scholarly journals DNA Amplification Technique for Detection of Bovine Brucellosis

2018 ◽  
Vol 28 (2) ◽  
pp. 81
Author(s):  
Susan Maphilindawati Noor
Keyword(s):  
Dna Amplification ◽  
Economic Loss ◽  
Molecular Techniques ◽  
Detection Methods ◽  
Bovine Brucellosis ◽  
Cattle Diseases ◽  
Finger Printing ◽  
Amplification Technique ◽  
Polymerase Chain ◽  
Significant Economic Loss

Brucellosis is one of cattle diseases which causes a very significant economic loss and categorized as zoonotic disease. Early detection of Brucellosis in livestock is very important to prevent the spread of disease to livestock and humans. The success of Brucellosis control depends on rapid, sensitive and specific detection methods. The aim of this paper is to review several methods of Brucellosis detection in cattle. Currently, the detection of Brucellosis in Indonesia is using serological and isolation methods. The latter method is the gold standard of Brucellosis diagnosis, however, its sensitivity is low. Therefore, molecular techniques with DNA amplification have been developed and applied in many countries both in livestock and humans because they are more sensitive, specific and rapid in detecting Brucella sp in blood, milk and semen samples. Various DNA amplification methods for detection of Brucellosis that have been developed including polymerase chain reaction (PCR), finger printing and loop-mediated isothermal amplificatiom (LAMP). Both PCR and LAMP are more sensitive and specific in detecting Brucella sp than conventional techniques. PCR technique has advantages in detecting Brucella sp species to serotype and biovar levels. In addition, PCR reagents are cheaper and easier to obtain than LAMP eventhough, LAMP procedure is simpler and faster.

scholarly journals An update on the detection methods of Parachlamydia acanthamoebae, an atypical agent of pneumonia

2019 ◽  
pp. 86-100
Author(s):  
Avinash Rames
Keyword(s):  
Cell Culture ◽  
In Situ Hybridization ◽  
Molecular Techniques ◽  
Detection Methods ◽  
Pathogenic Role ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Polymerase Chain ◽  
Koch Postulates

Parachlamydia acanthamoebae (P. acanthamoebae) has been recognized as an emerging agent of pneumonia as it has been identified in human samples via culture-based, molecular and serological techniques. Additionally, studies on animal models have shown that it fulfills the third and fourth Koch postulates to be assigned a pathogenic role. Due to the threat posed by it, multiple tools have been employed in the search for P. acanthamoebae. The methods utilized for its detection would be cell culture based approaches which involve both animal and amoebal cell culture and also molecular techniques that encompasses polymerase chain reaction (PCR), fluorescence in situ hybridization (FISH) and in situ hybridization (ISH). Additionally, immunohistochemistry (IHC) and serology based techniques such as direct and indirect immunofluorescence are also employed with the usage of Western blotting or immunoblotting as confirmatory procedures. This review attempts to describe the variety of techniques that are present in literature for the isolation and identification of P. acanthamoebae.

Detection ofTrypanosoma congolenseandTrypanosoma bruceisubspecies by DNA amplification using the polymerase chain reaction

Parasitology ◽  
1989 ◽  
Vol 99 (1) ◽  
pp. 57-66 ◽  
Author(s):  
D. R. Moser ◽  
G. A. Cook ◽  
Diane E. Ochs ◽  
Cheryl P. Bailey ◽  
Melissa R. McKane ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Large Scale ◽  
Nuclear Dna ◽  
Pcr Amplification ◽  
Dna Amplification ◽  
Detection Methods ◽  
Chain Reaction ◽  
African Trypanosomes ◽  
Polymerase Chain

SUMMARYThe nuclear DNA ofTrypanosoma congolensecontains a family of highly conserved 369 base pair (bp) repeats. The sequences of three cloned copies of these repeats were determined. An unrelated family of 177 bp repeats has previously been shown to occur in the nuclear DNA ofTrypanosoma brucei brucei(Sloofet al.1983a). Oligonucleotides were synthesized which prime the specific amplification of each of these repetitive DNAs by the polymerase chain reaction (PCR). Amplification of 10% of the DNA in a single parasite ofT. congolenseorT. bruceispp. produced sufficient amplified product to be visible as a band in an agarose gel stained with ethidium bromide. This level of detection, which does not depend on the use of radioactivity, is about 100 times more sensitive than previous detection methods based on radioactive DNA probes. The oligonucleotides did not prime the amplification of DNA sequences in other trypanosome species nor inLeishmania, mouse or human DNAs. Amplification of DNA from the blood of animals infected withT. congolenseand/orT. bruceispp. permitted the identification of parasite levels far below that detectable by microscopic inspection. Since PCR amplification can be conducted on a large number of samples simultaneously, it is ideally suited for large-scale studies on the prevalence of African trypanosomes in both mammalian blood and insect vectors.

