scholarly journals Detection and Quantification of Bovine Papillomavirus DNA by Digital Droplet PCR in Sheep Blood

2021 ◽  
Author(s):  
Sante Roperto ◽  
Anna Cutarelli ◽  
Federica Corrado ◽  
Francesca De Falco ◽  
Canio Buonavoglia
Keyword(s):  
Animal Husbandry ◽  
Bovine Papillomavirus ◽  
Liquid Biopsies ◽  
Highly Pathogenic ◽  
Blood Samples ◽  
Healthy Sheep ◽  
Droplet Pcr ◽  
First Time ◽  
Papillomavirus Dna ◽  
Detection And Quantification

Abstract Highly pathogenic bovine Delta papillomaviruses (δPVs) were detected and quantified for the first time using digital droplet polymerase chain reaction (ddPCR) by liquid biopsy in 103 clinically healthy sheep. Overall, ddPCR detected bovine δPVs in 68 blood samples (66%). Bovine papillomavirus (BPV) infection by a single genotype was revealed in 59% of the blood samples, and BPV coinfection by double, triple or quadruple genotypes was observed in 41% of liquid biopsies. The BPV-2 genotype was most frequently seen in sheep, whereas BPV-1 was the least common. Furthermore, ddPCR was very useful for detection and quantification; the BPV-14 genotype was observed for the first time in ovine species, displaying the highest prevalence in some geographical areas (Apulia). In 42 of the positive samples (61.8%), a single BPV infection was observed, 26 of which were caused by BPV-2 (61.9%) and 7 by BPV-13 (16.7%). BPV-14 was responsible for 7 single infections (16.7%) and BPV-1 for 2 single infections (4.7%). Multiple BPV coinfections were observed in the remaining 26 positive samples (38.2%), with dual BPV-2/BPV-13 infection being the most prevalent (84.6%). BPV infection by triple and quadruple genotypes was also observed in 11.5% and 3.8% of cases, respectively. The present study showed that ddPCR, a biotechnological refinement of conventional PCR, is by far the most sensitive and accurate assay for BPV detection. Therefore, ddPCR displayed diagnostic and epidemiological value resulting in the identification of otherwise undetectable BPV genotypes as well as their geographical distributions and suggesting that animal husbandry practices contribute to cross-species transmission of BPVs..

scholarly journals Detection and quantification of bovine papillomavirus DNA by digital droplet PCR in sheep blood

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sante Roperto ◽  
Anna Cutarelli ◽  
Federica Corrado ◽  
Francesca De Falco ◽  
Canio Buonavoglia
Keyword(s):  
Animal Husbandry ◽  
Bovine Papillomavirus ◽  
Liquid Biopsies ◽  
Highly Pathogenic ◽  
Blood Samples ◽  
Healthy Sheep ◽  
Droplet Pcr ◽  
First Time ◽  
Papillomavirus Dna ◽  
Detection And Quantification

AbstractHighly pathogenic bovine papillomaviruses (BPVs) were detected and quantified for the first time using digital droplet polymerase chain reaction (ddPCR) by liquid biopsy in 103 clinically healthy sheep. Overall, ddPCR detected BPVs in 68 blood samples (66%). BPV infection by a single genotype was revealed in 61.8% of the blood samples, and BPV coinfection by double, triple or quadruple genotypes was observed in 38.2% of liquid biopsies. The BPV-2 genotype was most frequently seen in sheep, whereas BPV-1 was the least common. Furthermore, ddPCR was very useful for detection and quantification; the BPV-14 genotype was observed for the first time in ovine species, displaying the highest prevalence in some geographical areas (Apulia). In 42 of the positive samples (61.8%), a single BPV infection was observed, 26 of which were caused by BPV-2 (61.9%) and 7 by BPV-13 (16.7%). BPV-14 was responsible for 7 single infections (16.7%) and BPV-1 for 2 single infections (4.7%). Multiple BPV coinfections were observed in the remaining 26 positive samples (38.2%), with dual BPV-2/BPV-13 infection being the most prevalent (84.6%). BPV infection by triple and quadruple genotypes was also observed in 11.5% and 3.8% of cases, respectively. The present study showed that ddPCR, a biotechnological refinement of conventional PCR, is by far the most sensitive and accurate assay for BPV detection compared to conventional qPCR. Therefore, ddPCR displayed an essential diagnostic and epidemiological value very useful for the identification of otherwise undetectable BPV genotypes as well as their geographical distributions and suggesting that animal husbandry practices contribute to cross-species transmission of BPVs.

