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.

scholarly journals Peripheral blood mononuclear cells represent a reservoir of bovine papillomavirus DNA in sarcoid-affected equines

2008 ◽  
Vol 89 (6) ◽  
pp. 1390-1395 ◽  
Author(s):  
Sabine Brandt ◽  
Rhea Haralambus ◽  
Angelika Schoster ◽  
Reinhard Kirnbauer ◽  
Christian Stanek
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Virus Transmission ◽  
Skin Lesions ◽  
Host Cells ◽  
Bovine Papillomavirus ◽  
Peripheral Blood Mononuclear ◽  
Virus Latency ◽  
Equine Sarcoid ◽  
Papillomavirus Dna

Bovine papillomaviruses of types 1 and 2 (BPV-1 and -2) chiefly contribute to equine sarcoid pathogenesis. However, the mode of virus transmission and the presence of latent infections are largely unknown. This study established a PCR protocol allowing detection of ≤10 copies of the BPV-1/-2 genes E5 and L1. Subsequent screening of peripheral blood mononuclear cell (PBMC) DNA derived from horses with and without BPV-1/2-induced skin lesions demonstrated the exclusive presence of E5, but not L1, in PBMCs of BPV-1/2-infected equines. To validate this result, a blind PCR was performed from enciphered PBMC DNA derived from 66 horses, revealing E5 in the PBMCs of three individuals with confirmed sarcoids, whereas the remaining 63 sarcoid-free animals were negative for this gene. L1 could not be detected in any PBMC DNA, suggesting either deletion or interruption of this gene in PBMCs of BPV-1/-2-infected equines. These results support the hypothesis that PBMCs may serve as host cells for BPV-1/-2 DNA and contribute to virus latency.

scholarly journals Co-infection of bovine papillomavirus type-1 and -10 in teat warts of dairy cattle

2013 ◽  
Vol 58 (No. 12) ◽  
pp. 605-608
Author(s):  
P. Kumar ◽  
BL Jangir ◽  
G. Saikumar ◽  
R. Somvanshi
Keyword(s):  
Dairy Cattle ◽  
Real Time ◽  
Real Time Pcr ◽  
Rice Grain ◽  
Bovine Papillomavirus ◽  
Viral Dna ◽  
Specific Primers ◽  
Copy Numbers ◽  
Pcr Techniques

The present study was carried out to investigate the involvement of different bovine papillomaviruses in the teat warts of cattle. A total of 11 teat wart samples showing rice grain-like and small, sessile elevated greyish or flesh-like growths were collected from dairy cattle. DNA was extracted from these teat wart samples and PCR and real time PCR techniques were applied using specific primers for BPV-1 and -10 to detect the presence of viral nucleic acid. PCR revealed the presence of viral DNA of BPV-1 and -10 in three and seven samples, respectively. Quantification using real time PCR revealed that the copy numbers of the viral DNA of BPV-1 and -10 DNA varied from 1.12E + 04 to 2.99E + 04 and 3.56E + 02 to 5.23E + 06, respectively. From the present study it can be concluded that BPV-1 and -10 are involved in production of rice grain-like and sessile elevated growths on the teats of cattle.

Bovine papillomavirus DNA can be detected in keratinocytes of equine sarcoid tumors

2010 ◽  
Vol 146 (3-4) ◽  
pp. 269-275 ◽  
Author(s):  
Lies Bogaert ◽  
Ann Martens ◽  
Wijbe Martin Kast ◽  
Eric Van Marck ◽  
Hilde De Cock
Keyword(s):  
Bovine Papillomavirus ◽  
Equine Sarcoid ◽  
Papillomavirus Dna

scholarly journals Clinical Validation of the HPV-Risk Assay, a Novel Real-Time PCR Assay for Detection of High-Risk Human Papillomavirus DNA by Targeting the E7 Region

2014 ◽  
Vol 52 (3) ◽  
pp. 890-896 ◽  
Author(s):  
A. T. Hesselink ◽  
J. Berkhof ◽  
M. L. van der Salm ◽  
A. P. van Splunter ◽  
T. H. Geelen ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
High Risk ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Validation ◽  
Pcr Assay ◽  
Papillomavirus Dna

Intralesional bovine papillomavirus DNA loads reflect severity of equine sarcoid disease

2010 ◽  
Vol 42 (4) ◽  
pp. 327-331 ◽  
Author(s):  
R. HARALAMBUS ◽  
J. BURGSTALLER ◽  
J. KLUKOWSKA-RÖTZLER ◽  
R. STEINBORN ◽  
S. BUCHINGER ◽  
...  
Keyword(s):  
Bovine Papillomavirus ◽  
Equine Sarcoid ◽  
Papillomavirus Dna

scholarly journals Comparative evaluation of published cytomegalovirus primers for rapid real-time PCR: which are the most sensitive?

