Development and Evaluation of a Sensitive PCR-ELISA for Detection of Adenoviruses in Feces

Metzger-Boddien, Christoph; Kehle, Johannes

doi:10.1159/000085098

“…Although false-negative results have been reported [69], these molecular methods have improved sensitivity over traditional methods [62,[70][71][72][73][74]. Any time that adenovirus disease is considered, blood specimens should be sent for quantitative viral load measurements; PCR detection of adenovirus can also be performed on stool, sputum, and biopsy specimens [27,65,68,70,71,[73][74][75]. Quantitative viral load measurements contribute to the diagnosis of infection and act as a surrogate that correlates with clinical response to therapy [67,68].…”

Section: Diagnosis and Monitoringmentioning

confidence: 99%

Emerging Infections: Adenovirus Infections in Transplant Recipients

Ison

¹

2006

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Adenoviruses are increasingly recognized as contributors to morbidity and mortality among stem cell and solid-organ transplant recipients. Clinical presentations range from asymptomatic viremia to respiratory and gastrointestinal disease, hemorrhagic cystitis, and severe disseminated illness. The limited clinical data available support the use of cidofovir for many of these illnesses. Prospective studies are needed to better understand the pathogenesis of and therapeutic options for adenoviral infections in this patient population.

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“…Because the transactivating region of the E1A gene and the N-terminal region of the hexon gene are well conserved between serotypes [60,61], traditional and quantitative real-time PCR techniques have been developed for diagnosis [10,[62][63][64][65][66][67][68]. Although false-negative results have been reported [69], these molecular methods have improved sensitivity over traditional methods [62,[70][71][72][73][74]. Any time that adenovirus disease is considered, blood specimens should be sent for quantitative viral load measurements; PCR detection of adenovirus can also be performed on stool, sputum, and biopsy specimens [27,65,68,70,71,[73][74][75].…”

Section: Diagnosis and Monitoringmentioning

confidence: 99%

“…Although false-negative results have been reported [69], these molecular methods have improved sensitivity over traditional methods [62,[70][71][72][73][74]. Any time that adenovirus disease is considered, blood specimens should be sent for quantitative viral load measurements; PCR detection of adenovirus can also be performed on stool, sputum, and biopsy specimens [27,65,68,70,71,[73][74][75]. Quantitative viral load measurements contribute to the diagnosis of infection and act as a surrogate that correlates with clinical response to therapy [67,68].…”

Section: Diagnosis and Monitoringmentioning

confidence: 99%

Adenoviral Infections in Transplant Recipients

Anderson

¹

,

Ison

²

2014

Emerging Infections 8

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Adenoviruses are increasingly recognized as contributors to morbidity and mortality among stem cell and solid-organ transplant recipients. Clinical presentations range from asymptomatic viremia to respiratory and gastrointestinal disease, hemorrhagic cystitis, and severe disseminated illness. The limited clinical data available support the use of cidofovir for many of these illnesses. Prospective studies are needed to better understand the pathogenesis of and therapeutic options for adenoviral infections in this patient population. VIROLOGY AND EPIDEMIOLOGY Adenoviruses (AdV) are nonenveloped, double-stranded DNA viruses associated with a wide range of clinical syndromes in humans [1]. There are 51 immunologically distinct types of adenoviruses, which are further classified into 1 of 6 (A-F) subgroups on the basis of hemagglutinin properties, DNA homology, oncogenic potential in rodents, and clinical disease (table 1) [2, 3]. Adenoviruses typically cause self-limited respiratory, gastrointestinal, or conjunctival disease throughout the year, without significant seasonal variation. Adenovirus infections are most common among children, people living in close quarters (such as college students and military recruits), and immunocompromised patients. Transmission can occur via inhalation of aerosolized droplets, direct conjunctival inoculation, fecal-oral spread, or exposure to infected tissue or blood [1]. The incubation period depends on the virus serotype and mechanism of transmission and can range from 2 days to 2 weeks [1]. Adenoviruses can establish lifelong asymptomatic infections in lympho-epithelial tissues [4-6]. Among transplant recipients, adenoviral infection has been associated with both de novo infection, particularly in pediatric recipients, and reactivation of latent infection [7, 8]; compelling circumstantial data suggest that most adenovirus infections in adults and subgroup C infections in children arise from reactivation. In both stem cell transplant

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“…14 Alternatively, enzyme-linked immunosorbent assay for colourimetric detection of DIG-labelled PCR amplicons (PCR-ELISA) was developed for the detection of pathogenic microorganisms in the past decades. [15][16][17] PCR-ELISA methods allow the semi-quantitative detection of DNA on a large number of samples simultaneously. 18 In standard PCR-ELISA assays, target specic biotinylated probes immobilized on the streptavidincoated microtiter plate are used to hybridize the DIG-labelled PCR amplicons.…”

Section: Introductionmentioning

confidence: 99%