scholarly journals Molecular Epidemiology of Ovine Papillomavirus Infections Among Sheep in Southern Italy

2021 ◽  
Vol 8 ◽  
Author(s):  
Francesca De Falco ◽  
Anna Cutarelli ◽  
Nicola D'Alessio ◽  
Pellegrino Cerino ◽  
Cornel Catoi ◽  
...  
Keyword(s):  
Molecular Epidemiology ◽  
Real Time ◽  
Molecular Diagnostic ◽  
Multiple Infections ◽  
Blood Samples ◽  
Real Time Quantitative Pcr ◽  
Healthy Sheep ◽  
Digital Polymerase Chain Reaction ◽  
First Time ◽  
Real Time Qpcr

Ovine papillomaviruses (OaPVs) were detected and quantified, for the first time, using droplet digital polymerase chain reaction (ddPCR) and real-time quantitative PCR (qPCR) via blood samples of 165 clinically healthy sheep. OaPV DNA was detected in 126 blood samples (~76.4%). DdPCR detected OaPV DNA in 124 samples; in only two additional samples positive for real-time qPCR, ddPCR failed to detect the presence of any OaPVs. In 70 of the positive samples (~55.6%), a single OaPV infection was observed, 12 of which were caused by OaPV1 (~17.1%) and 14 by OaPV2 (20%). OaPV3 was responsible for 19 single infections (~27.1%), and OaPV4 for 25 single infections (~35.7%). Multiple OaPV coinfections were observed in 56 (~44.4%) positive samples. OaPV coinfections caused by two genotypes were observed in 31 positive samples (~55.4%), with dual OaPV3/OaPV4 infection being the most prevalent as seen in 11 blood samples. In addition, five OaPV1/OaPV4, four OaPV1/OaPV2, four OaPV2/OaPV3, four OaPV1/OaPV3, and three OaPV2/OaPV4 dual coinfections were also detected. OaPV coinfections by triple and quadruple genotypes were detected in 24 (~42.8%) and only one (~1.8%) of coinfected blood samples, respectively. Multiple infections caused by OaPV1/OaPV3/OaPV4 genotypes were the most prevalent, as observed in 12 (50%) blood samples harboring triple OaPV infections. This study showed that ddPCR is the most sensitive and accurate assay for OaPV detection and quantification thus outperforming real-time qPCR in terms of sensitivity and specificity. Therefore, ddPCR may represent the molecular diagnostic tool of choice, ultimately providing useful insights into OaPV molecular epidemiology and field surveillance.

Monitoring clinical levels of heparin in human blood samples with an indicator-displacement assay

2015 ◽  
Vol 51 (10) ◽  
pp. 1953-1956 ◽  
Author(s):  
Jean-Patrick Francoia ◽  
Robert Pascal ◽  
Laurent Vial
Keyword(s):  
Human Blood ◽  
Fluorescent Detection ◽  
Blood Samples ◽  
Turn On ◽  
Displacement Assay ◽  
Human Blood Samples ◽  
First Time ◽  
Detection And Quantification

Herein, we report a simple assay – involving the displacement of a new designed dye from a dendrigraft surface – that allows for the first time the turn-ON fluorescent detection and quantification of heparin in human blood at clinically relevant levels.

Detection and Quantification of 7-Hydroxydehydroepiandrosterone Epimers in Three Body Fluids

2002 ◽  
Vol 67 (1) ◽  
pp. 10-18 ◽  
Author(s):  
Richard Hampl ◽  
Martin Hill ◽  
Luboslav Stárka
Keyword(s):  
Healthy Subjects ◽  
Serum Levels ◽  
Body Fluids ◽  
Gas Chromatography Mass Spectrometry ◽  
Specific Radioimmunoassay ◽  
Order Of Magnitude ◽  
Three Body ◽  
First Time ◽  
Nanomolar Range ◽  
Detection And Quantification

