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1.
Georgian Med News ; (311): 68-73, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33814394

RESUMEN

The aim - in the given research, the difficulties in interpreting the study results of oxidative homeostasis of oral fluid are analyzed. Changes in the total antioxidant activity of blood and saliva can be multidirectional - an increase or decrease in the oral fluid indicator and a reduction in the parameter of blood plasma can be recorded. To resolve the emerging difficulties, there was proposed a parallel assessment of the dynamics of changes in the total antioxidant activity of blood plasma and oral fluid in the patients of 4 groups with nosological forms of fundamentally different in the distribution and localization of the pathological process, which include: phlegmons of the maxillofacial region, partial absence of teeth, type 2 diabetes mellitus and the pelvic inflammatory diseases. As a result of the conducted studies, it was shown that a simultaneous decrease in the total antioxidant activity of blood plasma and oral fluid was attributable to the chronic long-term somatic diseases of a systemic character with a significant metabolic disorder, such as type 2 diabetes mellitus. A decrease in the total antioxidant activity of blood plasma and the unchanged oral fluid index was characteristic of somatic diseases of limited prevalence without affection of the maxillofacial region's tissues. In our case, such an example was a chronic inflammatory disease of the uterus with a combined course of bilateral salpingoophoritis. An increase in the oral fluid's total antioxidant activity on the background of a normal or even slightly reduced level of the antioxidant potential of blood plasma was characteristic of dental diseases. The latter situation was most likely for the dental profile diseases, in which damage to the oral tissues can provoke the leaching of cellular contents, including endogenous antioxidants or other components of regenerative activity in the oral fluid. Herein, changes in the antioxidant activity of blood plasma may reflect the prevalence of a pathological process at the systemic level or its limitation only to the dentoalveolar system's tissues and elements. As an example of such a situation, the patients with phlegmon of the maxillofacial region or patients with partial absence of teeth can be cited.


Asunto(s)
Antioxidantes , Diabetes Mellitus Tipo 2 , Femenino , Humanos , Oxidación-Reducción , Estrés Oxidativo , Saliva
2.
Biosens Bioelectron ; 180: 113111, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33743492

RESUMEN

Significant barriers to the diagnosis of latent and acute SARS-CoV-2 infection continue to hamper population-based screening efforts required to contain the COVID-19 pandemic in the absence of widely available antiviral therapeutics or vaccines. We report an aptamer-based SARS-CoV-2 salivary antigen assay employing only low-cost reagents ($3.20/test) and an off-the-shelf glucometer. The test was engineered around a glucometer as it is quantitative, easy to use, and the most prevalent piece of diagnostic equipment globally, making the test highly scalable with an infrastructure that is already in place. Furthermore, many glucometers connect to smartphones, providing an opportunity to integrate with contact tracing apps, medical providers, and electronic health records. In clinical testing, the developed assay detected SARS-CoV-2 infection in patient saliva across a range of viral loads - as benchmarked by RT-qPCR - within 1 h, with 100% sensitivity (positive percent agreement) and distinguished infected specimens from off-target antigens in uninfected controls with 100% specificity (negative percent agreement). We propose that this approach provides an inexpensive, rapid, and accurate diagnostic for distributed screening of SARS-CoV-2 infection at scale.


Asunto(s)
Antígenos Virales/análisis , Técnicas Biosensibles/métodos , /diagnóstico , Pruebas en el Punto de Atención , Saliva/virología , Adulto , Femenino , Humanos , Masculino , Fosfoproteínas/análisis , Técnica SELEX de Producción de Aptámeros , Sensibilidad y Especificidad , Glicoproteína de la Espiga del Coronavirus/análisis
3.
Emerg Infect Dis ; 27(4): 1146-1150, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33754989

RESUMEN

The expense of saliva collection devices designed to stabilize severe acute respiratory syndrome coronavirus 2 RNA is prohibitive to mass testing. However, virus RNA in nonsupplemented saliva is stable for extended periods and at elevated temperatures. Simple plastic tubes for saliva collection will make large-scale testing and continued surveillance easier.


