scholarly journals Canine distemper virus detection in asymptomatic and non vaccinated dogs

2010 ◽  
Vol 30 (2) ◽  
pp. 139-144 ◽  
Author(s):  
Helen L. Del Puerto ◽  
Anilton C. Vasconcelos ◽  
Luciana Moro ◽  
Fabiana Alves ◽  
Gissandra F. Braz ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Canine Distemper Virus ◽  
Early Stage ◽  
Virus Detection ◽  
Vero Cells ◽  
Canine Distemper ◽  
Coding Region ◽  
Protein Coding ◽  
Dissociation Curves

A quantitative real time polymerase chain reaction (PCR) revealed canine distemper virus presence in peripheral blood samples from asymptomatic and non vaccinated dogs. Samples from eleven domestic dogs with no signs of canine distemper and not vaccinated at the month of collection were used. Canine distemper virus vaccine samples in VERO cells were used as positive controls. RNA was isolated with Trizol®, and treated with a TURBO DNA-free kit. Primers were designed for canine distemper virus nucleocapsid protein coding region fragment amplification (84 bp). Canine b-actin (93 bp) was utilized as the endogenous control for normalization. Quantitative results of real time PCR generated by ABI Prism 7000 SDS Software showed that 54.5% of dogs with asymptomatic canine distemper were positive for canine distemper virus. Dissociation curves confirmed the specificity of the real time PCR fragments. This technique could detect even a few copies of viral RNA and identificate subclinically infected dogs providing accurate diagnosis of this disease at an early stage.

scholarly journals Detection and differentiation of field and vaccine strains of canine distemper virus using reverse transcription followed by nested real time PCR (RT-nqPCR) and RFLP analysis

2013 ◽  
Vol 194 (1-2) ◽  
pp. 39-45 ◽  
Author(s):  
Cristine Dossin Bastos Fischer ◽  
Nilo Ikuta ◽  
Cláudio Wageck Canal ◽  
Aline Makiejczuk ◽  
Mariangela da Costa Allgayer ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Canine Distemper Virus ◽  
Rflp Analysis ◽  
Canine Distemper

TaqMan Based Real Time PCR for the Quantification of Canine Distemper Virus

2007 ◽  
Vol 31 (S1) ◽  
pp. 261-263 ◽  
Author(s):  
A. Scagliarini ◽  
F. Dal Pozzo ◽  
L. Gallina ◽  
F. Vaccari ◽  
L. Morganti
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Canine Distemper Virus ◽  
Canine Distemper

Strip-dried blood sampling: applicability for bovine leukemia virus detection with ELISA and real-time PCR

2019 ◽  
Vol 263 ◽  
pp. 101-104 ◽  
Author(s):  
Nikolay Yu. Saushkin ◽  
Jeanne V. Samsonova ◽  
Alexander P. Osipov ◽  
Sergey E. Kondakov
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Bovine Leukemia Virus ◽  
Leukemia Virus ◽  
Virus Detection ◽  
Blood Sampling

Canine Distemper Virus Induces Apoptosis Through Caspase-3 and -8 Activation in Vero Cells

2006 ◽  
Vol 53 (6) ◽  
pp. 273-277 ◽  
Author(s):  
M. Kajita ◽  
H. Katayama ◽  
T. Murata ◽  
C. Kai ◽  
M. Hori ◽  
...  
Keyword(s):  
Canine Distemper Virus ◽  
Caspase 3 ◽  
Vero Cells ◽  
Canine Distemper

Inhibition of canine distemper virus replication by blocking pyrimidine nucleotide synthesis with A77 1726, the active metabolite of the anti-inflammatory drug leflunomide

2021 ◽  
Author(s):  
Yao Li ◽  
Li Yi ◽  
Sipeng Cheng ◽  
Yongshan Wang ◽  
Jiongjiong Wang ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Active Metabolite ◽  
Canine Distemper Virus ◽  
De Novo ◽  
Specific Inhibitor ◽  
Vero Cells ◽  
Canine Distemper ◽  
Nucleotide Synthesis ◽  
Pyrimidine Nucleotide ◽  
Rate Limiting

