Typing of Human Adenoviruses in Specimens from Immunosuppressed Patients by PCR-Fragment Length Analysis and Real-Time Quantitative PCR

Ebner, K.; Rauch, Margit; Preuner, Sandra; Lion, Thomas

doi:10.1128/jcm.00048-06

Cited by 53 publications

(40 citation statements)

References 33 publications

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“…As seen in other studies (5,9), serotype 2 was the most common isolate. Like Ebner et al (5), we identified serotype 31 without another accompanying serotype (patient 5).…”

Section: Adenovirus (Adv) Infections Are Common In Childhoodsupporting

confidence: 83%

Identification of Adenoviruses in Specimens from High-Risk Pediatric Stem Cell Transplant Recipients and Controls

Zheng

Erdman

et al. 2008

J Clin Microbiol

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Adenovirus infection is an important cause of morbidity and mortality in stem cell transplant recipients. We report species and type-specific analysis from a prospective study of high-risk adenovirus infections following hematopoietic progenitor cell transplantation prior to, during, and after treatment with cidofovir, as well as species analysis of contemporaneously collected samples from control patients. Nine different adenovirus types representing all six recognized species were identified, and mixed infections were commonly found in this group of patients. Adenovirus (ADV) infections are common in childhood.The 51 different currently recognized ADV serotypes are divided into six species (A through F) which have similar genetic and biological properties (3). ADVs commonly cause respiratory, gastrointestinal, and urinary tract disease. In the immunocompromised host, ADV infections can cause severe disease and death. In a recent review, Chakrabarti et al. (3) noted that ADV species B and C account for the majority of clinical isolates in hematopoietic progenitor cell transplantation (HPCT) recipients.In this study, we determined the types of ADVs infecting seven pediatric stem cell transplant recipients prior to, during, and after treatment with cidofovir, as previously described (1), and compared them with 25 consecutive ADV isolates from nontransplant pediatric patients collected contemporaneously to determine the type-specific epidemiology of ADV infections in HPCT recipients and controls. A companion paper (1) examined the epidemiology of ADV infections in HPCT recipients and the effectiveness (if any) of cidofovir against these infections. This study was approved by the IRB of the Children's Memorial Research Center, Children's Memorial Hospital, Chicago, IL.HPCT recipients and specimens. Patients underwent HPCT between 8 September 2003 and 1 December 2005 at our institution. Urine, stool, and throat samples were screened for ADV by viral culture (see below). Plasma was referred to ViraCor (Lee's Summit, MO) for quantitative ADV detection by PCR. Specimens were obtained weekly through the first 100 days (14 weeks) and then monthly until 1 year from the HPCT. A patient was defined as having a high-risk ADV infection (HRAI) if they had (i) a positive blood ADV PCR with or without symptoms, (ii) two or more positive ADV cultures from separate peripheral sites with or without symptoms, or (iii) one positive peripheral culture for ADV plus compatible clinical evidence of ADV (1). As previously described, 7 of the 38 children developed HRAI and received cidofovir treatment (1). Two additional asymptomatic patients had ADV identified transiently from a single body site only without clinical symptoms and were not further studied.Specimens from other patients. Twenty-five consecutive pediatric specimens from nontransplanted patients submitted to our virology laboratory during routine clinical care that grew ADV also had ADV species identification performed. These specimens included those from the respiratory trac...

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“…As seen in other studies (5,9), serotype 2 was the most common isolate. Like Ebner et al (5), we identified serotype 31 without another accompanying serotype (patient 5).…”

Section: Adenovirus (Adv) Infections Are Common In Childhoodsupporting

confidence: 83%

Identification of Adenoviruses in Specimens from High-Risk Pediatric Stem Cell Transplant Recipients and Controls

Zheng

Erdman

et al. 2008

J Clin Microbiol

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“…These results suggested that the PCR/RFLP method described in this paper could be a useful screening approach for AdV detection and at the same time for AdV species typing, using a single restriction enzyme, HaeIII, in contrast to others RFLP strategies which have been used to identify adenoviruses serotypes or species by using several restriction enzymes (Li et al 1999, Elnifro et al 2000, Allard et al 2001, Ebner et al 2006. However, to evaluate the capability of the PCR/RFLP described in this paper to detect the different AdV species is necessary to test more AdV reference strains and isolates, since sequencing of the AdV hexon gene has revealed a divergence at protein level of 0.5 to 25.4% among the 51 serotypes (Ebner et al 2005a), and this variability may explain the unexpected RFLP found with the AdV serotype 41 (species F).…”

Section: Discussionmentioning

confidence: 85%

Adenoviruses C in non-hospitalized Mexican children older than five years of age with acute respiratory infection

Rosete

Manjarrez

Barrón

2008

Mem. Inst. Oswaldo Cruz

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“…Immunotyping methods provide definitive identification of serotype but are excessively time-consuming, labor-intensive, and restricted to a few laboratories that possess reference immune reagents. A variety of molecular methods have been advanced to augment immunotyping, including genome restriction analysis (20), PCR-coupled microarrays (21,22), PCRfragment length analysis (23), electrospray ionization mass spectrometry (24), and more commonly, partial sequencing of specific target genes (25,26). Although an advance over immunotyping methods, these methods require specialized technical skills and utilize technologies that may be of limited availability outside the research laboratory setting.…”

mentioning

confidence: 99%

Quantitative Real-Time PCR Assay Panel for Detection and Type-Specific Identification of Epidemic Respiratory Human Adenoviruses

Trujillo-Lopez²,

Lott³

et al. 2013

J Clin Microbiol

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bOutbreaks of human adenovirus (HAdV) acute respiratory illness (ARI) have been well documented among civilians and unvaccinated military recruits. Among the 7 recognized HAdV species (A to G), species B (particularly serotypes 3, 7, 11, 14, and 21) and E (serotype 4) have more often been associated with epidemic ARI. Rapid detection and type-specific identification of these viruses would enhance outbreak response and help guide prevention and control measures. To this end, we developed type-specific real-time quantitative PCR (qPCR) assays for HAdV types 3, 4, 7, 11, 14, 16, and 21 targeting the HAdV hexon gene. All typespecific qPCR assays reproducibly detected as few as 10 copies/reaction of quantified hexon recombinant plasmids with a linear dynamic range of 8 log units (10 1 to 10 8 copies); in contrast, a generic qPCR assay that detects all HAdV types run concurrently detected between 10 and 100 copies/reaction, depending on the virus type. No nonspecific amplifications were observed with concentrated nucleic acid from 51 HAdV prototype strains or other common respiratory pathogens. All members of a panel of 137 previously typed HAdV field isolates and positive clinical specimens were correctly characterized by the type-specific qPCR assays; two different HAdV types were detected in three of the clinical specimens and confirmed by amplicon sequencing. The qPCR assays permit sensitive, specific, and quantitative detection and identification of seven clinically important respiratory HAdVs and should provide a convenient adjunct to classical typing methods for a rapid response to HAdV outbreaks.

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Typing of Human Adenoviruses in Specimens from Immunosuppressed Patients by PCR-Fragment Length Analysis and Real-Time Quantitative PCR

Cited by 53 publications

References 33 publications

Identification of Adenoviruses in Specimens from High-Risk Pediatric Stem Cell Transplant Recipients and Controls

Identification of Adenoviruses in Specimens from High-Risk Pediatric Stem Cell Transplant Recipients and Controls

Adenoviruses C in non-hospitalized Mexican children older than five years of age with acute respiratory infection

Quantitative Real-Time PCR Assay Panel for Detection and Type-Specific Identification of Epidemic Respiratory Human Adenoviruses

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