Molecular detection of Brucella in milk using polymerase chain reaction

Brucellosis is a highly contagious disease affecting a wide variety of farm animals. It is also an important zoonosis, and man is often infected following contact with infected animals or the consumption of contaminated milk and milk products. At present, mainly bacteriological and serological detection methods are used. A bacteriological method takes days to weeks to grow the organism besides its health hazard. Serological tests are faster but antigen–antibody interactions can be faulted by non-specific interactions. A method for direct detection of Brucella melitensis in 1 ml of milk was developed on the basis of enzymatic treatment of milk components and subsequent PCR and line probe assay (LPA). After PCR, 3 × 104 CFU/ml sensitivity was obtained by agarose gel electrophoresis and LPA. The safety and sensitivity of LPA combined with its speed suggests the potential of this technique for diagnosis of brucellosis in milk rather than the time consuming classical methods.

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Canine ehrlichiosis: clinical, hematological, serological and molecular aspects

Ciência Rural ◽

10.1590/s0103-84782008000300027 ◽

2008 ◽

Vol 38 (3) ◽

pp. 766-770 ◽

Cited By ~ 36

Author(s):

Andréa Cristina Higa Nakaghi ◽

Rosangela Zacarias Machado ◽

Mirela Tinucci Costa ◽

Marcos Rogério André ◽

Cristiane Divan Baldani

Keyword(s):

Direct Detection ◽

Clinical Signs ◽

Chronic Phase ◽

Antibody Test ◽

Acute Stage ◽

Detection Methods ◽

Serological Tests ◽

Humoral Immune ◽

Canine Ehrlichiosis ◽

Dot Elisa

The aim of the present study was to compare the direct detection methods of Ehrlichia canis (blood smears and nested PCR), serological tests (Dot-ELISA and Immunofluorescent Antibody Test - IFAT), and demonstrate the most suitable test for the diagnosis of different stages of infection. Blood samples and clinical data were collected from 30 dogs examined at the Veterinary Teaching Hospital, UNESP, Jaboticabal, SP, Brazil. The clinical signs most frequently observed were apathy, anorexia, pale mucous membrane, fever, lymphadenopathy, splenomegaly, hemorrhages and uveitis. Evaluating the humoral immune response, 63.3% of the sera were IFAT positive, while 70% were Dot-ELISA positive. By nestedPCR 53.3% of the samples were positive. Comparing these techniques it was concluded that serology and nPCR are the most suitable tests to confirm the diagnosis of canine ehrlichiosis, however it should be always treated as a complementary data to clinical and hematological evaluation. Serology has an important role in the subclinical and in the chronic phase, nPCR is recommended in the acute stage, and, especially, to identify the ehrlichia specie.

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In VitroDiagnostic Device with a Bio-Coupled Gate Field Effect Transistor

International Journal of Automation Technology ◽

10.20965/ijat.2018.p0045 ◽

2018 ◽

Vol 12 (1) ◽

pp. 45-51 ◽

Cited By ~ 1

Author(s):

Toshiya Sakata ◽

Keyword(s):

Semiconductor Device ◽

Direct Detection ◽

Cell Communication ◽

Label Free ◽

Cellular Functions ◽

Biological Phenomena ◽

Communication Processes ◽

Antigen Antibody ◽

Antibody Interactions

In this study, we report a simple and rapid biosensing method for the analysis andin vitromonitoring of biological processes, including DNAbinding events, antigen-antibody interactions, and cellular functions, using a semiconductor device. Most biological phenomena involve cell-cell communication processes that are mediated by the transport of sodium or potassium ions and other charged biomolecules, such as DNA, across ion channels in the cell membrane. Therefore, our approach focused on the direct detection of changes in ion concentrations by utilizing a semiconductor-based biosensor device. Our results demonstrated that our semiconductor-based biosensor platform achieves label-free and noninvasive biosensing that is suitable forin vitrodiagnosis.

