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

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.

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Molecular diagnosis of Ehrlichia canis infection in dogs with uveitis

Semina Ciências Agrárias ◽

10.5433/1679-0359.2018v39n3p1049 ◽

2018 ◽

Vol 39 (3) ◽

pp. 1049

Author(s):

Jéssica Fontes Veloso ◽

Leonardo Sauer ◽

Arianne Pontes Oriá ◽

Deusdete Conceição Gomes Junior ◽

Ana Cláudia Santos Raposo ◽

...

Keyword(s):

Molecular Diagnosis ◽

Infectious Agent ◽

Posterior Uveitis ◽

Ehrlichia Canis ◽

State University ◽

Nested Polymerase Chain Reaction ◽

Canine Monocytic Ehrlichiosis ◽

Polymerase Chain ◽

Clinical Changes ◽

Veterinary Hospital

Canine monocytic ehrlichiosis (CME) is an infectious disease caused by a gram-negative bacterium Ehrlichia canis that has a high global prevalence that leads to high rates of morbidity and mortality in dogs. Among the clinical changes, ophthalmic diseases can lead to permanent blindness and it can be an important clinical sign. The objective of this study was to perform nested polymerase chain reaction (PCR) to diagnose E. canis infection in dogs with bilateral uveitis from the Veterinary Hospital of the Santa Cruz State University. Blood samples were collected and DNA for the molecular diagnosis was extracted from 66 adult dogs of both genders and mixed breeds diagnosed with bilateral uveitis. Thirty-five (53%) dogs showed positive results and presented with iridocyclitis, posterior uveitis, panuveitis, or uveitis with secondary glaucoma. This study demonstrates that nested PCR is an important tool for the differential diagnosis of dogs with bilateral uveitis, as it provides evidence of the infectious agent in the animal.

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DNA Amplification Technique for Detection of Bovine Brucellosis

Indonesian Bulletin of Animal and Veterinary Sciences ◽

10.14334/wartazoa.v28i2.1829 ◽

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.

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Real-time isothermal DNA amplification monitoring in picoliter volumes using an optical fiber sensor

Lab on a Chip ◽

10.1039/d0lc01069c ◽

2021 ◽

Author(s):

Monika Janik ◽

Seyed Vahid Hamidi ◽

Marcin Koba ◽

Jonathan Perreault ◽

Ryan Walsh ◽

...

Keyword(s):

Gold Standard ◽

Rolling Circle Amplification ◽

Optical Fiber Sensor ◽

Dna Amplification ◽

Fiber Sensor ◽

Test Results ◽

Chain Reaction ◽

Rolling Circle ◽

Standard Polymerase Chain Reaction ◽

Polymerase Chain

Rolling circle amplification (RCA) of DNA can be considered as a great alternative to the gold standard polymerase chain reaction (PCR), where rapid, sensitive, and reliable test results are required.

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

Orvosi Hetilap ◽

10.1556/650.2015.30325 ◽

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.

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Estimation of Hematological Parameters of Disease Severity in Iraqi Patients with COVID-19

Iraqi Journal of Science ◽

10.24996/ijs.2021.62.10.8 ◽

2021 ◽

pp. 3487-3496

Author(s):

Zainab S. Mahmood ◽

Hula Y. Fadhil ◽

Ali H Ad′hiah

Keyword(s):

Hematological Parameters ◽

Systemic Disease ◽

Severe Disease ◽

Local Data ◽

Substantial Impact ◽

Main Indicator ◽

Hematological Analysis ◽

Polymerase Chain ◽

Blood Specimens ◽

Severity Of The Disease

Coronavirus disease 2019 (COVID-19) is a systemic disease with a substantial impact on the hematopoietic system and hemostasis. Neutrophilia is an early indicator of SARS-CoV-2 infection, while lymphopenia acts as a biomarker of the severity of infection, and the neutrophil-to-lymphocyte ratio (NLR) is the main indicator of cytokine storms. Thus, this study aimed to provide local data about hematological parameters among COVID-19 patients and estimate their correlation with viral load and other factors in severe cases. A total of 99 nasopharyngeal swabs and whole blood specimens were collected from individuals suspected with COVID-19 between October and December 2020. Samples were tested by real time reverse transcriptase polymerase chain reaction (rRT-PCR) assay, COVID-19 IgG and IgM antibody tests, beside hematological analysis. The results showed a significant increase in neutrophils count and NLR, correlated with the severity of the disease Patients at older ages who are suffering from some comorbidity like hypertension and diabetes are at elevated risk to develop more severe disease outcome. The findings demonstrated a higher neutrophils count and higher death ratio in older ages. Also, the results suggest that NLR could be employed as a useful biomarker and potential prognostic tool supporting the importance of laboratory records in assessing case severity and disease progression.

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

AgroTech Journal ◽

10.31327/atj.v1i1.173 ◽

2016 ◽

Vol 1 (1) ◽

pp. 37-44 ◽

Cited By ~ 4

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.

