scholarly journals Epidemiology of feline hemoplasmosis in the population of domestic cats in Istanbul

2021 ◽  
Vol 77 (02) ◽  
pp. 6490-2021
Author(s):  
BARAN CELIK ◽  
LORA KOENHEMSI ◽  
BANU DOKUZEYLUL ◽  
BEREN BAŞARAN KAHRAMAN ◽  
BELGI DIREN SIĞIRCI ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Statistical Significance ◽  
Strong Relationship ◽  
Epidemiological Data ◽  
Clinical Findings ◽  
Domestic Cats ◽  
First Time ◽  
The Relationship ◽  
Candidatus Mycoplasma Turicensis

The aim of this study was to determine the prevalence of hemoplasma species in cats by real-time PCR and to determine the distribution of the species. Furthermore, it was aimed to evaluate factors that are thought to be important in the epidemiology of the disease in cats statistically. For this purpose, blood samples from 246 cats were examined for Candidatus Mycoplasma haemominitum (CMhm), Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma turicensis (CMt) by real-time PCR. CMhm was detected in 20 (8.13%), CMt was detected in 3 (1.22%), and Mhf was found in 2 (0.81%) of 246 cats. At least one of the hemoplasma species was detected in 21 (8.54%) of 246 cats, and two different hemoplasma species were detected in 4 (19.05%) of 21 cats diagnosed with hemoplasma. The relationship between PCR positivity, the contact of cats with other cats (p = 0.02) and the detection of the intraoral wound (p = 0.001) was found to be statistically significant. The statistical significance of contact with other cats in the formation of the disease was revealed. The strong relationship between the presence of intraoral lesions and hemoplasma infection was revealed for the first time. Studies involving epidemiological data and their relationship with clinical findings should be continued.

scholarly journals Molecular Survey of Vector-Borne Pathogens of Dogs and Cats in Two Regions of Saudi Arabia

Pathogens ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 25
Author(s):  
Abdullah D. Alanazi ◽  
Abdulaziz S. Alouffi ◽  
Mohamed S. Alyousif ◽  
Mohammad Y. Alshahrani ◽  
Hend H. A. M. Abdullah ◽  
...  
Keyword(s):  
Saudi Arabia ◽  
Real Time ◽  
Real Time Pcr ◽  
Bartonella Henselae ◽  
Public Awareness ◽  
Effective Control ◽  
Vector Borne Diseases ◽  
Babesia Vogeli ◽  
Vector Borne ◽  
First Time

Dogs and cats play an important role as reservoirs of vector-borne pathogens, yet reports of canine and feline vector-borne diseases in Saudi Arabia are scarce. Blood samples were collected from 188 free-roaming dogs and cats in Asir (70 dogs and 44 cats) and Riyadh (74 dogs), Saudi Arabia. The presence of Anaplasma spp., Bartonella spp., hemotropic Mycoplasma spp., Babesia spp., and Hepatozoon spp. was detected using a multiplex tandem real-time PCR. PCR-positive samples were further examined with specific conventional and real-time PCR followed by sequencing. Dogs from Riyadh tested negative for all pathogens, while 46 out of 70 dogs (65.7%) and 17 out of 44 cats (38.6%) from Asir were positive for at least one pathogen. Positive dogs were infected with Anaplasma platys (57.1%), Babesia vogeli (30%), Mycoplasma haemocanis (15.7%), and Bartonella henselae (1.4%), and cats were infected with Mycoplasma haemofelis (13.6%), Candidatus Mycoplasma haemominutum (13.6%), B. henselae (9.2%), and A. platys (2.27%), all of which are reported for the first time in Saudi Arabia. Co-infection with A. platys and B. vogeli was detected in 17 dogs (24.28%), while coinfections were not detected in cats. These results suggest that effective control and public awareness strategies for minimizing infection in animals are necessary.

scholarly journals Building a molecular typing protocol for rs1801133 based on real-time PCR HRM technique