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Investigation on Brucella infection in farm animals in Saham, Sultanate of Oman with reference to human brucellosis outbreak

BMC Veterinary Research ◽

10.1186/s12917-019-2093-4 ◽

2019 ◽

Vol 15 (1) ◽

Cited By ~ 1

Author(s):

Yasmin ElTahir ◽

Anfal Al-Farsi ◽

Waleed Al-Marzooqi ◽

Alghalya Al-Toobi ◽

Osman M. Gaafar ◽

...

Keyword(s):

Brucella Melitensis ◽

Phase 1 ◽

Time Frame ◽

Farm Animals ◽

Human Brucellosis ◽

Biochemical Tests ◽

Serological Tests ◽

Molecular Tests ◽

The Status ◽

Bronchial Lymph Node

Abstract Background The objective of this study was to investigate Brucella infection in farm animals in Saham, Oman, with reference to a survey carried out by the Ministry of Agriculture & Fisheries (MAF) for Brucellosis during the period of May to July 2016 in Saham, following an outbreak of human brucellosis. We wanted to apply different serological, bacteriological and molecular tests in a time frame (phase 1, 2 & 3) with reference to the pivotal time of a human brucellosis outbreak to ascertain the status of the disease in Saham area where the MAF survey was conducted. Blood samples were collected from farm animals and sera were screened in parallel for Brucella antibodies using different serological tests. Results Using the RBT test, phase 1 sera showed seropositivity in sheep at 2.6%, (95% CI: 0.5–13.5%), in camel (5.9%, 1.1–27.0%), but not in sera from goats and cattle (0%). Using I-ELISA, seropositivity in goat was 3.1% (0.6–15.8%), with no positive sheep and cattle. Using c-ELISA for camel we found a seropositivity of 5.9% (1.1–27.0%). Furthermore, CFT seropositivity in goats was 21.9% (CI: 11.3–38.9), cattle and sheep sera were negative and camel was 5.9% (1.1–27.0%). In phase 2, the seropositivity in goats was 1.9% (1.4–2.6%), sheep 4.5% (3.5–5.8%), cattle 1.1%, (0.5–2.3%) and camels 18.2% (5.1–47.7%), Phase 3 sera were collected 6 months after the human brucellosis outbreak. With RBT, the seropositivity in goats was 3% (1.0–8.5%), sheep 2% (0.6–7.1%) cattle 1% (0.2–5.5%). With I-ELISA, goats & camels were negative, sheep were 3% (1.0–8.5%) and cattle 1% (0.2–5.5%). Moreover, B. melitensis was isolated from a bronchial lymph node of the RBT and I-ELISA seropositive cow and confirmed by Multiplex PCR and biochemical tests. Conclusion Using a retrospective study analysis of animal sera and following up after a human brucellosis outbreak, the present study showed a slight decrease in seropositivity of infected animals after the MAF implemented test and slaughter policy. The most interesting finding in this study was the isolation, identification and molecular characterization of Brucella melitensis in a cow (spillover), which is not a preferential host for Brucella melitensis.

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Viruses and Water: Problems, Detection, and Control

Water Science & Technology ◽

10.2166/wst.1993.0543 ◽

1993 ◽

Vol 27 (7-8) ◽

pp. 127-133 ◽

Cited By ~ 2

Author(s):

H. Dizer ◽

J. Dürkop ◽

A. Grohmann ◽

H. Kopecka ◽

J. M. López-Pila

Keyword(s):