3β,7α-Dihydroxyandrost-5-en-17-one (1) (7α-OH-DHEA) and its 7β-hydroxy epimer 2 (7β-OH-DHEA) - 7α- and 7β-hydroxydehydroepiandrosterone - were detected and quantified in three human body fluids: in blood serum, saliva and ejaculate. Specific radioimmunoassay and gas chromatography-mass spectrometry have been used. For the first time the data on changes of these dehydroepiandrosterone metabolites are reported for a representative group of healthy subjects of both sexes (172 females and 217 males) during the life span. The serum levels of both 7-hydroxydehydroepiandrosterone epimers in serum and also in semen were in the low nanomolar range, while concentrations by one order of magnitude lower were found in saliva, but still within the detection limit. The results will serve as a basis for comparative studies of 7-hydroxydehydroepiandrosterone levels under various pathophysiological conditions, with a particular respect to autoimmune disorders.

Vanadate enhances transformation of bovine papillomavirus DNA-transfected C3H/10T1/2 cells

1992 ◽  
Vol 64 (1) ◽  
pp. 83-90 ◽  
Author(s):  
Linda Anne Kowalski ◽  
Su-Sing Tsang ◽  
Allan J. Davison
Keyword(s):  
Bovine Papillomavirus ◽  
Papillomavirus Dna

Enhanced production of hepatitis B surface antigen in NIH 3T3 mouse fibroblasts by using extrachromosomally replicating bovine papillomavirus vector

1983 ◽  
Vol 3 (6) ◽  
pp. 1032-1039
Author(s):  
Y Wang ◽  
C Stratowa ◽  
M Schaefer-Ridder ◽  
J Doehmer ◽  
P H Hofschneider
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Hepatitis B Surface Antigen ◽  
Bovine Papillomavirus ◽  
Mouse Fibroblasts ◽  
Antigen Gene ◽  
B Virus ◽  
Nih 3T3 ◽  
Papillomavirus Dna

We have constructed a recombinant pBR322 plasmid composed of a subgenomic transforming fragment of bovine papillomavirus DNA and the hepatitis B surface antigen gene from cloned hepatitis B virus DNA and used it for transfection of NIH 3T3 mouse fibroblasts. The transformed cells retain the plasmids in extrachromosomal form with a copy number of about 50 to 100 per cell. Expression of the hepatitis B surface antigen gene linked to bovine papillomavirus DNA is independent of its orientation relative to the bovine papillomavirus vector. Cell lines continuously secreting high amounts of hepatitis B surface antigen into the medium could be established. The antigen is released into the culture medium as 22-nm particles, having the same physical properties and constituent polypeptides as those found in the serum of hepatitis B virus-infected patients.

Airborne Alt a 1 Dynamic and Its Relationship With the Airborne Dynamics of Alternaria Conidia and Pleosporales Spores

2021 ◽  
Author(s):  
Concepción De Linares ◽  
David Navarro ◽  
Rut Puigdemunt ◽  
Jordina Belmonte
Keyword(s):  
Public Health Service ◽  
Fungal Spores ◽  
High Volume ◽  
Major Allergen ◽  
Allergic Reactions ◽  
Aerobiological Monitoring ◽  
Treatment And Prevention ◽  
First Time ◽  
Prevention Of Allergy ◽  
Detection And Quantification

Fungal spores are universal atmospheric components associated to allergic reactions. Alternaria (Ascomycota) is considered the most allergenic spore taxa. Alt a 1 is the major allergen of Alternaria and is present also in other Pleosporales. In this study, standard Hirst-based sampling and analyzing methods for measuring spore daily concentrations of Alternaria, Curvularia, Drechslera-Helminthosporium, Epicoccum, Leptosphaeria, Pithomyces, Pleospora and Stemphyllium (all included in the taxon Pleosporales) have been used besides two high-volume samplers, Burkard Cyclone (2017) and MCV CAV-A/mb (2019-2020), and ELISA Kits for measuring the allergen. The detection and quantification of Alt a 1 was only possible in the samples from the MCV sampler. Although Alt a 1 was better correlated with Alternaria spores than with Pleosporales spores, the three of them showed high correlations. It is shown, for the first time, a high and significant correlation of Alt a 1 with temperature, a negative one with relative humidity and no correlation with precipitation. The aerobiological monitoring of these three elements ensures the best information for understanding the affectation to allergy sufferers but, if not possible, as a minimum public health service aiming at the detection, treatment and prevention of allergy, the study of the airborne Alternaria spores should be ensured.

