the hexon gene of HAdVs in diverse water sources using conventional, nested, and/or multiplex PCR (2,3,5,6,13,24,32,33,34,41,44,46,53,57). In addition, integrated cell culture PCR (ICC-PCR) techniques have been used to detect infectious HAdV (7,9,21,29,43). However, these methods do not allow direct quantification of HAdVs, impeding their relevance to health risk assessment (10).…”

mentioning

confidence: 99%

Occurrence of Human Adenoviruses at Two Recreational Beaches of the Great Lakes

Xagoraraki¹,

Kuo²,

Wong³

et al. 2007

Appl Environ Microbiol

109

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Human adenoviruses (HAdVs) have been related to several waterborne diseases such as acute gastroenteritis, conjunctivitis, and respiratory illness, and it has been shown that an important human exposure pathway is through recreational waters. However, HAdV occurrence at recreational freshwater beaches has not been previously investigated. In this study, a total of 58 water samples were collected from two recreational beaches on Lake Michigan (

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“…There is evidence that some viral pathogens may be more resistant to environmental conditions and sewage or water treatment processes compared to bacterial organisms (Mena & Gerba 2009). Several reports have previously described the use of molecular methods to evaluate enteric virus contamination in oysters (Rigotto et al 2005, Sincero et al 2006. However, these methods are limited because genome detection by enzymatic amplification does not always accurately predict the contamination risk.…”

Section: Discussionmentioning

confidence: 99%

“…Viruses and cells -The HAV (strain HM 175) was propagated in a continuous line of foetal FRHk-4 cells (Sincero et al 2006) and the HAdV5 was propagated in Hep-2 and A549 cells (Rigotto et al 2005). The cells were cultured in Eagle's minimal essential medium (MEM) (Cultilab, Campinas, SP, Brazil) supplemented with 10% (v/v) foetal bovine serum (FBS) (Gibco-BRL, Life Technologies do Brazil Ltda, SP, Brazil), streptomycin (100 µg/mL), penicillin G (100 U/mL) and amphotericin B (0,025 µg/mL) (Gibco-BRL).…”

Section: Methodsmentioning

confidence: 99%

“…The polymerase chain reaction (PCR) method has become a major tool for virus detection and several studies have shown that PCR can detect fastidious viruses isolated from water samples and molluscs , Ko et al 2005, Rigotto et al 2005, Sincero et al 2006. However, PCR only detects the presence for viral genomes, which does not yield any information about viral infectivity.…”

mentioning

confidence: 99%

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The depuration dynamics of oysters (Crassostrea gigas) artificially contaminated with hepatitis A virus and human adenovirus

Corrêa

Rigotto

Moresco

et al. 2012

Mem. Inst. Oswaldo Cruz

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Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR)

Cited by 46 publications

References 24 publications

Comparative Inactivation of Murine Norovirus, Human Adenovirus, and Human JC Polyomavirus by Chlorine in Seawater

Comparative Inactivation of Murine Norovirus, Human Adenovirus, and Human JC Polyomavirus by Chlorine in Seawater

Occurrence of Human Adenoviruses at Two Recreational Beaches of the Great Lakes

The depuration dynamics of oysters (Crassostrea gigas) artificially contaminated with hepatitis A virus and human adenovirus

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