2009 ◽  
Vol 58 (7) ◽  
pp. 878-883 ◽  
Author(s):  
Wafa Habbal ◽  
Fawza Monem ◽  
Barbara C. Gärtner
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Analytical Sensitivity ◽  
Pcr Product ◽  
Highly Sensitive ◽  
Primer Sets ◽  
Analytical Sensitivities ◽  
Strain Ad169

Standardization of human cytomegalovirus (CMV) PCR is highly recommended. As primer design is essential for PCR sensitivity, this study evaluated all published CMV primer pairs to identify the most sensitive for single-round real-time PCR. PubMed (1993–2004) was searched for original papers aimed at CMV PCR. Fifty-seven papers were identified revealing 82 different primer pairs. Of these, 17 primer sets were selected for empirical study, as they were either used in real-time PCR or were evaluated comparatively by conventional PCR. After optimizing the PCR conditions, these primer sets were evaluated by real-time PCR using a SYBR Green format. Analytical sensitivities were assessed by testing the reference standard CMV strain AD169. A blast search was performed to identify mismatches with published sequences. Additionally, 60 clinical samples were tested with the three primer sets showing highest analytical sensitivity and the best match to all CMV strains. Three primer sets located in the glycoprotein B (UL55) gene region were found to be the most sensitive using strain AD169. However, two of these showed a considerable number of mismatches with clinical isolates in a blast search. Instead, two other pairs from the lower matrix phosphoprotein (UL83) gene and DNA polymerase (UL54) gene showed reasonable sensitivity and no mismatches with clinical isolates. These three pairs were further tested with clinical samples, which indicated that the two primer sets from UL55 and UL54 were the most sensitive. Interestingly, the analytical sensitivity of the PCR was inversely correlated with the size of the PCR product. In conclusion, these two primer pairs are recommended for a standardized, highly sensitive, real-time PCR.

Real-Time Polymerase Chain Reaction Detection of Bovine DNA in Meat and Bone Meal Samples

2002 ◽  
Vol 65 (7) ◽  
pp. 1158-1165 ◽  
Author(s):  
S. LAHIFF ◽  
M. GLENNON ◽  
J. LYNG ◽  
T. SMITH ◽  
N. SHILTON ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat And Bone Meal ◽  
Bone Meal ◽  
Chain Reaction ◽  
Pcr Assay ◽  
The Real ◽  
Pcr Product ◽  
Pcr Products ◽  
Polymerase Chain

We describe a real-time polymerase chain reaction (PCR) assay for the detection of bovine DNA extracted from meat and bone meal (MBM) samples. PCR primers were used to amplify a 271-bp region of the mitochondrial ATPase 8–ATPase 6 gene, and a fluorogenic probe (BOV1) labeled with a 5′ FAM reporter and a 3′ TAMRA quencher was designed to specifically detect bovine PCR product. The specificity of the BOV1 probe for the detection of the bovine PCR product was confirmed by Southern blot hybridization analysis of the probe with PCR products generated from ovine, porcine, and bovine genomic DNA extracted from blood and with PCR products generated from genomic DNA extracted from single-species laboratory scale rendered MBM samples. The specificity of the BOV1 probe was also evaluated in real-time PCR reactions including these genomic targets. Both methods demonstrated that the BOV1 probe was specific for the detection of bovine PCR product. The BOV1 probe had a detection limit of 0.0001% bovine material by Southern blot DNA probe hybridization analysis and a detection limit of 0.001% bovine material in the real-time PCR assay. Application of the real-time PCR assay to six industrial samples that had previously tested positive for the presence of bovine material with a conventional PCR assay yielded positive results with the real-time PCR assay for four samples.