scholarly journals Detection and characterisation of papillomavirus in skin lesions of giraffe and sable antelope in South Africa

2011 ◽  
Vol 82 (2) ◽  
Author(s):  
E. Van Dyk ◽  
A-M Bosman ◽  
E. Van Wilpe ◽  
J. H. Williams ◽  
R. G. Bengis ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Skin Lesions ◽  
Immunoperoxidase Staining ◽  
Bovine Papillomavirus ◽  
Virus Particles ◽  
Tissue Sections ◽  
Equine Sarcoid ◽  
Pcr Product ◽  
Papillomavirus Dna

Papillomavirus was detected electron microscopically in cutaneous fibropapillomas of a giraffe (Giraffa camelopardalis) and a sable antelope (Hippotragus niger). The virus particles measured 45 nm in diameter. Histopathologically, the lesions showed histopathological features similar to those of equine sarcoid as well as positive immunoperoxidase-staining of tissue sections for papillomavirus antigen. Polymerase chain reaction (PCR) detected bovine papillomavirus (BPV) DNA. Bovine papillomavirus-1 was characterised by real-time PCR in the sable and giraffe, and cloning and sequencing of the PCR product revealed a similarity to BPV-1. As in the 1st giraffe, the lesions from a 2nd giraffe revealed locally malignant pleomorphism, possibly indicating the lesional end-point of papilloma infection. Neither virus particles nor positively staining papillomavirus antigen could be demonstrated in the 2nd giraffe but papillomavirus DNA was detected by real-time PCR which corresponded with BPV-1 and BPV-2.

Spontaneous Cutaneous Papillomatosis in Yaks and Detection and Quantification of Bovine Papillomavirus-1 and -2

2012 ◽  
Vol 60 (5) ◽  
pp. 475-480 ◽  
Author(s):  
J. Bam ◽  
P. Kumar ◽  
G. D. Leishangthem ◽  
A. Saikia ◽  
R. Somvanshi
Keyword(s):  
Bovine Papillomavirus ◽  
Detection And Quantification

scholarly journals Prevalence of Anti-Phospholipid Autoantibodies and Their Association with Respiratory SOFA Component in Patients with COVID-19 Pneumonia: A Prospective Cohort Analysis in Tunisia

2021 ◽  
Author(s):  
Amal ABOUDA ◽  
Yasmine BOUKHALFA ◽  
Wafa ANENE ◽  
Zied HAJJEJ ◽  
Ezzeddine GHAZOUANI ◽  
...  
Keyword(s):  
Critically Ill ◽  
Significant Positive Correlation ◽  
Cohort Analysis ◽  
Observational Cohort Study ◽  
Prospective Observational Cohort Study ◽  
Respiratory Organ ◽  
Blood Samples ◽  
Observational Cohort ◽  
First Time ◽  
Independent Indicator

Abstract Purpose: The aim of our study was to evaluate the prevalence of aPLAs among Tunisian critically-ill covid19 and non-covid19 patients and to investigate the clinical significance of aPLAs by determining the SOFA score and their respiratory failure during their ICU stay. Methods: We conducted a prospective observational cohort study including critically ill COVID-19 patients and non-COVID-19 patients with pulmonary origin sepsis, admitted to the intensive care unit. Blood samples were collected on days 1, 3, 5, 8 and 10 of hospitalization in order to measure titers of anti-cardiolipin (aCL), anti-phosphatidylserine (aPS) by chemiluminescence immunoassay. Results: We enrolled 43 COVID-19 patients and 31 non COVID-19 with pulmonary origin sepsis. In-hospital mortality rate was significantly higher (p=0.026) in COVID-19 patients (79%). 58.8% of COVID-19 patients were aPLA positive; however, only 22.5% of the non-COVID-19 were positive for aPLA (p=0.002). A significant positive correlation existed between respiratory SOFA component at days 3, 5, 8 and 10 and anti-phospholipid antibodies concentrations. Conclusion: Based on our results, for the first time, anti-phospholipid antibodies may be used as an independent indicator of respiratory organ failure in critically ill patients, to stratify and assess the prognosis of pulmonary origin sepsis and COVID-19.

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