scholarly journals Development of High-Throughput Multiplex Serology to Detect Serum Antibodies against Coxiella burnetii

2021 ◽  
Vol 9 (11) ◽  
pp. 2373
Author(s):  
Rima Jeske ◽  
Larissa Dangel ◽  
Leander Sauerbrey ◽  
Dimitrios Frangoulidis ◽  
Lauren R. Teras ◽  
...  

The causative agent of Q fever, the bacterium Coxiella burnetii (C. burnetii), has gained increasing interest due to outbreak events and reports about it being a potential risk factor for the development of lymphomas. In order to conduct large-scale studies for population monitoring and to investigate possible associations more closely, accurate and cost-effective high-throughput assays are highly desired. To address this need, nine C. burnetii proteins were expressed as recombinant antigens for multiplex serology. This technique enables the quantitative high-throughput detection of antibodies to multiple antigens simultaneously in a single reaction. Based on a reference group of 76 seropositive and 91 seronegative sera, three antigens were able to detect C. burnetii infections. Com1, GroEL, and DnaK achieved specificities of 93%, 69%, and 77% and sensitivities of 64%, 72%, and 47%, respectively. Double positivity to Com1 and GroEL led to a combined specificity of 90% and a sensitivity of 71%. In a subgroup of seropositives with an increased risk for chronic Q fever, the double positivity to these markers reached a specificity of 90% and a sensitivity of 86%. Multiplex serology enables the detection of antibodies against C. burnetii and appears well-suited to investigate associations between C. burnetii infections and the clinical manifestations in large-scale studies.

Author(s):  
Attila Dobos ◽  
István Fodor ◽  
Gerda Kiss ◽  
Miklós Gyuranecz

AbstractQ fever is a disease of high zoonotic potential, but interest in its causative agent is rather low although it causes some public health problems in Hungary. The prevalence of Q fever is highly variable by country. The main reservoirs of the disease are the same domestic ruminant species everywhere, but the epidemiological profile depends on the features of the specific reservoir. The aim of this large-scale study was to demonstrate the importance of Q fever in different species as a possible source for human infection in most regions of Hungary. A total of 851 serum samples from 44 dairy farms, 16 sheep flocks, 4 goat farms and 3 zoos located in different parts of Hungary were tested. The presence of antibodies to Coxiella burnetii was surveyed in dairy cattle (n = 547), goats (n = 71), sheep (n = 200) and zoo animals (n = 33). The animal species tested in Hungary showed different seroprevalence values of C. burnetii infection. Seropositivity by the enzyme-linked immunosorbent assay was found in 258 out of 547 (47.2%) cows and in 69 out of 271 (25.5%) small ruminants, among them in 47 out of 200 (23.5%) sheep and in 22 out of 71 (31.0%) goats. Antibodies to C. burnetii were not detected in zoo animals. Seropositivity was demonstrated in 44 out of 44 (100%) dairy cattle farms, with at least one serum sample found to be positive on each farm. The seropositivity rate of small ruminant farms was 55.0% (11 positive out of 20 tested), with 9 out of 16 (56.3%) sheep flocks and 2 out of 4 (50.0%) goat herds showing seropositivity.


Healthcare ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 96 ◽  
Author(s):  
Courtney Scherr ◽  
Sharon Aufox ◽  
Amy Ross ◽  
Sanjana Ramesh ◽  
Catherine Wicklund ◽  
...  

From a public health perspective, the “All of Us” study provides an opportunity to isolate targeted and cost-effective prevention and early-detection strategies. Identifying motivations for participation in large-scale genomic sequencing (LSGS) studies, and motivations and preferences to receive results will help determine effective strategies for “All of Us” study implementation. This paper offers a critical review of the literature regarding LSGS for adult onset hereditary conditions where results could indicate an increased risk to develop disease. The purpose of this review is to synthesize studies which explored peoples’ motivations for participating in LSGS studies, and their desire to receive different types of genetic results. Participants were primarily motivated by altruism, desire to know more about their health, and curiosity. When asked about hypothetically receiving results, most participants in hypothetical studies wanted all results except those which were uncertain (i.e., a variant of uncertain significance (VUS)). However, participants in studies where results were returned preferred to receive only results for which an intervention was available, but also wanted VUS. Concerns about peoples’ understanding of results and possible psychosocial implications are noted. Most studies examined populations classified as “early adopters,” therefore, additional research on motivations and expectations among the general public, minority, and underserved populations is needed.