Molecular Gene Expression of Toll-Like Receptors 4 & 10 in Cellular Subsets of Human Peripheral Blood among Patients with Prostatitis: Conventional, Real Time Pcr and DNA Sequencing Techniques

2020 ◽  
Vol 7 (11) ◽  
pp. 5103-5110
Author(s):  
Ihsan AlSaimary1 ◽  
Hussein Aldhaheri ◽  
Murtadha ALMusafer2
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Human Peripheral Blood ◽  
Receptor Gene ◽  
Gene Clusters ◽  
Control Group ◽  
Agarose Gel ◽  
Toll Like Receptor ◽  
Blood Samples ◽  
Pcr Product

The Aim of this study was to determine Immunogenetic expression of  Toll-like receptor gene clusters related to prostatitis, to give acknowledge about Role of TLR in prostatitis immunity in men from Basrah and Maysan provinces. A case–control study included 135 confirmed prostatitis patients And 50 persons as a control group. Data about age, marital status, working, infertility, family history and personal information like (Infection, Allergy, Steroid therapy, Residency, Smoking, Alcohol Drinking, Blood group, Body max index (BMI) and the clinical finding for all patients of Prostatitis were collected , The molecular expression study include extracting DNA from blood of Prostatitis patients , Prostitis patients and Control group by using specific primers for conventional PCR and Real Time PCR , the amplification of all extracted DNA from blood samples was preform and confirm by using electrophoresis with (100volt/30min).  The amplification of all extracted DNA from blood samples was preform and confirm by using electrophoresis with (100volt/30min) , the result of this estimation revealed that the amplified DNA(PCR product) was 227bp for TLR4 on agarose gel (1%) , (50voltage for 1hour ) with a presence 100% ,  (PCR product) was 279bp for TLR10 on agarose gel(1%) , (50volt/1hour) with a presence 80%. The results of the present study indicate that the Toll like receptor alleles associated with risk of prostatitis.

scholarly journals Quantitative Detection of Trichloroethylene-Degrading Mycobacterium sp. TA27 with a Real-time PCR Product Detection System.

2001 ◽  
Vol 16 (2) ◽  
pp. 109-116 ◽  
Author(s):  
Akiko Hashimoto ◽  
Kazuhiro Iwasaki ◽  
Mutsuyasu Nakajima ◽  
Osami Yagi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Detection System ◽  
Quantitative Detection ◽  
Pcr Product

scholarly journals Changes in the microbiological diagnosis and epidemiology of cutaneous leishmaniasis in real-time PCR era: A six-year experience in a referral center in Barcelona

2021 ◽  
Vol 15 (11) ◽  
pp. e0009884
Author(s):  
Aroa Silgado ◽  
Mayuli Armas ◽  
Adrián Sánchez-Montalvá ◽  
Lidia Goterris ◽  
Maria Ubals ◽  
...  
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Real Time ◽  
Real Time Pcr ◽  
Skin Lesions ◽  
Tertiary Hospital ◽  
Molecular Techniques ◽  
Cutaneous Lesions ◽  
Imported Cases ◽  
Epidemiological Characteristics

Background Leishmaniasis is a neglected disease caused by different species of the protozoa Leishmania spp. Cutaneous lesions are the most common clinical manifestation. This disease is prevalent in tropical and subtropical areas, including the Mediterranean basin. In Spain, Leishmania (L.) infantum is the only endemic species, but imported cases are often diagnosed. Different classical parasitological methods can be performed for cutaneous leishmaniasis (CL) diagnosis; but currently molecular techniques serve as a relevant tool for the detection and characterization of Leishmania parasites. We aimed to evaluate clinical and epidemiological characteristics of CL diagnosed patients by real-time PCR in a tertiary hospital over a six-year period. Methodology/Principal findings Clinical, epidemiological and microbiological data were retrospectively collected and analyzed. In our study, CL was confirmed in 59 (31.4%) out of 188 patients by real-time PCR, showing an increase over recent years: 11 cases of CL between 2014 and 2016 and 48 between 2017 and 2019. Real-time PCR was performed on skin swabs and/or biopsies samples, with a positivity of 38.5% and 26.5%, respectively. Results were 100% concordant when biopsy and skin swab were performed simultaneously. L. (L.) infantum was the most frequent species detected (50%), followed by L. (L.) major (45%) and Viannia subgenus (5%), which were detected only in imported cases. L. (L.) major was almost entirely detected in travelers/migrants from Morocco. Multiple and atypical skin lesions were more common in imported cases than in autochthonous cases (44.4% vs. 21.8%). Conclusions/Significance An increase in both autochthonous and imported CL cases has been observed in past years in our hospital. Molecular techniques assist in improving CL diagnosis and characterization of the Leishmania species, mainly in imported cases.

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