scholarly journals Élelmiszer-biztonsági jelentőségű baktériumok molekuláris detektálásának fejlesztése miniatürizált mikrofluidikai eszközök segítségével

2015 ◽  
Vol 156 (51) ◽  
pp. 2082-2088
Author(s):  
Kristóf Iván ◽  
Anna Maráz
Keyword(s):  
Polymerase Chain Reaction ◽  
Pathogenic Bacteria ◽  
Lab On A Chip ◽  
Dna Amplification ◽  
Molecular Techniques ◽  
Microbiological Analysis ◽  
Chain Reaction ◽  
Detection And Identification ◽  
Food Borne ◽  
Polymerase Chain

Detection and identification of food-borne pathogenic bacteria are key points for the assurance of microbiological food safety. Traditional culture-based methods are more and more replaced by or supplemented with nucleic acid based molecular techniques, targeting specific (preferably virulence) genes in the genomes. Internationally validated DNA amplification – most frequently real-time polymerase chain reaction – methods are applied by the food microbiological testing laboratories for routine analysis, which will result not only in shortening the time for results but they also improve the performance characteristics (e.g. sensitivity, specificity) of the methods. Beside numerous advantages of the polymerase chain reaction based techniques for routine microbiological analysis certain drawbacks have to be mentioned, such as the high cost of the equipment and reagents, as well as the risk of contamination of the laboratory environment by the polymerase chain reaction amplicons, which require construction of an isolated laboratory system. Lab-on-a-chip systems can integrate most of these laboratory processes within a miniaturized device that delivers the same specificity and reliability as the standard protocols. The benefits of miniaturized devices are: simple – often automated – use, small overall size, portability, sterility due to single use possibility. These miniaturized rapid diagnostic tests are being researched and developed at the best research centers around the globe implementing various sample preparation and molecular DNA amplification methods on-chip. In parallel, the aim of the authors’ research is to develop microfluidic Lab-on-a-chip devices for the detection and identification of food-borne pathogenic bacteria. Orv. Hetil., 2015, 156(51), 2082–2088.

scholarly journals Advances in nucleic acid-based detection methods.

1992 ◽  
Vol 5 (4) ◽  
pp. 370-386 ◽  
Author(s):  
M J Wolcott
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Nucleic Acid Amplification ◽  
Detection Methods ◽  
Background Information ◽  
Clinical Settings ◽  
Chain Reaction ◽  
Detection Techniques ◽  
Amplification Technique ◽  
Polymerase Chain

Laboratory techniques based on nucleic acid methods have increased in popularity over the last decade with clinical microbiologists and other laboratory scientists who are concerned with the diagnosis of infectious agents. This increase in popularity is a result primarily of advances made in nucleic acid amplification and detection techniques. Polymerase chain reaction, the original nucleic acid amplification technique, changed the way many people viewed and used nucleic acid techniques in clinical settings. After the potential of polymerase chain reaction became apparent, other methods of nucleic acid amplification and detection were developed. These alternative nucleic acid amplification methods may become serious contenders for application to routine laboratory analyses. This review presents some background information on nucleic acid analyses that might be used in clinical and anatomical laboratories and describes some recent advances in the amplification and detection of nucleic acids.

scholarly journals POLYMORPHISM OF SIMPLE SEQUENCE REPEAT REGIONS OF SULAWESI EBONY (DIOSPHYROS CELEBICA BAKH.) IN EXPERIMENTAL FOREST OF HASANUDDIN UNIVERSITY PROVENANCE

2016 ◽  
Vol 1 (1) ◽  
pp. 37-44 ◽  
Author(s):  
Siti Halimah Larekeng ◽  
Muh. Restu ◽  
Gusmiaty Gusmiaty ◽  
Rismawati Rismawati
Keyword(s):  
Dna Isolation ◽  
Pollen Dispersal ◽  
Dna Amplification ◽  
Molecular Techniques ◽  
Dispersal Pattern ◽  
Future Studies ◽  
Annealing Temperatures ◽  
Polymerase Chain ◽  
Simple Sequence ◽  
Dna Isolation Protocol

