Digital droplet PCR for the detection and quantification of circulating bovine Deltapapillomavirus

Falco, Francesca De; Corrado, Federica; Cutarelli, Anna; Leonardi, Leonardo; Roperto, Sante

doi:10.22541/au.158879220.04096886

Cited by 2 publications

(3 citation statements)

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“…We detected and sequenced OaPVs in the urinary bladder of healthy cattle (personal observations), which supports the hypothesis that OaPVs may contribute to the composition of the normal bladder microbiota of cattle. BPV and HPV DNA have also been reported in the blood of healthy cattle (De Falco, Corrado, et al., 2021) and asymptomatic blood donors (Vergara et al., 2019). It has been suggested that there is an actual likelihood that HPVs could reach epithelial target sites in the blood, which might explain how and why HPVs are associated with tumours of several organs (Cladel et al., 2019; Conceição Gomes Nascimento et al., 2021; Syrjänen & Syrjänen, 2021; Vergara et al., 2019).…”

Section: Discussionmentioning

confidence: 94%

“…Each sample was analysed in duplicate. According to previous studies on papillomavirus detection and quantification using ddPCR (De Falco et al., 2021a; De Falco et al., 2021b; Jeannot et al., 2016, 2021), blood samples were considered OaPV‐positive in the presence of at least three positive droplets at the same amplitude as positive controls. A sample was considered OaPV‐negative when fewer than three droplets or no droplets containing OaPV amplicons were observed.…”

Section: Methodsmentioning

confidence: 99%

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Evidence of a novel cross‐species transmission by ovine papillomaviruses

Falco

Cutarelli

Cuccaro

et al. 2022

Transbounding Emerging Dis

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Ovine papillomavirus (OaPV) comprises four genotypes; OaPV1, OaPV2 and OaPV4 are fibropapillomaviruses within the genus Deltapapillomavirus, whereas OaPV3 is an epitheliotropic virus that belongs to the genus Dyokappapapillomavirus. To date, all of them have been known to infect sheep only. OaPV1, OaPV2 and OaPV4 have been associated with ovine cutaneous and mucosal fibropapillomas, whereas OaPV3 is a key factor in the squamous cell carcinoma pathway of the sheep skin. Whole blood samples obtained from 128 cattle at public slaughterhouses were investigated using droplet digital polymerase chain reaction (ddPCR). ddPCR is a new‐generation PCR technique that enables an accurate and absolute quantification of target molecules with high sensitivity and specificity. All OaPVs were detected by identification and quantification of nucleic acids using specific fluorescent probes. Of 128 blood samples, 100 (∼78%) showed OaPV infections. Further, 42, 35 and 23 blood samples showed single, double and triple OaPV infections, respectively. OaPV1 was responsible for 22 single infections, OaPV2 caused 16 single infections and OaPV3 and OaPV4 caused two single infections each. OaPV1 and OaPV2 were the most frequent ovine viruses in dual and triple infections. In many blood samples, both ovine deltapapillomavirus and dyokappapapillomavirus were found to be transcriptionally active, as shown by the detection and quantification of E5 oncogene transcripts for OaPV1, L1 transcripts for OaPV2, E6 and E7 transcripts for OaPV3 and E6 for OaPV4. OaPVs were found in the blood samples from cattle that shared grasslands rich in bracken ferns known to contain immunosuppressant substances. Furthermore, OaPVs were also found in cattle from intensive livestock farming without any contact with sheep. Because OaPV DNA was detected in both grass hay and corn silage, it is conceivable that these feed may be the viral sources.

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Section: Discussionmentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

Evidence of a novel cross‐species transmission by ovine papillomaviruses

Falco

Cutarelli

Cuccaro

et al. 2022

Transbounding Emerging Dis

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“…Given that our patients were mainly adults, this difference could be due to quantitative differences of fungal cells in the blood between adults and neonates after the onset of disease (sampling time). The ddPCR method also has other diagnostic advantages over conventional qPCR, including lower sample volume requirements and faster execution times, because the thermocycling times are shorter ( De falco et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

Chen

Ying-guang²,

Zhang³

et al. 2021

Front. Microbiol.

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Numerous studies have shown that droplet digital PCR (ddPCR) is a promising tool for the diagnosis of pathogens, especially in samples with low concentrations of pathogenic DNA. An early diagnosis of candidemia is critical for the effective treatment of patients. In this study, we evaluated the sensitivity and specificity of ddPCR assay for Candida DNA detection both in vitro by mixing fungal cells with human blood and in vivo by analyzing blood samples from infected mice and patients with suspected candidemia. The results showed that ddPCR assay could detect a minimum of 4.5 DNA copies per reaction in blood samples. ddPCR showed higher sensitivity and specificity for Candida DNA detection than traditional culture and quantitative PCR (qPCR) methods and also exhibited significantly better positive and negative predictive values than the culture and qPCR methods that were commonly used in clinical practice. Hence, our study demonstrates that ddPCR assay is a promising method for the timely diagnosis of candidemia and could be useful for monitoring the treatment of candidemia.

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Digital droplet PCR for the detection and quantification of circulating bovine Deltapapillomavirus

Cited by 2 publications

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Evidence of a novel cross‐species transmission by ovine papillomaviruses

Evidence of a novel cross‐species transmission by ovine papillomaviruses

Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

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