Detection and sequence-based typing of human adenoviruses using sensitive universal primer sets for the hexon gene

Okada, Morio; Ogawa, Tomoko; Kubonoya, Hiroko; Yoshizumi, Hiroaki; Shinozaki, K.

doi:10.1007/s00705-006-0842-8

Cited by 51 publications

(33 citation statements)

References 23 publications

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“…Nested PCR amplification of a ∼1‐kb partial coding sequence of the adenovirus hexon hypervariable regions HVR1 to HVR‐6, which correlate closely with virus serotype, was carried out using two outer primers (Adhex‐GT3F and Adhex‐GT2R) and two inner primers (Adhex‐GT2F and Adhex‐GT1R), as previously described . Nested PCR products of ∼900 bp were purified using a Wizard SV GEL and PCR Clean‐Up System Kit (Promega Corporation, Madison, WI), and the nucleotide sequences of both DNA strands were then determined by performing direct double‐strand DNA cycle sequencing using the inner primers (Adhex‐GT2F and Adhex‐GT1R) and an ABI PRISM® BigDye® Terminator Cycle Sequencing v3.1 Kit (Applied Biosystems).…”

Section: Methodsmentioning

confidence: 99%

Adenovirus types associated with severe respiratory diseases: A retrospective 4‐year study in Kuwait

Chehadeh

Al-Adwani

John

et al. 2018

Journal of Medical Virology

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Human adenovirus (HAdV) infection can result in a severe respiratory disease. The aim of this study was to identify HAdV types detected in patients hospitalized for severe respiratory illness. The study population consisted of 743 patients with severe respiratory disease admitted to four major hospitals in Kuwait between January 2013 and December 2016. Respiratory specimens were retrospectively screened for 20 respiratory viruses by real-time PCR. The HAdV hexon gene was amplified and directly sequenced, and HAdV types were identified by performing Bayesian phylogenetic analysis. HAdV DNA was detected in 27 (3.6%) patients, with peaks in November and March. Most patients were infants and young children suffering from pneumonia or acute bronchiolitis. The detected HAdV types were C1, C2, C5, B3, and B7. Clusters of HAdV C1, C2, and C5 were observed with high posterior probability. All patients infected with HAdV C5 and 50% of patients infected with HAdV C2 or B7 were admitted to the intensive care unit (ICU). Co-infection with other viruses was detected in 44.4% of patients. The most common co-infecting virus was rhinovirus (HRV). HAdV/HRV co-infection was detected in two children who presumably developed disseminated HAdV infection and died. This is the first report describing the circulation of HAdV types associated with severe outcomes in Kuwait. These findings highlight the need for a national surveillance system to monitor changes in predominant HAdV types and increased numbers of severe respiratory infections.

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Section: Methodsmentioning

confidence: 99%

Adenovirus types associated with severe respiratory diseases: A retrospective 4‐year study in Kuwait

Chehadeh

Al-Adwani

John

et al. 2018

Journal of Medical Virology

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“…The HAdVs belong in the genus of Mastadenovirus of the family of Adenoviridae consisting of more than 60 types [12], grouped into seven species (A to G) based on the serological, biochemical and genetic properties [13, 14]. …”

Section: Introductionmentioning

confidence: 99%

Α molecular epidemiological analysis of adenoviruses from excess conjunctivitis cases

et al. 2017

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BackgroundΤo perform a molecular epidemiological analysis of viral conjunctivitis among excess conjunctivitis cases recorded at the University Hospital of Patras, Greece, for the period March to June 2012.MethodsA structured questionnaire containing demographic and clinical data was developed in order to collect retrospective data on the cases. Eye swab specimens were collected and molecular detection of adenoviruses was performed by nested PCR. Positive results were confirmed by sequencing. To determine the relatedness between the isolated sequences, a phylogenetic analysis was conducted.ResultsThe epidemiological analysis (including retrospective data) included 231 conjunctivitis cases (47.1% male, and 52.8% female). Based on clinical features 205 of the cases were diagnosed of viral origin (46.3% male and 53.7% female), 4 of bacterial origin (50% male and 50% female) while 22 were undefined conjunctivitis. The outbreak excess cases (included 156 cases) affected all age groups regardless gender predilection. For the positive samples indicated that 29 samples (72.5%) were AdV17, and 5 (12.5%) as AdV54.ConclusionsMolecular analysis could define the cause of viral conjunctivitis, while epidemiological data contributed to the assessment of the risk factors and underlined possible preventive measures. This study is one of the very few on viral conjunctivitis in Greece. This outbreak underscores the need for a national surveillance system for acute infectious conjunctivitis outbreaks. The epidemiological as well as molecular investigation on HAdV ocular infections is rather absent in Greece, which has no surveillance system for viral conjunctivitis.

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“…Briefly, the viral DNA of the HAdV-positive samples was extracted from the clinical samples using the QIAamp DNA Mini Kit (Qiagen, Hilden, Germany), following the manufacturer's protocol. Using Ex Taq (TaKaRa Bio Inc., Shiga, Japan), the first PCR was performed with hexon gene-specific primers HX5-3 and HX3-4 (15) and nested PCR was performed with Adhex-GT2F and ADHEX2R (16,17). Electrophoresis was performed in 1.0z agarose gel containing ethidium bromide, and bands were visualized under ultraviolet light.…”

Section: Methodsmentioning

confidence: 99%

Detection and Serotyping of Human Adenoviruses from Patients with Influenza-Like Illness in Mongolia

et al. 2012

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Detection and sequence-based typing of human adenoviruses using sensitive universal primer sets for the hexon gene

Cited by 51 publications

References 23 publications

Adenovirus types associated with severe respiratory diseases: A retrospective 4‐year study in Kuwait

Adenovirus types associated with severe respiratory diseases: A retrospective 4‐year study in Kuwait

Α molecular epidemiological analysis of adenoviruses from excess conjunctivitis cases

Detection and Serotyping of Human Adenoviruses from Patients with Influenza-Like Illness in Mongolia

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