Respiratory Adenovirus Quantification with a Droplet Digital Polymerase Chain Reaction (ddPCR) Assay

Abdullah, Omar; Fall, Amary; Forman, Michael; Howard, Craig D.; Klein, Eili Y.; Mostafa, Heba H.

doi:10.1128/spectrum.00269-23

Cited by 5 publications

(7 citation statements)

References 27 publications

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“…Admissions and the need for supplemental oxygen were less likely in patients infected with HAdV-B3 when compared to patients infected with other types, an observation that did not reach statistical significance. The age group and percentage of admissions observed in our study for patients infected with HAdV-non-B3 genotypes were consistent with what we reported for HAdV infections in the time frame of 2020–2022 when HAdV-C predominated ( 8 ). Older age and immunosuppression were the top two variables associated with increased likelihood of admissions with HAdV.…”

Section: Discussionsupporting

confidence: 91%

“…Although higher viral loads in respiratory samples can be associated with acute infections, when comparing HAdV-B3 samples to non-B3 types collected within the first 7 days of symptoms, HAdV-B3 was associated with higher viral loads. Interestingly, in a prior study, we did not detect any significant differences in viral loads between samples from patients infected with different HAdV genotypes ( 8 ). The predominant genotypes in our prior study included HAdV-C1, C2, and C5 ( 8 ).…”

Section: Discussioncontrasting

confidence: 52%

“…Species B and C dominated in a study from Argentina that characterized the circulating types in the period between 2000 and 2018 ( 7 ). Our genomic surveillance of HAdV types from December 2020 to April 2022 showed a predominance of C1 (49.5%) and C2 (34.3%) types and no detection of HAdV-B3 ( 8 ).…”

Section: Introductionmentioning

confidence: 74%

“…Protocol and primer sequences were previously detailed ( 8 ). Samples were extracted via the chemagic Viral RNA/DNA Kit following the manufacturer’s instructions (300 µL extracted volume and 60 µL elution volume).…”

Section: Methodsmentioning

confidence: 99%

“…The hexon gene sequences of the samples screened positive for HAdV were obtained by nested PCR amplification as described previously ( 8 , 11 ). Briefly, for the first PCR , the master mix is composed of dH 2 O (33.75 µL), 10× PCR buffer (5.0 µL), dNTP (4.0 µL at 200 µM), Forward Primer 1 (1.0 µL at 10 µM), Reverse Primer 1 (1.0 µL at 10 µM), and Taq polymerase (Roche Diagnostics) (0.25 µL) for a total of 45 µL per sample.…”

Section: Methodsmentioning

confidence: 99%

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Increased circulation of human adenovirus in 2023: an investigation of the circulating genotypes, upper respiratory viral loads, and hospital admissions in a large academic medical center

Abdullah,

Fall,

Klein

et al. 2024

J Clin Microbiol

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An increase in the circulation of human adenoviruses (HAdV) in 2023 was notable. HAdV genotypes circulating were characterized. Viral loads, clinical presentations, and outcomes were associated with the genotypes. Remnant respiratory samples positive for HAdV after standard-of-care testing at the Johns Hopkins Microbiology laboratory ( N = 270) were collected for genotyping by next-generation sequencing of the hexon gene. HAdV loads in respiratory samples were assessed using droplet digital PCR. The association between predominant genotypes, outcomes, and viral loads was evaluated. Of a total of 249 samples with characterized HAdV genotypes, 179 (71.9%) were genotype B3. HAdV-B3 was associated with a statistically significant increase in viral loads in respiratory samples, specifically in patients 5 years and younger. Patients infected with HAdV-B3 were primarily in the age group 3–5 years in contrast to patients infected with non-B3 genotypes who were younger than 3 years. Strict criteria for defining HAdV-related admission identified a hospitalization rate of 14.8%. Infections with HAdV-B3 were not associated with an increased likelihood of HAdV-related admissions. The circulation of HAdV-B3 in 2023 after at least 2 years of reduced detection likely contributed to the increased number of cases. IMPORTANCE The circulation of human adenoviruses (HAdV) increased in 2023. In this manuscript, we show that HAdV-B3 was predominant in 2023 in a cohort characterized by the Johns Hopkins Hospital System. We also show that HAdV-B3 was associated with an increase in viral loads in respiratory samples and provide a correlation with the clinical presentations and outcomes.

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Section: Discussionsupporting

confidence: 91%

Section: Discussioncontrasting

confidence: 52%

Section: Introductionmentioning

confidence: 74%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Increased circulation of human adenovirus in 2023: an investigation of the circulating genotypes, upper respiratory viral loads, and hospital admissions in a large academic medical center

Abdullah,

Fall,

Klein

et al. 2024

J Clin Microbiol

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Comparison of RT-ddPCR and RT-qPCR platforms for SARS-CoV-2 detection: Implications for future outbreaks of infectious diseases

Ding,

Xu,

Deng

et al. 2024

Environment International

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Adenovirus infections: new insights for the clinical laboratory

Kajon

2024

J Clin Microbiol

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Since their discovery in 1953, research on human adenoviruses (HAdVs) has had diverse foci, resulted in groundbreaking discoveries, such as gene splicing, and generated powerful oncolytic constructs and expression vectors for vaccine development and gene therapy. In contrast, virologists working in this field have made relatively little progress toward the prevention and treatment of the wide spectrum of HAdV-associated diseases. The understanding of species-specific features of viral pathogenesis, or of the mechanisms underlying the establishment of latency and reactivation, is still limited. This group of viruses currently comprises 7 species, 51 serotypes, and 116 unique genotypes. This complexity manifests with a challenging pathophenotypic diversity. Some types are highly virulent, and others do not seem to cause disease in immunocompetent hosts. The assessment of viral load in blood and respiratory specimens has well-acknowledged clinical utility, but the lack of virus typing capabilities easily implementable in clinical laboratories represents a lingering major limitation to the interpretation of positive tests. Some HAdV infections do have severe consequences for both immunocompetent and immunocompromised patients, and the understanding of why this is the case will require more research. Clinical isolates and collections of positive specimens can provide unique resources to investigate the molecular bases of viral virulence and fitness and also help gather information of spatial–temporal patterns of viral circulation in susceptible communities, but they are extremely scarce. Clinical laboratories are underutilized interfaces between patients and academic scientists and have, therefore, a high potential to become valuable collaborators in research moving forward.

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Respiratory Adenovirus Quantification with a Droplet Digital Polymerase Chain Reaction (ddPCR) Assay

Cited by 5 publications

References 27 publications

Increased circulation of human adenovirus in 2023: an investigation of the circulating genotypes, upper respiratory viral loads, and hospital admissions in a large academic medical center

Increased circulation of human adenovirus in 2023: an investigation of the circulating genotypes, upper respiratory viral loads, and hospital admissions in a large academic medical center

Comparison of RT-ddPCR and RT-qPCR platforms for SARS-CoV-2 detection: Implications for future outbreaks of infectious diseases

Adenovirus infections: new insights for the clinical laboratory

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