Drinking Water ◽

Surface Waters ◽

Membrane Filtration ◽

Wastewater Treatment Plants ◽

Health Hazard ◽

Primary Source ◽

Detection Methods ◽

Secondary Effluent ◽

Naturally Occurring ◽

Water Problems

Secondary effluent of wastewater treatment plants contains a high number of viruses and other pathogens, which pose a health risk to the population, (especially when receiv ng waters are used for bathing and swimming, or for growing shellfish. In areas with a high density of population, where drinking water supply is dependent on surface waters and contaminated rivers are the primary source of drinking water, failure of the filtration or of the disinfection step, or of any other “barriers” supposed to warrant safe potable water, will increase the risk of health hazard for the consumer. We have compared the efficiency of viral elimination in secondary effluent by flocculation, uv rradiation and membrane filtration taking naturally occurring, or additionally seeded f2 phages, as indicator for viruses. Flocculation decreased the number of phages present in secondary effluent by more than two logs. If combined with uv irradiation, the elimination reached five additional logs. Membrane filtration eliminated essentially all naturally occurring phages. Improvement of the quality of surface waters calls for a refinement of detection methods for viruses. We have found that the polymerase chain reaction (PCR) might be used for detecting viruses in surface waters.

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A brief review of diagnosis of small ruminants brucellosis

Current Medicinal Chemistry ◽

10.2174/0929867328666201231121226 ◽

2020 ◽

Vol 28 ◽

Author(s):

Jingjing Ren ◽

Qisheng Peng

Keyword(s):

Public Health ◽

Infectious Disease ◽

Economic Impact ◽

False Positive ◽

Brucella Melitensis ◽

Small Ruminants ◽

Animal Husbandry ◽

Detection Methods ◽

Immunological Tests

: Brucellosis caused by bacteria of the genus of Brucella remains a major zoonosis in the widely world, which is an infectious disease with a severe economic impact on animal husbandry and public health. The genus of Brucella includes ten species and the most prevalent is Brucella melitensis. The diagnosis of Brucella melitensis ruminant brucellosis is based on bacteriological and immunological tests. The use of vaccines and the false-positive serological reactions (FPSR) caused by other cross-reacting bacteria represent the immunological contexts. This complex context results in the development of the large number of diagnosis of Brucella melitensis brucellosis. The aim of this article is to briefly review the detection methods and compare the superiorities of different tests.

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Affinity Sensors for the Diagnosis of COVID-19

Micromachines ◽

10.3390/mi12040390 ◽

2021 ◽

Vol 12 (4) ◽

pp. 390

Author(s):

Maryia Drobysh ◽

Almira Ramanaviciene ◽

Roman Viter ◽

Arunas Ramanavicius

Keyword(s):

Field Effect Transistors ◽

Resonance Field ◽

Analytical Signal ◽

Nucleic Acid Hybridization ◽

Detection Methods ◽

Label Free ◽

Infected People ◽

Label Free Detection ◽

Main Instrument ◽

Antigen Antibody

The coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was proclaimed a global pandemic in March 2020. Reducing the dissemination rate, in particular by tracking the infected people and their contacts, is the main instrument against infection spreading. Therefore, the creation and implementation of fast, reliable and responsive methods suitable for the diagnosis of COVID-19 are required. These needs can be fulfilled using affinity sensors, which differ in applied detection methods and markers that are generating analytical signals. Recently, nucleic acid hybridization, antigen-antibody interaction, and change of reactive oxygen species (ROS) level are mostly used for the generation of analytical signals, which can be accurately measured by electrochemical, optical, surface plasmon resonance, field-effect transistors, and some other methods and transducers. Electrochemical biosensors are the most consistent with the general trend towards, acceleration, and simplification of the bioanalytical process. These biosensors mostly are based on the determination of antigen-antibody interaction and are robust, sensitive, accurate, and sometimes enable label-free detection of an analyte. Along with the specification of biosensors, we also provide a brief overview of generally used testing techniques, and the description of the structure, life cycle and immune host response to SARS-CoV-2, and some deeper details of analytical signal detection principles.

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Contamination of bovine, sheep and goat meat with Brucella spp.