2019 ◽  
Vol 2 (3) ◽  
pp. 5-13
Author(s):  
Linh Thi Nhut Tran ◽  
My Thi Huynh Nguyen ◽  
Linh Nguy Hoang Le ◽  
Khoa Dang Le ◽  
Minh Hoang Nhat Nguyen ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Neurological Diseases ◽  
Human Diseases ◽  
Chromosome 1 ◽  
Nucleotide Sequencing ◽  
Sequencing Method ◽  
Mgcl2 Concentration ◽  
Clear Differentiation ◽  
The Relationship

rs1801133 is a single nucleotide polymorphism (SNP) located in the sequence of MTHFR on human chromosome 1. The alleles of this SNP affect the activity of the MTHFR enzyme. People bearing C/T genotype have 66% activity of MTHFR while people with T/T genotype have only 25% activity. These reduced activities of MTHFR cause homocysteinemia. There are several publications on the relationship between homocysteinemia and human diseases such as cardiovascular disease, neurological diseases, abnormal fetus, infertility and cancer. In this study, we built a molecular protocol for genotyping rs1801133 using real-time PCR HRM technique. This protocol could be used for diagnosis of molecular mechanism of homocysteinemia causing the mentoned above diseases as well as for the study of the relationship between rs1801133 and other human diseases. We successfully designed the primer pairs for genotyping and nucleotide sequencing rs1801133 by real-time PCR HRM and Sanger sequencing method. We also examined the optimal MgCl2 concentration for clear differentiation of three rs1801133 genotypes. Performance characteristics of the real-time PCR HRM protocol included of specificity, repeatability, reproducibility was evaluated and it showed good results. Comparison of genotyping results of rs1801133 between the realtime PCR HRM method and the Sanger nucleotide sequencing method showed good concordances. Finally, this real-time PCR HRM protocol for rs1801133 genotyping was applied on 100 human DNA samples to evaluate the clinical utility of the protocol.

scholarly journals Length of productive life the cows of Simmental breed

2021 ◽  
Vol 213 (10) ◽  
pp. 31-39
Author(s):  
L. Ignat'eva ◽  
A. Sermyagin
Keyword(s):  
Milk Yield ◽  
Russian Federation ◽  
Negative Relationship ◽  
Strong Relationship ◽  
Cattle Breeding ◽  
Productive Life ◽  
The Russian Federation ◽  
The Republic ◽  
First Time ◽  
The Relationship

Abstract. The purpose of the research was to assess the duration of the length of productive life of Simmental cows. Methods. The research was carried out on Simmental cows bred in 14 regions of the Russian Federation, the total livestock was 8 832 heads. The calculation of the heritability coefficients and correlation (genetic and paratypic) was carried out by using the programs RENUMF90 and REMLF90. Results. A fairly strong relationship was established between the duration of a productive life (months) and the age of culling (lactations) r = +0.795, the length of productive life (months) and lifetime productivity within the range of +0.669…+0.714. However, the relationship between the age of culling (lactations) and lifetime productivity is moderate, from +0.261 to +0.316. A moderate negative relationship was obtained between the age of culling (lactations) and milk yield per first lactation from –0.472 to –0.486. The average relationship was found between milk yield per first lactation and lifetime productivity from +0.567 to +0.588. Cows of the Altai Territory (3.08 lactations or 61.6 months), the Republic of Mordovia (3.38 lactations or 62.4 months) and the Lipetsk region (3.40 lactations or 65.7 months) were distinguished by low age of culling. While the greatest length of productive life was noted in animals and Bryansk (5.48 lactations or 86.9 months) and Irkutsk regions (4.57 lactations or 77.1 months). Bryansk (23 630 kg of milk), Tyumen (18 156 kg) and Irkutsk (17 751 kg) regions occupied the leading positions in lifetime productivity of cows in the sample, while the outsiders were the regions of traditional cattle breeding - Altai Territory (12658 kg of milk), the Republic of Bashkiria (12 482 kg). Scientific novelty. For the population Simmental cattle of the Russian Federation, for the first time, an assessment of selection and genetic parameters of lifelong productivity and length of productive life of Simmental cows was carried out, depending on the breeding region.