Polymerase Chain Reaction (PCR)-based molecular techniques have been used to detect the polymorphism in plants. The utilization of molecular markers plays essential role in germplasm characterization and plant breeding since the information of DNA marker technology can be exchanged between laboratories and should have standard method to be reproducible. The molecular aspect has been commonly linked to DNA isolation protocol and polymorphic molecular marker, thus can be used for molecular research recommendation purposes. The objectives of this study were to evaluate the capability of microsatellite marker of Ebenaceae Family for amplifying Ebony DNA, and to determine the appropriate PCR annealing temperatures. The DNA isolation of Ebony leaves from Experimental Forest of Hasanuddin University Provenance was carried out using Genomic DNA Mini Kit (Plant) Geneaid protocol. Nine of seventeen selected primers from the Genus Diospyros were able to amplify Ebony DNA. Amplification products produced polymorphic bands with different annealing temperatures (ranged from 53 to 56°C). These nine polymorphic primers will be recommended to use for future studies in genetic diversity as well as pollen dispersal pattern analyses.

scholarly journals Occurrence of tick-borne diseases in domestic dogs in Belém, Pará, Brazil

2021 ◽  
Vol 15 (4) ◽  
pp. 323-329
Author(s):  
Mylenna de Cássia Neves Guimarães ◽  
Pâmela Talita de Aguiar e Silva ◽  
Thamillys Rayssa Marques Monteiro ◽  
Camila de Cássia dos Santos ◽  
Jacqueline Corrêa Costa ◽  
...  
Keyword(s):  
Hematological Parameters ◽  
Infectious Agent ◽  
Dna Amplification ◽  
Molecular Techniques ◽  
Infectious Agents ◽  
Test Results ◽  
Appropriate Treatment ◽  
Hematological Analysis ◽  
Polymerase Chain ◽  
Veterinary Hospital

Tick-borne blood cell pathogens, which are challenging to diagnose, are primarily detected using molecular techniques. Therefore, this study aimed to detect the main infectious agents involved in 50 cases of suspected hemoparasitosis in dogs treated at the Veterinary Hospital Mário Dias Teixeira of the Federal Rural University of the Amazon. Hematological parameters were evaluated, and blood samples were subjected to polymerase chain reaction (PCR) assays for DNA amplification of the following species: Ehrlichia canis, Anaplasma platys, and Babesia canis. The PCR test results indicated that the most prevalent infectious agent was E. canis, present in 12% (6/50) infected animals, followed by A. platys and B. canis, present in 8% (4/50) and 2% (1/50) infected animals, respectively. Regarding hematological analysis, the most relevant changes were anemia, lymphopenia, thrombocytopenia, leukocytosis, and leukopenia. The availability of molecular techniques allows the management of the most appropriate treatment to infected animals in a rapid and specific way.

scholarly journals Assessment of the Nutraceutical Effects of Oleuropein and the Cytotoxic Effects of Adriamycin, When Administered Alone and in Combination, in MG-63 Human Osteosarcoma Cells

Nutrients ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 354
Author(s):  
Katerina Gioti ◽  
Anastasia Papachristodoulou ◽  
Dimitra Benaki ◽  
Nektarios Aligiannis ◽  
Alexios-Leandros Skaltsounis ◽  
...  
Keyword(s):  
Chemotherapeutic Agent ◽  
Molecular Techniques ◽  
Osteosarcoma Cells ◽  
Elisa Method ◽  
Chain Reaction ◽  
Cytotoxic Effects ◽  
Human Osteosarcoma ◽  
Human Osteosarcoma Cells ◽  
Anti Cancer ◽  
Polymerase Chain

Oleuropein (OLEU) is the most distinguished phenolic compound found in olive fruit and the leaves of Olea europaea L., with several pharmacological properties, including anti-cancer actions. Adriamycin (ADR) is an anthracycline widely used as a chemotherapeutic agent, although it presents significant side effects. The aim of the present study was to investigate the effect of oleuropein alone (20 μg/mL) and in co-treatment with ADR (50 nM), in MG-63 human osteosarcoma cells. Therefore, cellular and molecular techniques, such as MTT assay, flow cytometry, real-time Polymerase Chain Reaction (PCR), western blot and Elisa method, as well as Nuclear Magnetic Resonance (NMR) spectroscopy, were applied to unveil changes in the signal transduction pathways involved in osteosarcoma cells survival. The observed alterations in gene, protein and metabolite levels denote that OLEU not only inhibits MG-63 cells proliferation and potentiates ADR’s cytotoxicity, but also exerts its action, at least in part, through the induction of autophagy.

