Quantitative Real-Time PCR Assays for Detection of Human Adenoviruses and Identification of Serotypes 40 and 41

Jothikumar, Narayanan; Cromeans, Theresa L.; Hill, Vincent R.; Lu, Xiaoyan; Sobsey, Mark D.; Erdman, Dean D.

doi:10.1128/aem.71.6.3131-3136.2005

Cited by 236 publications

(160 citation statements)

References 29 publications

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“…The sequences and sources of the primers and probes used in this present study are shown in Table 1 . [28][29][30][31][32] Each assay was validated using positive controls and negative controls consisting of non-target NA and DEPCtreated water.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

Gibson¹,

Opryszko

Schissler

et al. 2011

The American Society of Tropical Medicine and Hygiene

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Abstract. An estimated 884 million people worldwide do not have access to an improved drinking water source, and the microbial quality of these sources is often unknown. In this study, a combined tangential flow, hollow fiber ultrafiltration (UF), and real-time PCR method was applied to large volume (100 L) groundwater ( N = 4), surface water ( N = 9), and finished (i.e., receiving treatment) drinking water ( N = 6) samples for the evaluation of human enteric viruses and bacterial indicators. Human enteric viruses including norovirus GI and GII, adenovirus, and polyomavirus were detected in five different samples including one groundwater, three surface water, and one drinking water sample. Total coliforms and Escherichia coli assessed for each sample before and after UF revealed a lack of correlation between bacterial indicators and the presence of human enteric viruses.

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Section: Methodsmentioning

confidence: 99%

Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

Gibson¹,

Opryszko

Schissler

et al. 2011

The American Society of Tropical Medicine and Hygiene

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“…Based on previous studies, the standard cur ve was generated as follows [7][8][9] : Standard DNA from S. enteritidis was used to establish a standard curve. The standard DNA contained amplified target DNA in different quantities which was included in each LightCycler run.…”

Section: Fq-pcr Standard Curvementioning

confidence: 99%

Gastrointestinal tract distribution of Salmonella enteritidis in orally infected mice with a species-specific fluorescent quantitative polymerase chain reaction

Deng

Cheng²,

Wang³

et al. 2007

WJG

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cecum, without causing apparent symptoms. By 5 d post inoculation, the cecum had significantly higher numbers of S. enteritidis than any of the other areas (P < 0.01), and this appeared to reflect its function as a repository for S. enteritidis. CONCLUSION:The results provided significant data for clarifying the pathogenic mechanism of S. enteritidis in the gastrointestinal tract, and showed that the jejunum, ileum and cecum are the primary sites of invasion in normal mice after oral infection. This study will help to further understanding of the mechanisms of action of S. INTRODUCTIONSalmonella is an enteric pathogen that colonizes the intestinal tract of a variety of animals, especially humans and poultry, and accounts for millions of cases of gastroenteritis and food-borne illness each year [1,2] . Salmonella enteritidis (S. enteritidis) can be transmitted to humans through the food production chain, and undercooked or raw eggs and poultry meat are a particularly high risk for humans [3] . In the last few decades, S. enteritidis has emerged as a major cause of food-borne illness worldwide. As a result of the increased prevalence of S. enteritidis and its complex life cycle, identifying the regular distribution pattern of S. enteritidis in the gastrointestinal tract will help to understand its mechanism of action.Previous studies have shown that orally introduced S. enteritidis has a rapid transit time through the intestine, and a small proportion of the inoculum establishes itself within the walls of the small intestine and cecum several enteritidis. We used this assay to detect genomic DNA of S. enteritidis in the gastrointestinal tract, including duodenum, jejunum, ileum, cecum, colon, rectum, esophagus and stomach, from mice after oral infection. RESULTS

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“…Given the localization of 55R E1A in the nucleus during infection, we sought to assess whether, akin to the largest E1A isoforms, 55R E1A could activate viral gene expression. To accomplish this, we performed a qRT-PCR assay on contact-inhibited human diploid IMR-90 fibroblast cells infected with dl309, JM17-55R, dl521, or dl312, using primers that recognize transcripts expressed from selected viral promoters (1,13,18). The E1A products expressed by each of these viruses are listed in Fig.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the 55-Residue Protein Encoded by the 9S E1A mRNA of Species C Adenovirus

Miller

Pelka

Fonseca

et al. 2012

J Virol

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Early region 1A (E1A) of human adenovirus (HAdV) has been the focus of over 30 years of investigation and is required for the oncogenic capacity of HAdV in rodents. Alternative splicing of the E1A transcript generates mRNAs encoding multiple E1A proteins. The 55-residue (55R) E1A protein, which is encoded by the 9S mRNA, is particularly interesting due to the unique properties it displays relative to all other E1A isoforms. 55R E1A does not contain any of the conserved regions (CRs) present in the other E1A isoforms. The C-terminal region of the 55R E1A protein contains a unique sequence compared to all other E1A isoforms, which results from a frameshift generated by alternative splicing. The 55R E1A protein is thought to be produced preferentially at the late stages of infection. Here we report the first study to directly investigate the function of the species C HAdV 55R E1A protein during infection. Polyclonal rabbit antibodies (Abs) have been generated that are capable of immunoprecipitating HAdV-2 55R E1A. These Abs can also detect HAdV-2 55R E1A by immunoblotting and indirect immunofluorescence assay. These studies indicate that 55R E1A is expressed late and is localized to the cytoplasm and to the nucleus. 55R E1A was able to activate the expression of viral genes during infection and could also promote productive replication of species C HAdV. 55R E1A was also found to interact with the S8 component of the proteasome, and knockdown of S8 was detrimental to viral replication dependent on 55R E1A.

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Quantitative Real-Time PCR Assays for Detection of Human Adenoviruses and Identification of Serotypes 40 and 41

Cited by 236 publications

References 29 publications

Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

Gastrointestinal tract distribution of Salmonella enteritidis in orally infected mice with a species-specific fluorescent quantitative polymerase chain reaction

Characterization of the 55-Residue Protein Encoded by the 9S E1A mRNA of Species C Adenovirus

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