scholarly journals Polymorphism rs3087243 is associated with the occurrence of ankylosing spondylitis in the West Algerian population

2017 ◽  
Vol 1 (2) ◽  
pp. 29-30
Author(s):  
Chahinez Amira DAHMANI ◽  
Ahmed BENZAOUI ◽  
Fatima Zohra SEDIKI ◽  
Leila ADDA NEGGAZ ◽  
Faouzia ZEMANI FODIL ◽  
...  
Keyword(s):  
Ankylosing Spondylitis ◽  
Real Time ◽  
Real Time Pcr ◽  
Allele Frequencies ◽  
Healthy Individuals ◽  
The West ◽  
Algerian Population ◽  
Ctla4 Gene ◽  
First Time ◽  
Larger Sample

Background: Numerous studies have shown that polymorphism rs231775 of the CTLA4 gene is strongly implicated in the development of ankylosing spondylitis (AS). Other polymorphisms of this gene are candidates that may have an additional effect in susceptibility to AS. For the first time, we searched for the association of rs3087243 polymorphism located in the 3'UTR region of the CTLA4 gene with the development of SA in the Algerian population. Methods: The study involved 200 subjects (80 AS patients recruited at the rheumatology service and 120 healthy individuals unrelated). Genotyping was performed by real-time PCR (Taqman®). Analysis of the results was carried out by IBM.SPSS.Statictis® software. Results: The distribution of allele frequencies showed a significant association between the GG genotype of the polymorphism rs3087243 and AS risk (OR= 1.77 [0.98-3.21], p=0.004). Conclusion: Our data would suggest that the 3'UTR region of the CTLA4 gene could have an impact on the development of SA in the West Algerian population. These results need to be confirmed on a larger sample.

scholarly journals Real-time PCR for direct aptamer quantification on functionalized graphene surfaces

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Viviane C. F. dos Santos ◽  
Nathalie B. F. Almeida ◽  
Thiago A. S. L. de Sousa ◽  
Eduardo N. D. Araujo ◽  
Antero S. R. de Andrade ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Surface Modification ◽  
Silicon Dioxide ◽  
Real Time ◽  
Real Time Pcr ◽  
Functionalized Graphene ◽  
Graphene Surface ◽  
Precise Quantification ◽  
Graphene Functionalization ◽  
First Time

AbstractIn this study, we develop a real-time PCR strategy to directly detect and quantify DNA aptamers on functionalized graphene surfaces using a Staphylococcus aureus aptamer (SA20) as demonstration case. We show that real-time PCR allowed aptamer quantification in the range of 0.05 fg to 2.5 ng. Using this quantitative technique, it was possible to determine that graphene functionalization with amino modified SA20 (preceded by a graphene surface modification with thionine) was much more efficient than the process using SA20 with a pyrene modification. We also demonstrated that the functionalization methods investigated were selective to graphene as compared to bare silicon dioxide surfaces. The precise quantification of aptamers immobilized on graphene surface was performed for the first time by molecular biology techniques, introducing a novel methodology of wide application.

scholarly journals Decreased miRNA-146A in Glioblastoma Multiforme and Regulation of Cell Proliferation and Apoptosis by Target Notch1

2016 ◽  
Vol 31 (3) ◽  
pp. 270-275 ◽  
Author(s):  
Hong-qi Hu ◽  
Lai-guang Sun ◽  
Wu-jun Guo
Keyword(s):  
Glioblastoma Multiforme ◽  
Cell Viability ◽  
Real Time ◽  
Real Time Pcr ◽  
Glioma Cells ◽  
Viability Analysis ◽  
Nontumor Tissue ◽  
Proliferation And Apoptosis ◽  
A Cell ◽  
The Relationship