Asunto(s)
/métodos , ARN Viral , Saliva/virología , /diagnóstico , /virología , Creación de Capacidad/métodos , Humanos , Estabilidad del ARN , ARN Viral/aislamiento & purificación , ARN Viral/fisiología , Reproducibilidad de los Resultados , Asignación de Recursos , /aislamiento & purificación , Manejo de Especímenes/economía , Manejo de Especímenes/instrumentación , Manejo de Especímenes/métodos
4.
Braz Oral Res ; 35: e032, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33729277

RESUMEN

This study tested the null hypothesis that antihistamine-containing syrup does not change salivary metabolites in vitro and in vivo. For the in vitro experiments, saliva from 10 volunteers was mixed with a syrup or pill suspension of loratadine (1 mg/ml Claritin®, Schering-Plough, Rio de Janeiro, Brazil). For the in vivo experiment, 10 volunteers performed a mouth rinse with 10 mL of antihistamine syrup (Claritin®; Schering-Plough, Rio de Janeiro, Brazil) for 20 seconds and then discarded the rinse water. After 20 seconds, 5 mL of unstimulated whole saliva was spit into a plastic tube kept on ice. The protein profile of in vitro and in vivo experiments was analyzed using 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The samples were also analyzed by nuclear magnetic resonance (NMR) spectroscopy, followed by Principal Component Analysis and Wilcoxon test (p < 0.05). There were differences in salivary metabolites after syrup interaction. The salivary concentrations of acetate, n-caproate, arginine, glutamate, and lysine among other metabolites were reduced with the syrup in both in vivo and in vitro experiments (p < 0.05), but no differences were observed when the pill suspension was used (p > 0.05). Similar changes in metabolite profiles were observed in both in vitro and in vivo experiments. Electrophoresis revealed no difference in the salivary protein pattern. The null hypothesis was rejected because the intake of syrup medicine changes the salivary composition and influences oral homeostasis and susceptibility to oral diseases.


Asunto(s)
Saliva , Proteínas y Péptidos Salivales , Brasil , Electroforesis en Gel de Poliacrilamida , Antagonistas de los Receptores Histamínicos , Humanos
5.
J Oral Facial Pain Headache ; 35(1): 54-61, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33730127

RESUMEN

AIMS: To clarify the mechanisms of hypersensitivity and spontaneous pain in intraoral structures in rats with diabetes mellitus (DM) accompanied by reduced saliva. METHODS: Adult male Sprague-Dawley rats received a single injection of streptozocin (50 mg/kg) to induce DM. Saliva volume, intraoral hypersensitivity to menthol and capsaicin solutions, and head-withdrawal thresholds (HWTs) to noxious heat and mechanical stimulation of the tongue and whisker pad were measured. RESULTS: On day 7 after streptozocin injection, rats with DM had a significantly reduced spontaneous saliva volume, polydipsia, capsaicin aversion of the intraoral mucosa, and a reduced HWT to noxious mechanical stimulation of the whisker pad skin. The HWT to noxious mechanical stimulation of the tongue reduced further on day 14 after streptozocin injection. These symptoms are similar to the orofacial and intraoral complaints of patients with DM. Meanwhile, reduction of HWT to noxious heat stimulation of the tongue and whisker pad were not observed. These results indicate that spontaneous intraoral mucosal pain and mechanical facial hypersensitivity are antecedent symptoms before mechanical hypersensitivity of the tongue. CONCLUSION: The mechanisms of saliva reduction, spontaneous intraoral mucosa pain, and mechanical hypersensitivity of intraoral and facial structures induced by DM involve both peripheral and autonomic neuropathies. Tongue hypersensitivity to noxious mechanical stimulation might be aggravated by xerostomia.


