Development of a PCR-and hybridization-based assay (PCR Adenovirus Consensus®) for the detection and the species identification of adenoviruses in respiratory specimens

Vabret, Astrid; Gouarin, S.; Joannes, Martine; Barranger, C.; Petitjean, J.; Corbet, Sandrine; Brouard, J.; Lafay, F; Duhamel, J.F.; Guillois, B.; Freymuth, F.

doi:10.1016/j.jcv.2004.04.003

Cited by 43 publications

(30 citation statements)

References 21 publications

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“…Several in-house methods for adenovirus quantification in different samples have been described (4,10,12,28), and their use has been validated in different clinical situations (7,11,15,16,20,21,24,27 novirus DNA load detection or quantification using the kit assay and the in-house assay described by Heim et al (12), already in use for clinical diagnosis. qPCR assays were performed using the ABI Prism 7000 system, but the kit assay has also been validated for use with the most commonly used real-time PCR systems, including the LightCycler 530 and instruments 560 (Roche), the SmartCycler instruments (Cepheid), the ABI FAM and VIC instruments (Applied Biosystems), and the Rotor-gene Green and Yellow instruments (Corbett Research).…”

Section: Discussionmentioning

confidence: 99%

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Comparison of In-House Real-Time Quantitative PCR to the Adenovirus R-Gene Kit for Determination of Adenovirus Load in Clinical Samples

Jeulin¹,

Salmon

Bordigoni

et al. 2010

J Clin Microbiol

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In the context of hematopoietic stem cell transplantation, adenovirus infections are associated with relevant mortality and morbidity. Detection of adenovirus DNA by quantitative PCR is the "gold standard" for these patients. A total of 150 samples, namely, 78 whole-blood, 22 cerebrospinal fluid, 24 digestive biopsy, and 26 stool samples, from 29 patients, including 24 hematopoietic stem cell transplant recipients, were tested for the detection of adenovirus using an in-house real-time quantitative PCR assay (A. Heim, C. Ebnet, G. Harste, and P. Pring-Akerblom, J. Med. Virol. 70:228-239, 2003) and the commercially available Adenovirus R-Gene kit. Adenovirus DNA was automatically isolated from whole-blood samples (Magna Pure LC system; Roche) or was manually extracted from other specimens (QIAamp; Qiagen) using the appropriate kit. The intra-and interassay reproducibilities and sensitivities were evaluated with cell culture supernatant dilutions. Of the 150 samples tested, 86 were found to be positive and 55 were found to be negative using both techniques. Nine (6%) discordant results were obtained. In most cases, discrepant results concerned samples with low viral loads. Quantitative results for all concordant positive samples were analyzed using the Spearman correlation test. A good correlation between the results of the in-house assay and those of the kit assay was obtained (r ‫؍‬ 0.95; P < 0.001). Regarding the threshold cycle value for internal control spiked samples, none of the 150 samples tested contained a PCR inhibitor. In conclusion, a relevant correlation of results between the in-house assay and the kit assay, as well as the high-quality reproducibility and sensitivity of the kit assay, warranted its use for follow-up of hematopoietic stem cell transplantation recipients.

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Section: Discussionmentioning

confidence: 99%

“…Several in-house PCRs have been described to detect all or some of the 55 adenovirus serotypes (4,10,12,28) and are of clinical interest for adenovirus detection in immunosuppressed patients, but no kit for quantitative detection of adenovirus was commercially available, until recently.…”

mentioning

confidence: 99%

Comparison of In-House Real-Time Quantitative PCR to the Adenovirus R-Gene Kit for Determination of Adenovirus Load in Clinical Samples

Jeulin¹,

Salmon

Bordigoni

et al. 2010

J Clin Microbiol

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“…Standard AdV PCR was performed according to the published method. 8,9 Real-time AdV PCR has been performed since 2005 according to the published method.…”

Section: Design and Methodsmentioning

confidence: 99%

Disseminated adenovirus infections after allogeneic hematopoietic stem cell transplantation: incidence, risk factors and outcome

Robin¹,

Marque-Juillet²,

Scieux³

et al. 2007

Haematologica

119

104

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We analyzed the factors and outcome of patients with disseminated adenovirus infection (dAdV) after allogeneic hematopoeitic stem cell transplantation (HSCT). Thirty patients with dAdV were identified among 620 allogeneic HSCT recipients. Primary diseases were leukemia (n=17), Fanconi anemia (n=12) or others (n=1). Source of stem cells was unrelated in 28 and related in 2 patients. The graft consisted of peripheral blood (n=3), bone marrow (n=12) and unrelated cord-blood (UCB, n=15). Risk factors for dAdV in unrelated HSCT recipients were previous Fanconi disease (p=0.03) and GVHD (p=0.02) in children, and cord blood source of stem cells (p=0.029) and GVHD (0.024) in adults.

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“…In clinical practice, when the role of Ad is suspected in cases of ARI, the diagnosis is currently based on viral isolation techniques, immunofluorescence and, enzymelinked immunosorbent assays (Vabret et al 2004). Polymerase chain reaction (PCR), real-time PCR and immunochromatographic kits have recently become available for this purpose (Fujimoto et al 2004).…”

mentioning

confidence: 99%

Antigenic and genomic characterization of adenovirus associated to respiratory infections in children living in Northeast Brazil

Moura¹,

Mesquita

Portes

et al. 2007

Mem. Inst. Oswaldo Cruz

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Development of a PCR-and hybridization-based assay (PCR Adenovirus Consensus®) for the detection and the species identification of adenoviruses in respiratory specimens

Abstract: The PCR Adenovirus Consensus technique is more efficient than the classical IF or culture techniques for the detection of Ad in respiratory samples. An internal control is included to validate the screening results, and specific probes are used to identify the Ad species.

Cited by 43 publications

References 21 publications

Comparison of In-House Real-Time Quantitative PCR to the Adenovirus R-Gene Kit for Determination of Adenovirus Load in Clinical Samples

Comparison of In-House Real-Time Quantitative PCR to the Adenovirus R-Gene Kit for Determination of Adenovirus Load in Clinical Samples

Disseminated adenovirus infections after allogeneic hematopoietic stem cell transplantation: incidence, risk factors and outcome

Antigenic and genomic characterization of adenovirus associated to respiratory infections in children living in Northeast Brazil

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