Objective The primary purpose of this paper is to investigate the relationship between the microRNA 146a (miR-146a) and the proliferation of cells occurring in glioblastoma multiforme. The secondary purpose of the paper is to investigate abnormalities of expression in miR-146a. Methods A real-time PCR assay was used to investigate the abnormal expression of miR-146a in glioma and adjacent tissue. Lipofection was used to transfect a mimic of miR-146a and induce the upregulation of miR-146a. Real-time PCR was used to observe the expression level of miR-146a. A cell viability analysis was conducted using MTT. A luciferase report vector was used to identify potential targets for miR-146a. Results The miR-146a component was found to be downregulated in glioma tissue compared with adjacent nontumor tissue (p<0.05). The upregulation of miR-146a in glioma cells through miR-146a mimic transfection led to reduction of cell viability and to an increase in the percentage of apoptosis. Notch1 was the name of the potential targeted gene for miR-146a in glioma. Conclusions The study found that the presence of miR-146a potentially affected the proliferation of glioma cells by regulating the rate of Notch1 expression.

scholarly journals Quantification of mRNA in Whole Blood by Assessing Recovery of RNA and Efficiency of cDNA Synthesis

2006 ◽  
Vol 52 (4) ◽  
pp. 634-642 ◽  
Author(s):  
Masato Mitsuhashi ◽  
Shigeru Tomozawa ◽  
Katsuya Endo ◽  
Atsushi Shinagawa
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Whole Blood ◽  
Ex Vivo ◽  
Statistical Significance ◽  
Absolute Quantification ◽  
External Control ◽  
Cdna Synthesis ◽  
Optimized Conditions

Abstract Background: Current gene expression analysis relies on the assumption that the isolated RNA represents all species of mRNA in proportions equal to those in the original materials. No system is available for absolute quantification of mRNA. Methods: We applied whole blood to 96-well filterplates to trap leukocytes. Lysis buffer containing cocktails of specific reverse primers and known concentrations of synthetic external control RNA (RNA34) was added to filterplates, and cell lysates were transferred to oligo(dT)-immobilized microplates for hybridization. We then synthesized the cDNA in the oligo(dT)-immobilized microplates from these primer sites and used the cDNA for real-time PCR. RNA34 acted as a universal control, and gene amplification results were converted to quantities of mRNA per microliter of whole blood after the recovery of RNA34 in each sample was determined. Results: Under fully optimized conditions, both added RNA34 and native mRNA species exhibited ∼10% recovery from whole blood to real-time PCR. When whole blood was stimulated ex vivo, changes in gene expression as low as 30%–40% were detected with statistical significance, and the experimental CVs were low (10%–20%). Conclusion: This new system to estimate mRNA copies per microliter of whole blood may allow standardization of gene-expression–based molecular diagnostics.

Variance Calculations for Quantitative Real-Time PCR Experiments with Multiple Levels of Replication

2013 ◽  
Vol 825 ◽  
pp. 172-176
Author(s):  
Susana Soto-Rojo ◽  
Gary Glonek ◽  
Cecilia Demergasso ◽  
Pedro A. Galleguillos ◽  
Patty Solomon ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Industrial Process ◽  
Error Variance ◽  
Low Grade ◽  
Sulphide Ores ◽  
Polymerase Chain ◽  
Frequent Sampling ◽  
The Relationship ◽  
Proportional Reduction

Heap bioleaching is an established technology for recovering copper from low-grade sulphide ores. Recently, genetics-based approaches have been employed to characterize mineral-processing bacteria. In these approaches, data analysis is a key issue. Consequently, it is of fundamental importance to provide adequate mathematical models and statistical tools to draw reliable conclusions. The present work relates to current studies of the consortium of organisms inhabiting the bioleaching heap of the Escondida mine in Northern Chile. These studies aim to describe and understand the relationship between the dynamics of the community and the performance of the industrial process. Here, we consider a series of quantitative real-time polymerase chain reaction (PCR) experiments performed to quantify six different microorganisms at various stages of the bioleaching cycle. Establishing the reliability of the data obtained by real-time PCR requires the estimation of the error variance at several different levels. The results obtained show that the sampling component of the error variance is the dominant source of variability for most microorganisms. An estimate for the proportional reduction in residual standard deviation from the use of extraction and real-time PCR triplicates was found to range from 3% to 27% for the different organisms. This result suggests that triplicate assays would produce only a modest reduction in error variance compared to more frequent sampling from the heap.

