Improved detection of rotavirus shedding by polymerase chain reaction

Wilde, James A.; Yolken, Robert H.; Eiden, Joseph

doi:10.1016/0140-6736(91)90945-l

Cited by 90 publications

(39 citation statements)

References 17 publications

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“…Although rotavirus has been recovered from patients in some epidemics of NEC, this gastrointestinal virus has not been noted in most patients. Standard methods to detect the rotavirus using enzyme-linked assays may be relatively insensitive, as noted by Wilde et al (8). These investigators were able to demonstrate the presence of rotavirus by reverse transcriptasepolymerase chain reaction when the enzyme immunoassay was negative.…”

mentioning

confidence: 68%

“…In the past, the ability to study viral infections of the gastrointestinal tract has been limited by the unavailability of sensitive methods for viral detection (8). The recent developments of sensitive polymerase chain reaction assays and other new molecular diagnostic tests for the accurate detection of enteric pathogens represent important new tools for the efficient study of enteric pathogens in infants with NEC.…”

Section: Possible Role Of Infectious Agents Inmentioning

confidence: 99%

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Necrotizing Enterocolitis: Research Agenda for a Disease of Unknown Etiology and Pathogenesis

1993

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mentioning

confidence: 68%

Section: Possible Role Of Infectious Agents Inmentioning

confidence: 99%

Necrotizing Enterocolitis: Research Agenda for a Disease of Unknown Etiology and Pathogenesis

1993

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“…However, the collection of large numbers of serum samples from infants, where seasonality in IgM prevalence should be observed, is extremely difficult. These studies should not only be investigated serologically but also through the detection of the viral genome using RT-PCR which could also be genotype-specific [38][39][40]. Significant differences in the distribution of serotypes causing clinical disease in adults versus children have been observed in the UK [40] possibly reflecting different epidemiological transmission processes.…”

Section: Discussionmentioning

confidence: 99%

Serological survey of anti-group A rotavirus IgM in UK adults

Cox

Medley

2003

Epidemiol. Infect.

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SUMMARYRotaviral associated disease of infants in the UK is seasonal and infection in adults not uncommon but the relationship between these has been little explored. Adult sera collected monthly for one year from routine hospital samples were screened for the presence of anti-group A rotavirus immunoglobulin M class antibodies as a marker of recent infection. Anti-rotavirus IgM was seen in all age groups throughout the year with little obvious seasonal variation in the distribution of antibody levels. IgM concentrations and the proportion seropositive above a threshold both increased with age with high concentrations consistently observed in the elderly. Results suggest either high infection rates of rotavirus in adults, irrespective of seasonal disease incidence in infants, IgM persistence or IgM cross-reactivity. These results support recent evidence of differences between infant and adult rotavirus epidemiology and highlight the need for more extensive surveys to investigate age and time related infection and transmission of rotavirus.

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“…Current PCR inhibitor(s) in feces and modified a previously diagnostic techniques, including enzyme-linked im-reported PCR protocol for another member of the fammunosorbent assay (ELISA), polyacrylamide gel elec-ily Reoviridae, 1 resulting in a very sensitive PCR protrophoresis, nucleic acid hybridization, electron mi-tocol for detection of bovine group A rotaviruses in croscopy (EM), and immune electron microscopy, feces. possess limited sensitivity 2,13,15,16,23,[27][28][29] because they generally require 10 6 -10 8 viral particles/ml in fecal Materials and methods samples for detection. Different commercial ELISAs Virus.…”

mentioning

confidence: 99%

“…3 Group A rotaviruses versely affected the sensitivity of PCR. 13,[27][28][29] Several are the major cause of bovine rotaviral diarrhea. 24 treatments have been introduced to eliminate this Groups B-G do not share group antigen with Group problem, including the uses of DNA binding glass, 29 A rotaviruses and have been called pararotaviruses or cellulose fiber (CF-11), 27 and hydroxyapatite (HA).…”

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confidence: 99%

PCR Detection of Group A Bovine Rotaviruses in Feces

et al. 1993

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Abstract.A polymerase chain reaction (PCR) protocol has been developed for identification of bovine group A rotavirus infection in feces. Primers (20mers) complementary to 3' ends of double-stranded RNA genome segment 6 of bovine rotavirus NCDV strain were synthesized and used in PCR. Bovine rotavirus RNA from infected cell culture was employed to optimize the PCR protocol. Rotavirus-negative fecal samples were spiked with known quantities of bovine rotavirus, and the sensitivity of the PCR assay was determined. Fecal samples were extracted with phenol and treated to eliminate unidentified PCR inhibitor(s) in feces, and PCR was performed. PCR products were either visualized on ethidium bromide-stained agarose gels or detected by chemiluminescent hybridization. The sensitivity of the assay was 6 x 10 4 viral particles/ml of feces with ethidium bromide-stained agarose gel visualization or 6 x 10 2 viral particles/ml of feces with chemiluminescent hybridization. The PCR assay was applied to 18 fecal specimens from clinical cases. All 16 clinical samples that were positive for rotavirus by enzyme-linked immunosorbent assay (ELISA) or by ELISA and electron microscopy (EM) were positive by PCR. The 2 samples that were rotavirus negative by ELISA or by ELISA and EM were also negative on PCR analysis.

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Improved detection of rotavirus shedding by polymerase chain reaction

Cited by 90 publications

References 17 publications

Necrotizing Enterocolitis: Research Agenda for a Disease of Unknown Etiology and Pathogenesis

Necrotizing Enterocolitis: Research Agenda for a Disease of Unknown Etiology and Pathogenesis

Serological survey of anti-group A rotavirus IgM in UK adults

PCR Detection of Group A Bovine Rotaviruses in Feces

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