Growth and characterisation of human faecal astrovirus in a continuous cell line

Willcocks, M. M.; Carter, M. J.; Laidler, F. R.; Madeley, C. R.

doi:10.1007/bf01318354

Cited by 111 publications

(67 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Several samples were inoculated and passaged in CaCO2 cells as described by Willcocks and colleagues [15]. Astroviruses were identified by EM and typed by IEM.…”

Section: Cell Culturementioning

confidence: 99%

Identification of astrovirus serotypes from children treated at the Hospitals for Sick Children, London 1981–93

Noel

Cubitt

1994

Epidemiol. Infect.

View full text Add to dashboard Cite

SUMMARYAn enzyme immunoassay (EIA) for astrovirus type 1 together with immune electronmicroscopy (IEM) was used to type a collection of 162 astroviruses obtained from 1981-93 from children with diarrhoea. The EIA was found to be specific for astrovirus type 1. Astrovirus types 2-4 were typed by IEM.Astrovirus type 1 was the prevalent serotype 107/125 (86 %), followed by type 3 (8 %), type 4 (6 %) and type 2 (1 %). Six samples containing astrovirus could not be typed or detected by EIA because they were coated with coproantibodies; 11 others were not identified. Virus particles could no longer be detected in 15/162 (9%) samples following storage for > 2 years.

show abstract

“…Several samples were inoculated and passaged in CaCO2 cells as described by Willcocks and colleagues [15]. Astroviruses were identified by EM and typed by IEM.…”

Section: Cell Culturementioning

confidence: 99%

Identification of astrovirus serotypes from children treated at the Hospitals for Sick Children, London 1981–93

Noel

Cubitt

1994

Epidemiol. Infect.

View full text Add to dashboard Cite

show abstract

“…These studies and subsequent ones based on antigen detection were made possible by use of a monoclonal antibody ELISA able to detect all known human astrovirus serotypes (Herrmann, et al, 1988). This and other improvements in detection techniques, such as better cultivation techniques (Willcocks, et al, 1990) and the application of the reverse transcription polymerase chain reaction (RT-PCR) in numerous studies have been instrumental in defining the epidemiology of this illness worldwide (Cruz, et al, 1992;Dennehy, et al, 2001;Glass, et al, 1996;Glass, 2001;Kotloff, et al, 1992;Maldonado, et al, 1998;Mitchell et al, 1999;Unicomb, et al, 1998;Walter, et al, 2001). RT-PCR is the most sensitive detection method, and in a direct comparison of RT-PCR, ELISA, and EM to monitor an outbreak, it was found that many subclinical infections could only be detected by RT-PCR.…”

Section: Introductionmentioning

confidence: 99%

A DNA oligonucleotide microarray for detecting human astrovirus serotypes

Brown

Gunning

Henry

et al. 2008

Journal of Virological Methods

View full text Add to dashboard Cite

Human astroviruses have been shown in numerous studies to be an important cause of gastroenteritis in young children worldwide. The present communication addresses their characterization by use of oligonucleotide microarray hybridization. The system developed consists of an RT-PCR using primers of low degeneracy capable of detecting all eight serotypes of human astroviruses. RT-PCR products are then hybridized against a microarray consisting of short oligonucleotide probes 17 to 18 nucleotides in length. Cy3-labeled ssDNA targets are generated using a Cy3-labeled primer in the RT-PCR. The non-labeled strand is enzymatically digested, and the labeled target is rescued by column purification. This method of generating labeled target uses equimolar concentrations of the amplifying primers and does not compromise assay sensitivity for initial detection of the virus. Hybridization can be performed without the need for additional amplification. Although the amplicon spans a relatively conserved region of the astrovirus genome, the use of short probes enables type distinction despite such limited diversity. Probes differing by as little as a single nucleotide can be used to distinguish isolates. The microarray developed was capable of distinguishing representatives of the eight known serotypes of human astroviruses.

show abstract

“…We have described the isolation of human astroviruses directly from primary clinical specimens obviating the need for blind-passage adaptation (Willcocks et al, 1990) and have reported the sequence of the genomic 3' terminus . Others have since determined the complete sequence of the type 2 virus and identified the capsid protein gene Monroe et al, 1993).…”

mentioning

confidence: 99%

“…HAst-1 was grown in CaCo-2 cells directly from a diarrhoeal stool (Willcocks et al, 1990). RNA was isolated 18 h after infection (before the appearance of c.p.e.).…”

mentioning

confidence: 99%

The complete sequence of a human astrovirus

Willcocks

Brown

Madeley

et al. 1994

Journal of General Virology

Self Cite

118

100

View full text Add to dashboard Cite

We have determined the complete genomic sequence of human astrovirus serotype 1 isolated in Newcastle upon Tyne. The genome is 6813 nucleotides long and contains three sequential open reading frames (ORFs). The two closest to the 5' end are linked by a ribosomal frameshifting motif and contain sequence motifs indicative of non-structural virus proteins: serine protease and RNA-dependent RNA polymerase. A nuclear addressing sequence is also located here. The 3' ORF encodes the virion structural polypeptides as a polyprotein precursor. This genomic organization resembles that of the plant virus family Luteoviridae.

show abstract

Growth and characterisation of human faecal astrovirus in a continuous cell line

Abstract: We report conditions for the growth of human faecal astrovirus in a continuous colonic carcinoma cell line (CaCo-2). Purified particles contained three polypeptides, one of which (24k) appeared loosely held on the exterior.

Cited by 111 publications

References 19 publications

Identification of astrovirus serotypes from children treated at the Hospitals for Sick Children, London 1981–93

Identification of astrovirus serotypes from children treated at the Hospitals for Sick Children, London 1981–93

A DNA oligonucleotide microarray for detecting human astrovirus serotypes

The complete sequence of a human astrovirus

Contact Info

Product

Resources

About