Italian Journal of Food Safety ◽

10.4081/ijfs.2016.5913 ◽

2016 ◽

Vol 5 (3) ◽

Cited By ~ 1

Author(s):

Francesco Casalinuovo ◽

Lucia Ciambrone ◽

Antonio Cacia ◽

Paola Rippa

Keyword(s):

Lymph Nodes ◽

Brucella Abortus ◽

Brucella Melitensis ◽

Chain Reaction ◽

Internal Organs ◽

Goat Meat ◽

Real Risk ◽

Bacteriological Method ◽

Polymerase Chain ◽

Type 3

A study was conducted in order to evaluate the contamination by Brucella spp. of meat from animals slaughtered because they resulted positive for brucellosis at some time during their life, given that European legislation allows these meats to be freely marketed. After slaughter and before delivery to market outlets, swab samples were taken from 307 carcasses of infected animals: 40 cattle, 60 sheep and 207 goats. The swabs were subsequently analyzed by means of polymerase chain reaction (PCR) tests. In addition, bacteriological tests were carried out on the lymph nodes and internal organs of the same animals. Brucella spp. was detected by means of PCR in 25/307 carcasses (8%): 1 bovine (2.5%), 9 sheep (15%) and 15 goats (7.2%) and was isolated, by means of a cultural method, in 136/307 carcasses (44%). Moreover, additional analysis, performed on lymph nodes from the same carcasses that had proved positive by PCR allowed to highlight type 3 Brucella abortus in the bovine carcass and type 3 Brucella melitensis in the sheep and goat carcasses. The study shows that the cattle, sheep and goats meat of animals slaughtered because they have tested positive for brucellosis, may be contaminated by Brucella spp. As this could constitute a real risk of transmission to both butchery personnel and consumers, so the meat of animals infected by Brucella spp. should be analyzed before being marketed and, PCR technique performed on swabs proved more useful, practical and faster than the traditional bacteriological method.

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Biosensors for the Determination of SARS-CoV-2 Virus and Diagnosis of COVID-19 Infection

International Journal of Molecular Sciences ◽

10.3390/ijms23020666 ◽

2022 ◽

Vol 23 (2) ◽

pp. 666

Author(s):

Maryia Drobysh ◽

Almira Ramanaviciene ◽

Roman Viter ◽

Chien-Fu Chen ◽

Urte Samukaite-Bubniene ◽

...

Keyword(s):

Current Trend ◽

Analytical Signal ◽

Diagnostic Methods ◽

Detection Methods ◽

Label Free ◽

Free Mode ◽

Rna Hybridisation ◽

Antigen Antibody ◽

Affinity Interaction

Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity.

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Seroprevalance of Toxoplasmosis in sheep and goat: Iraq/ Sulaimania

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v35i1.599 ◽

2011 ◽

Vol 35 (1) ◽

pp. 16-24

Author(s):

Lazem H. Al-Taie

Keyword(s):

Age Groups ◽

Neural System ◽

Farm Animals ◽

Economic Losses ◽

Age Group ◽

Serum Samples ◽

Serological Tests ◽

Back Ground ◽

Qualitative Determination

Back ground: Toxoplasmosis is an important zoonosis that causes economic losses in animal herds due to abortion and stillbirth as well as changes in the reproductive and neural system of susceptible animals . Objective: The aims of the present study is to determination the prevalence of T. gondii in farm animals ( sheep& goat)of both genders and different ages in Sulaimani province by using two serological tests (ELISA and LAT). Methods: Blood samples were collected from farm animals ,142 sheep and 46 goats , of different sexes and ages. Tow different serological tests ,ELISA and LAT for qualitative determination of T. gondii antibody titer in sheep and goats serum samples. Results: The prevalence rate in sheep was 73 (51.7 %) and 82 (57 %) , and 21 (54.6 %) and 25 (54.35 %) in goats ,by ELISA and LAT respectively. The prevalence of toxoplasmosis was highest in age group 7-9 (66.6%) in sheep in compares’ with other age groups. There was no significant differences between both spp.and tow test. Conclusion: Statistical results show no significant differences between both tests (ELISA &LAT) at (P ≥ 0.05).The prevalence of toxoplasmosis was increased proportionally with the age of animals, while gender has no effect on the prevalent rate .

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