Simplified Microneutralization Test for Serotyping Adenovirus Isolates

Malasig, Marietta D.; Goswami, Pulak R.; Crawford-Miksza, Leta; Schnurr, David P.; Gray, Gregory C.

doi:10.1128/jcm.39.8.2984-2986.2001

Cited by 25 publications

(18 citation statements)

References 7 publications

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“…A 20% random selection of adenovirus isolates recovered from sampled recruits with FRI were serotyped using a modified version of the conventional tube neutralization test with flat-bottom 96-well microplates (type-specific rabbit hyperimmune sera to adenoviruses 1-5, 7, and 21, provided by Dr. David Schnurr, Viral and Rickettsial Laboratory, California Department of Health Services, Berkeley, CA) [37]. Beginning in 2002, serotypes were determined through molecular serotyping methods validated against the microneutralization gold standard above [38,39].…”

Section: Laboratory Processingmentioning

confidence: 99%

Vaccine-preventable adenoviral respiratory illness in US military recruits, 1999–2004☆

Russell

Hawksworth

Ryan

et al. 2006

Vaccine

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128

109

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Background and Methods: The high burden of respiratory infections in military populations is well documented throughout history. The primary pathogen responsible for morbidity among US recruits in training was shown to be adenovirus. Highly efficacious oral vaccines were used for 25 years, but vaccine production ceased in 1996, and available stores were depleted by early 1999. Surveillance for acute febrile respiratory illness was performed at eight military recruit training sites throughout the United States from July 1999 through June 2004 to document rates after loss of the vaccines. Laboratory diagnoses complimented the surveillance efforts.

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Section: Laboratory Processingmentioning

confidence: 99%

Vaccine-preventable adenoviral respiratory illness in US military recruits, 1999–2004☆

Russell

Hawksworth

Ryan

et al. 2006

Vaccine

Self Cite

128

109

View full text Add to dashboard Cite

show abstract

“…All positive controls were previously grown in the College of American Pathologists (CAP)-certified NHRC virology laboratory, and all were originally serotyped by the Centers for Disease Control and Prevention (CDC), the American Type Culture Collection (ATCC), or microneutralization (22) in the NHRC laboratory.…”

Section: Controlsmentioning

confidence: 99%

PCR Analysis of Egyptian Respiratory Adenovirus Isolates, Including Identification of Species, Serotypes, and Coinfections

et al. 2005

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Eighty-eight adenovirus (Ad) isolates and associated clinical data were collected from walk-in patients with influenza-like illness in Egypt during routine influenza surveillance from 1999 through 2002. Respiratory Ad distributions are geographically variable, and serotype prevalence has not been previously characterized in this region. Serotype identity is clinically relevant because it predicts vaccine efficacy and correlates strongly with both clinical presentation and epidemiological pattern. Species and serotype identities were determined using several well-validated multiplex PCR protocols culled from the literature and supplemented with a few novel primer sets designed to identify rare types. The isolates included common species B1 serotypes (Ad3 and Ad7), common species C serotypes (Ad1, Ad2, and Ad5), the less common species B2 serotype Ad11, and three isolates of the rare species B1 serotype Ad16. Two isolates that appear to be variant Ad16 were also identified. Fifteen coinfections of multiple adenoviral types, primarily AdB/AdC and Ad3/Ad7 dual infections, were detected. The majority of these were verified using redundant PCR tests targeted at multiple genes. PCR is able to resolve coinfections, in contrast to traditional serum neutralization tests. PCR is also comparatively rapid and requires very little equipment. Application of the method allowed an inclusive determination of the serotypes found in the Egyptian respiratory sample set and demonstrated that coinfections are common and may play a previously unrecognized role in adenovirus pathogenesis, evolution, and epidemiology. In particular, coinfections may influence adenoviral evolution, as interserotypic recombination has been identified as a source of emerging strains.

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“…With WT-Ad, cell lines that support replication are needed, such as Hep2, A549, and 293 cells. The readout is usually either performed microscopically by scoring the Ad-mediated cytopathic effect (CPE) (15), or it is quantifiable by staining for cell viability (3,16). The results from such Ad replication inhibition assays are highly dependent on the timing of readout and usually take from 4 to 8 days.…”

mentioning

confidence: 99%

Quantifying Adenovirus-Neutralizing Antibodies by Luciferase Transgene Detection: Addressing Preexisting Immunity to Vaccine and Gene Therapy Vectors

et al. 2003

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The presence of various levels of anti-adenovirus serotype 5 (Ad5)-neutralizing antibodies in humans is thought to contribute to the inconsistent clinical results obtained so far in diverse gene transfer and vaccination studies and might preclude universal dosing with recombinant Ad5. Prescreening of individuals eligible for Ad5 or alternative serotype treatment and subsequently tailoring the vector dose might aid in ensuring the consistency of clinical parameters. For this purpose, a qualified Ad neutralization assay is required. Here we have tested the different protocols used to date to determine anti-Ad neutralizing activity. Based on simplicity, speed, high throughput, sensitivity, and robustness, we propose a qualified assay in which Ad neutralization is monitored by luciferase reporter gene expression.

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Simplified Microneutralization Test for Serotyping Adenovirus Isolates

Cited by 25 publications

References 7 publications

Vaccine-preventable adenoviral respiratory illness in US military recruits, 1999–2004☆

Vaccine-preventable adenoviral respiratory illness in US military recruits, 1999–2004☆

PCR Analysis of Egyptian Respiratory Adenovirus Isolates, Including Identification of Species, Serotypes, and Coinfections

Quantifying Adenovirus-Neutralizing Antibodies by Luciferase Transgene Detection: Addressing Preexisting Immunity to Vaccine and Gene Therapy Vectors

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