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Update on molecular diagnosis of human leptospirosis

Asian Biomedicine ◽

10.1515/abm-2019-0063 ◽

2020 ◽

Vol 13 (6) ◽

pp. 207-216

Author(s):

Teerasit Techawiwattanaboon ◽

Kanitha Patarakul

Keyword(s):

Molecular Detection ◽

Molecular Techniques ◽

Morbidity And Mortality ◽

Chain Reaction ◽

Clinical Specimens ◽

Human Leptospirosis ◽

Molecular Approaches ◽

Resource Limited ◽

Appropriate Treatment ◽

Polymerase Chain

AbstractBackgroundLeptospirosis, caused by pathogenic Leptospira spp., is a widespread zoonotic disease worldwide. Early diagnosis is required for proper patient management and reducing leptospirosis morbidity and mortality.ObjectiveTo summarize current literature regarding commonly used and new promising molecular approaches to Leptospira detection and diagnostic tests of human leptospirosis.MethodThe relevant articles in Leptospira and leptospirosis were retrieved from MEDLINE (PubMed) and Scopus.ResultsSeveral molecular techniques have been developed for diagnosis of human leptospirosis. Polymerase chain reaction-based techniques targeting on either lipL32 or 16S rRNA (rrs) gene are most commonly used to detect leptospiral DNA in various clinical specimens. Whole blood and urine are recommended specimens for suspected cases in the first (acute) and the second (immune) phases, respectively. Isothermal amplification with less expensive instrument is an alternative DNA detection technique that may be suitable for resource-limited laboratories.ConclusionDetection of leptospiral DNA in clinical specimens using molecular techniques enhances sensitivity for diagnosis of leptospirosis. The efficient and robust molecular detection especially in the early leptospiremic phase may prompt early and appropriate treatment leading to reduced morbidity and mortality of patients with leptospirosis.

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Exposure of the Gulf of St. Lawrence grey seal Halichoerus grypus population to potentially zoonotic infectious agents

Diseases of Aquatic Organisms ◽

10.3354/dao03536 ◽

2020 ◽

Vol 142 ◽

pp. 105-118

Author(s):

CC Sauvé ◽

A Hernández-Ortiz ◽

E Jenkins ◽

F Mavrot ◽

A Schneider ◽

...

Keyword(s):

Meat Products ◽

Halichoerus Grypus ◽

Human Consumption ◽

Infectious Agents ◽

Erysipelothrix Rhusiopathiae ◽

Public Health Significance ◽

Domestic Livestock ◽

Health Significance ◽

Commercial Source ◽

Polymerase Chain

The population of grey seals Halichoerus grypus in Canadian waters is currently used as a commercial source of meat for human consumption. As with domestic livestock, it is important to understand the occurrence in these seals of infectious agents that may be of public health significance and thus ensure appropriate measures are in place to avoid zoonotic transmission. This study examined the prevalence of antibodies against Brucella spp., Erysipelothrix rhusiopathiae, 6 serovars of Leptospira interrogans, and Toxoplasma gondii in 59 grey seals and determined by polymerase chain reaction (PCR) the presence of these potentially zoonotic agents in specific organs and tissues of seropositive animals. The presence of encysted Trichinella spp. larvae was also investigated by digestion of tongue, diaphragm and other muscle samples, but none were detected. Seroprevalence against Brucella spp. and E. rhusiopathiae was low (5 and 3%, respectively). All 59 seals tested had antibodies against L. interrogans, but no carrier of this bacterium was detected by PCR. Seroprevalence against T. gondii was 53%, and DNA of this protozoan was detected by PCR in 11/30 (37%) seropositive animals. Standard sanitary measures mandatory for commercialization of meat products for human consumption should greatly reduce the potential for exposure to these infectious agents. However, special consideration should be given to freezing seal meat for at least 3 d to ensure destruction of tissue cysts of T. gondii.

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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 ◽

10.3390/nu13020354 ◽

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.

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Is it enough for COVID-19 screening test? Limitation of swab test and general characteristics of mild symptom patients

Science Progress ◽

10.1177/00368504211026152 ◽

2021 ◽

Vol 104 (2) ◽

pp. 003685042110261

Author(s):

Sungwoo Choi ◽

Hyo Jeong Choi ◽

Ho Jung Kim

Keyword(s):

Screening Test ◽

Community Treatment ◽

Upper Respiratory Tract ◽

Test Results ◽

Rt Pcr ◽

Pcr Assay ◽

Positive Rate ◽

Upper Respiratory ◽

Polymerase Chain ◽

Discharge Criteria

The most common method for SARS-CoV-2 testing is throat or nasal swabbing by real-time reverse transcription polymerase chain reaction (RT-PCR) assay. In South Korea, drive-through swab test is used for screening system and community treatment centers (CTCs), which admit and treat confirmed COVID-19 patients with mild symptoms, are being used. This retrospective study was conducted on patients admitted to a CTC on March 6, 2020. A total of 313 patients were admitted. The nasal and throat swabs were collected from the upper respiratory tract, and a sputum test was performed to obtain lower respiratory samples. The positive rate of the first set of test, sputum test was higher than that of the swab test ( p = 0.011). In the second set of test, 1 week after the first ones, the rate of positive swab tests was relatively high ( p = 0.026). In the first set of test, 66 of 152 (43.4%) patients showed 24-h consecutive negative swab test results, when the sputum test results were considered together, that number fell to 29 patients (19.1%) ( p < 0.001). Also, in the second set of test, 63 of 164 (38.4%) patients met the discharge criteria only when the swab test was considered; that number fell to 30 (18.3%) when the sputum test results were also considered ( p < 0.001). Using the swab test alone is insufficient for screening test and discharge decision. Patients who may have positive result in the sputum test can be missed.

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