2013 ◽  
Vol 35 (2) ◽  
pp. 270-270 ◽  
Author(s):  
Hongzhi Cao ◽  
Yu Wang ◽  
Wei Zhang ◽  
Xianghua Chai ◽  
Xiandong Zhang ◽  
...  

2014 ◽  
Vol 7 (2) ◽  
pp. 153-166 ◽  
Author(s):  
F. Cheli ◽  
E. Fusi ◽  
A. Baldi

This review presents the applications of cell-based models in mycotoxin research, with a focus on models for mycotoxin screening and cytotoxicity evaluation. Various cell-based models, cell and cell culture condition related factors, toxicity endpoints and culture systems as well as predictive value of cell-based bioassays are reviewed. Advantages, drawbacks and technical problems regarding set up and validation of consistent, robust, reproducible and high-throughput cell-based models are discussed. Various cell-based models have been developed and used as screening tests for mycotoxins but the data obtained are difficult to compare. However, the results highlight the potential of cell-based models as promising in vitro platforms for the initial screening and cytotoxicity evaluation of mycotoxins and as a significant analytical approach in mycotoxin research before any animal or human clinical studies. To develop cell-based models as powerful high-throughput laboratory platforms for the analysis of large numbers of samples, there are mainly two fundamental requirements that should be met, i.e. the availability of easy-to-use and, if possible, automated cell platforms and the possibility to obtain reproducible results that are comparable between laboratories. The transition from a research model to a test model still needs optimisation, standardisation, and validation of analytical protocols. The validation of a cell-based bioassay is a complex process, as several critical points, such as the choice of the cellular model, the assay procedures, and the appropriate use and interpretation of the results, must be strictly defined to ensure more consistency in the results. The development of cell-based models exploring the third dimension together with automation and miniaturisation will bring cellular platforms to a level appropriate for cost-effective and large-scale analysis in the field of mycotoxin research.


Blood ◽  
2016 ◽  
Vol 127 (1) ◽  
pp. 113-121 ◽  
Author(s):  
Cléa Melenotte ◽  
Matthieu Million ◽  
Gilles Audoly ◽  
Audrey Gorse ◽  
Hervé Dutronc ◽  
...  

Key PointsCoxiella burnetii is associated with an increased risk of lymphoma; its presence in the tumor microenvironment may favor lymphomagenesis. Lymphoma has to be considered in patients with Q fever and lymphoid disorders, especially those with persistent focalized infections.


2017 ◽  
Vol 20 (4) ◽  
pp. 190-193

Q fever is caused by an anthropozoonosis determined by the pathogen Coxiella burnetii, a gram-negative bacterium with intracellular growth. The occurrence of infection in the human species takes place through inhalation of contaminated aerosols or dust from infected domestic animals (cattle, sheep, goats) and more rarely through ingestion of unpasteurized milk, infected mite or inter-human transmission. The endocardium is one of the main infection sites, especially in the context of the long-term development of the disease, and cardiac decompensation often leads to death in absence of a proper diagnosis and appropriate treatment (1).We present the case of a patient of the male sex aged 37 years without personal pathologic history known admitted in “St. Parascheva” Clinical Hospital for Infectious Diseases Iasi complaining of productive cough, fatigue, shortness of breath with moderate effort and pain in the left scapulohumeral joint with irradiation in the left upper limb. Clinical examination objectified digital clubbing, systolic/diastolic murmurs throughout the precordium area and hepato-splenomegaly, while laboratory tests revealed the presence of inflammatory syndrome, cholestasis and hepatic cytolysis. Echocardiography shows a hyperechogenic entity at the level of the aortic valve, as well as a severe valve disorder. The diagnosis of infective endocarditis is established on aortic valve and therapy with first-choice antibiotics, consisting of triple combination of cefotaxime, amikacin and vancomycin, is initiated. Blood cultures taken upon admission were negative, while positive serological phase I and II tests for C. burnetii urged the indication of changing therapy with doxycycline and trimethoprim sulfamethoxazole (in the absence of hydroxychloroquine). Subsequently he underwent aortic valve replacement. The particularities of this case consisted in atypical clinical manifestations, the absence of fever and epidemiological context suggestive for Q fever.