scholarly journals Impact of Nisin and Nisin-Producing Lactococcus lactis ssp. lactis on Clostridium tyrobutyricum and Bacterial Ecosystem of Cheese Matrices

Foods ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 898
Author(s):  
Hebatoallah Hassan ◽  
Daniel St-Gelais ◽  
Ahmed Gomaa ◽  
Ismail Fliss
Keyword(s):  
Lactococcus Lactis ◽  
Cheddar Cheese ◽  
Economic Loss ◽  
Gas Production ◽  
Clostridium Tyrobutyricum ◽  
Nisin Production ◽  
Genus Clostridium ◽  
Milk Pasteurization ◽  
Cheese Slurry ◽  
Significant Economic Loss

Clostridium tyrobutyricum spores survive milk pasteurization and cause late blowing of cheeses and significant economic loss. The effectiveness of nisin-producing Lactococcus lactis ssp. lactis 32 as a protective strain for control the C. tyrobutyricum growth in Cheddar cheese slurry was compared to that of encapsulated nisin-A. The encapsulated nisin was more effective, with 1.0 log10 reductions of viable spores after one week at 30 °C and 4 °C. Spores were not detected for three weeks at 4 °C in cheese slurry made with 1.3% salt, or during week 2 with 2% salt. Gas production was observed after one week at 30 °C only in the control slurry made with 1.3% salt. In slurry made with the protective strain, the reduction in C. tyrobutyricum count was 0.6 log10 in the second week at 4 °C with both salt concentration. At 4 °C, nisin production started in week 2 and reached 97 µg/g after four weeks. Metabarcoding analysis targeting the sequencing of 16S rRNA revealed that the genus Lactococcus dominated for four weeks at 4 °C. In cheese slurry made with 2% salt, the relative abundance of the genus Clostridium decreased significantly in the presence of nisin or the protective strain. The results indicated that both strategies are able to control the growth of Clostridium development in Cheddar cheese slurries.

scholarly journals Rapid Multi-Hybridisation FISH Screening for Balanced Porcine Reciprocal Translocations Suggests a Much Higher Abnormality Rate Than Previously Appreciated

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 250
Author(s):  
Rebecca E O’Connor ◽  
Lucas G Kiazim ◽  
Claudia C Rathje ◽  
Rebecca L Jennings ◽  
Darren K Griffin
Keyword(s):  
Semen Analysis ◽  
In Situ Hybridisation ◽  
Economic Loss ◽  
Environmental Damage ◽  
Fluorescence In Situ Hybridisation ◽  
Reciprocal Translocations ◽  
Chromosome Translocations ◽  
Farrowing Rate ◽  
Significant Economic Loss

With demand rising, pigs are the world’s leading source of meat protein; however significant economic loss and environmental damage can be incurred if boars used for artificial insemination (AI) are hypoprolific (sub-fertile). Growing evidence suggests that semen analysis is an unreliable tool for diagnosing hypoprolificacy, with litter size and farrowing rate being more applicable. Once such data are available, however, any affected boar will have been in service for some time, with significant financial and environmental losses incurred. Reciprocal translocations (RTs) are the leading cause of porcine hypoprolificacy, reportedly present in 0.47% of AI boars. Traditional standard karyotyping, however, relies on animal specific expertise and does not detect more subtle (cryptic) translocations. Previously, we reported development of a multiple hybridisation fluorescence in situ hybridisation (FISH) strategy; here, we report on its use in 1641 AI boars. A total of 15 different RTs were identified in 69 boars, with four further animals XX/XY chimeric. Therefore, 4.5% had a chromosome abnormality (4.2% with an RT), a 0.88% incidence. Revisiting cases with both karyotype and FISH information, we reanalysed captured images, asking whether the translocation was detectable by karyotyping alone. The results suggest that chromosome translocations in boars may be significantly under-reported, thereby highlighting the need for pre-emptive screening by this method before a boar enters a breeding programme.

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