scholarly journals Case report of goose haemorrhagic polyomavirus in 4-day-old goslings indicating vertical transmissibility

2011 ◽  
Vol 80 (3) ◽  
pp. 255-257 ◽  
Author(s):  
Attila Farsang ◽  
Sandor Bernath ◽  
Mihaly Dobos-Kovacs
Keyword(s):  
Case Report ◽  
Disease Control ◽  
Real Time ◽  
Incubation Period ◽  
Vertical Transmission ◽  
Real Time Pcr ◽  
Viral Transmission ◽  
Pcr Assay ◽  
Fatal Disease ◽  
First Time

Haemorrhagic nephritis and enteritis of geese (HNEG) is a fatal disease caused by goose haemorrhagic polyomavirus (GHPV). The aim of our study was to investigate a field outbreak of HNEG by pathological methods and real-time PCR assay using light upon extension (LUX PCR) with special regard to the possibility of vertical transmission. This is the first time that presence of GHPV was confirmed in goslings that died within 4 days after hatching showing typical symptoms of HNEG, which indicates vertical transmissibility as the shortest incubation period of HNEG is 6 days. The way of viral transmission is a key issue and thus the disease control measurements and HNEG epizootiology may be revised based on the findings of this study.

scholarly journals Limitations of Using Propidium Monoazide with qPCR to Discriminate between Live and Dead Legionella in Biofilm Samples

2014 ◽  
Vol 7 ◽  
pp. MBI.S17723 ◽  
Author(s):  
Michael J. Taylor ◽  
Richard H. Bentham ◽  
Kirstin E. Ross
Keyword(s):  
Public Health ◽  
Real Time ◽  
Real Time Pcr ◽  
Culture Method ◽  
Propidium Monoazide ◽  
Factors Affecting ◽  
Conventional Culture ◽  
Target Dna ◽  
The Relationship ◽  
Conventional Culture Method

Accurately quantifying Legionella for regulatory purposes to protect public health is essential. Real-time PCR (qPCR) has been proposed as a better method for detecting and enumerating Legionella in samples than conventional culture method. However, since qPCR amplifies any target DNA in the sample, the technique's inability to discriminate between live and dead cells means that counts are generally significantly overestimated. Propidium monoazide (PMA) has been used successfully in qPCR to aid live/dead discrimination. We tested PMA use as a method to count only live Legionella cells in samples collected from a modified chemostat that generates environmentally comparable samples. Counts from PMA-treated samples that were pretreated with either heat or three types of disinfectants (to kill the cells) were highly variable, with the only consistent trend being the relationship between biofilm mass and numbers of Legionella cells. Two possibilities explain this result: 1. PMA treatment worked and the subsequent muted response of Legionella to disinfection treatment is a factor of biofilm/microbiological effects; although this does not account for the relationship between the amount of biofilm sampled and the viable Legionella count as determined by PMA-qPCR; or 2. PMA treatment did not work, and any measured decrease or increase in detectable Legionella is because of other factors affecting the method. This is the most likely explanation for our results, suggesting that higher concentrations of PMA might be needed to compensate for the presence of other compounds in an environmental sample or that lower amounts of biofilm need to be sampled. As PMA becomes increasingly toxic at higher concentrations and is very expensive, augmenting the method to include higher PMA concentrations is both counterproductive and cost prohibitive. Conversely, if smaller volumes of biofilm are used, the reproducibility of the method is reduced. Our results suggest that using PMA is not an appropriate method for discriminating between live and dead cells to enumerate Legionella for regulatory purposes.

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