2017 ◽  
Vol 80 (4) ◽  
pp. 561-566 ◽  
Author(s):  
Mohammad M. Obaidat ◽  
Gilbert J. Kersh

ABSTRACT This large-scale cross-sectional study was conducted to determine the prevalence, geographical distribution, and risk factors for the presence of antibodies against Coxiella burnetii in bulk tank milk derived from dairy cattle, sheep, and goats in Jordan. Bulk milk samples were collected from 78 dairy cattle, 48 sheep, and 23 goat farms from various places in Jordan according to the density of these animal species in each region of the country. The samples were tested for C. burnetii antibodies using the CHEKIT Q-Fever Antibody ELISA kit. A standardized questionnaire was also used to collect data from each farm to identify and rank the risk factors for the presence of C. burnetii antibodies. The results revealed that 62.9% (95% confidence interval: 55.1 to 70.0%) of the tested ruminant farms were positive for C. burnetii antibodies. Positive results were obtained from 70.9% (60.6 to 79.5%) of dairy cattle farms, 52.1% (38.3 to 65.5%) of sheep farms, and 56.0% (37.1 to 73.3%) of goat farms. Six factors were associated with the presence of these antibodies on cattle farms, and five factors were associated with these antibodies on sheep and goat farms (chi-square test). The multivariate logistic regression model revealed that large dairy cattle farms, farms that add new animals to the herd, farms that infrequently clean the feeders, and farms in particular areas are 28.6, 19.9, 8.0, and 6.4 times more likely, respectively, to have animals with C. burnetii antibodies. Sheep and goat farms that mix their animals with those from other farms, graze more than 5 km, and infrequently sanitize the feeders were 8.0, 0.06, and 13.6 times more likely, respectively, to have animals with C. burnetii antibodies. These data reveal the widespread exposure of Jordanian ruminants to C. burnetii and suggest a high risk for public health.


2020 ◽  
Vol 25 (9) ◽  
pp. 1038-1046
Author(s):  
Eun Jeong Cho ◽  
Ashwini K. Devkota ◽  
Gabriel Stancu ◽  
Ramakrishna Edupunganti ◽  
Ginamarie Debevec ◽  
...  

Hypoxic solid tumors induce the stabilization of hypoxia-inducible factor 1 alpha (HIF1α), which stimulates the expression of many glycolytic enzymes and hypoxia-responsive genes. A high rate of glycolysis supports the energetic and material needs for tumors to grow. Fructose-1,6-bisphosphate aldolase A (ALDOA) is an enzyme in the glycolytic pathway that promotes the expression of HIF1α. Therefore, inhibition of ALDOA activity represents a potential therapeutic approach for a range of cancers by blocking two critical cancer survival mechanisms. Here, we present a luminescence-based strategy to determine ALDOA activity. The assay platform was developed by integrating a previously established ALDOA activity assay with a commercial NAD/NADH detection kit, resulting in a significant (>12-fold) improvement in signal/background (S/B) compared with previous assay platforms. A screening campaign using a mixture-based compound library exhibited excellent statistical parameters of Z′ (>0.8) and S/B (~20), confirming its robustness and readiness for high-throughput screening (HTS) application. This assay platform provides a cost-effective method for identifying ALDOA inhibitors using a large-scale HTS campaign.


2019 ◽  
Author(s):  
Rania Francis ◽  
Maxime Mioulane ◽  
Marion Le Bideau ◽  
Marie-Charlotte Mati ◽  
Pierre-Edouard Fournier ◽  
...  

AbstractQ fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although the current serological and molecular detection tools enable a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to evaluate their antibiotic susceptibility and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers as it requires biosafety-level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel high content screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically-designed algorithms targeting cytopathic effects. This method was more efficient than the shell-vial assay when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% vs 78% for shell-vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% vs 5% for shell-vial). In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell-vial culture and quantitative PCR. Overall, this high-throughput HCS system paves the way to the development of improved cell culture isolation of human viruses.


2012 ◽  
Vol 17 (6) ◽  
pp. 38-40
Author(s):  
A. A. Nafeev ◽  
V. V. Bezik

Coxiellosis (Q-fever) is a systemic natural focal zooanthroponosis disease, which has a very wide geographical area. A variety of clinical manifestations and the absence of pathognomonic signs of illness forces to use for making a definite diagnosis complex of techniques including epizootological, epidemiological data, clinical manifestations, and certainly the results of several serological tests designed to detect antibodies to Coxiella burnetii. There is presented a description of an acute case of Q - fever in a child in the subject, characterized as trouble-free of Coxiellosis from 70-ies of the last century.


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