Canine distemper virus (CDV) is the aetiological agent that causes canine distemper (CD). Currently, no antiviral drugs have been approved for CD treatment. A77 1726 is the active metabolite of the anti-rheumatoid arthritis (RA) drug leflunomide. It inhibits the activity of Janus kinases (JAKs) and dihydroorotate dehydrogenase (DHO-DHase), a rate-limiting enzyme in de novo pyrimidine nucleotide synthesis. A77 1726 also inhibits the activity of p70 S6 kinase (S6K1), a serine/threonine kinase that phosphorylates and activates carbamoyl-phosphate synthetase (CAD), a second rate-limiting enzyme in the de novo pathway of pyrimidine nucleotide synthesis. Our present study focuses on the ability of A77 1726 to inhibit CDV replication and its underlying mechanisms. Here we report that A77 1726 decreased the levels of the N and M proteins of CDV and lowered the virus titres in the conditioned media of CDV-infected Vero cells. CDV replication was not inhibited by Ruxolitinib (Rux), a JAK-specific inhibitor, but by brequinar sodium (BQR), a DHO-DHase-specific inhibitor, and PF-4708671, an S6K1-specific inhibitor. Addition of exogenous uridine, which restores intracellular pyrimidine nucleotide levels, blocked the antiviral activity of A77 1726, BQR and PF-4708671. A77 1726 and PF-4708671 inhibited the activity of S6K1 in CDV-infected Vero cells, as evidenced by the decreased levels of CAD and S6 phosphorylation. S6K1 knockdown suppressed CDV replication and enhanced the antiviral activity of A77 1726. These observations collectively suggest that the antiviral activity of A77 1726 against CDV is mediated by targeting pyrimidine nucleotide synthesis via inhibiting DHO-DHase activity and S6K1-mediated CAD activation.

scholarly journals Gene expression patterns in bone following lipopolysaccharide stimulation

2014 ◽  
Vol 19 (4) ◽  
Author(s):  
Jing Yang ◽  
Nan Su ◽  
Xiaolan Du ◽  
Lin Chen
Keyword(s):  
Gene Expression ◽  
Bone Formation ◽  
Real Time ◽  
Real Time Pcr ◽  
Early Stage ◽  
System Development ◽  
Expression Patterns ◽  
Osteoclast Differentiation ◽  
Gene Expression Patterns ◽  
Expression Levels

AbstractBone displays suppressed osteogenesis in inflammatory diseases such as sepsis and rheumatoid arthritis. However, the underlying mechanisms have not yet been clearly explained. To identify the gene expression patterns in the bone, we performed Affymetrix Mouse Genome 430 2.0 Array with RNA isolated from mouse femurs 4 h after lipopolysaccharide (LPS) administration. The gene expressions were confirmed with real-time PCR. The serum concentration of the N-terminal propeptide of type I collagen (PINP), a bone-formation marker, was determined using ELISA. A total of 1003 transcripts were upregulated and 159 transcripts were downregulated (more than twofold upregulation or downregulation). Increased expression levels of the inflammation-related genes interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) were confirmed from in the period 4 h to 72 h after LPS administration using real-time PCR. Gene ontogene analysis found four bone-related categories involved in four biological processes: system development, osteoclast differentiation, ossification and bone development. These processes involved 25 upregulated genes. In the KEGG database, we further analyzed the transforming growth factor β (TGF-β) pathway, which is strongly related to osteogenesis. The upregulated bone morphogenetic protein 2 (BMP2) and downregulated inhibitor of DNA binding 4 (Id4) expressions were further confirmed by real-time PCR after LPS stimulation. The osteoblast function was determined through examination of the expression levels of core binding factor 1 (Cbfa1) and osteocalcin (OC) in bone tissues and serum PINP from 4 h to 72 h after LPS administration. The expressions of OC and Cbfa1 decreased 6 h after administration (p < 0.05). Significantly suppressed PINP levels were observed in the later stage (from 8 h to 72 h, p < 0.05) but not in the early stage (4 h or 6 h, p > 0.05) of LPS stimulation. The results of this study suggest that LPS induces elevated expressions of skeletal system development- and osteoclast differentiation-related genes and inflammation genes at an early stage in the bone. The perturbed functions of these two groups of genes may lead to a faint change in osteogenesis at an early stage of LPS stimulation. Suppressed bone formation was found at later stages in response to LPS stimulation.

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