Asunto(s)
Diabetes Mellitus , Xerostomía , Animales , Humanos , Hiperalgesia , Masculino , Mucosa Bucal , Ratas , Ratas Sprague-Dawley , Saliva , Lengua , Xerostomía/inducido químicamente
6.
Front Public Health ; 9: 569315, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33718315

RESUMEN

Background: The COVID-19 pandemic has led to an urgent requirement for novel diagnostic tests that determine infection with SARS-CoV-2 and the development of an immune response against it. The perspective of end users on the characteristics and clinical use of these assays has not been previously considered. Methods: We surveyed 17,186 health care professions (HCPs) in 29 countries to gauge opinion on the design, use, diagnostic impact and diagnostic accuracy of COVID-19 tests. Results were correlated with national statistics on the burden of disease and testing in individual countries. Results: HCPs overwhelmingly recognized the importance of COVID-19 tests but 37.1% were unsure of the appropriate timing of investigations relative to disease symptoms. Confidence in the diagnostic accuracy of assays varied inversely with COVID-19-related mortality in individual countries but had no relationship with the total number of tests performed. There was global consensus that the most important impact of positive antigen and antibody testing was confidence in returning to work following recovery. Saliva was the preferred sampling fluid for COVID-19 diagnostic tests in all groups surveyed. Conclusions: HCP input can ensure novel assays are fit for purpose in varied global health care settings, but HCPs may require support to effectively use novel diagnostics thus minimizing waste when supplies are limited.


Asunto(s)
Salud Global , Personal de Salud/estadística & datos numéricos , Adulto , /mortalidad , Femenino , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Saliva
7.
Swiss Dent J ; 131(3): 253-255, 2021 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-33666388

RESUMEN

In addition to the tasks of pre-digestion, buffering of food acids, protection and remineralisation of tooth substance and moistening of the oral mucosa, saliva also plays an important role in wound healing, and a reduced flow rate represents a serious problem for patients. The saliva flow rate measurement (sialometry) on patients before radiation therapy should take place as a routine procedure, the costs are covered by the statutory health insurance. Due to the increased risk of caries in patients with reduced salivary flow, optimum oral hygiene and professional prophylactic care are crucial. In addition to nutritional guidance of the patient, saliva substitutes and medicinal salivary stimulants are available on the market. Saliva analysis has recently become more of public interest, especially in the field of personalized medicine as various drugs, viruses and DNA can be detected in saliva.


Asunto(s)
Caries Dental , Diente , Humanos , Mucosa Bucal , Higiene Bucal , Saliva
8.
Sensors (Basel) ; 21(5)2021 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-33652955

RESUMEN

Saliva can be used for health monitoring with non-invasive wearable systems. Such devices, including electrochemical sensors, may provide a safe, fast, and cost-efficient way of detecting target ions. Although salivary ions are known to reflect those in blood, no available clinical device can detect essential ions directly from saliva. Here, we introduce an all-solid-state, flexible film sensor that allows highly accurate detection of sodium levels in saliva, comparable to those in blood. The wireless film sensor system can successfully measure sodium ions from a small volume of infants' saliva (<400 µL), demonstrating its potential as a continuous health monitor. This study includes the structural characterization and error analysis of a carbon/elastomer-based ion-selective electrode and a reference electrode to confirm the signal reliability. The sensor, composed of a pair of the electrodes, shows good sensitivity (58.9 mV/decade) and selectivity (log K = -2.68 for potassium), along with a broad detection range of 5 × 10-5 ≈ 1 M with a low detection limit of 4.27 × 10-5 M. The simultaneous comparison between the film sensor and a commercial electrochemical sensor demonstrates the accuracy of the flexible sensor and a positive correlation in saliva-to-blood sodium levels. Collectively, the presented study shows the potential of the wireless ion-selective sensor system for a non-invasive, early disease diagnosis with saliva.


Asunto(s)
Técnicas Biosensibles , Sodio , Electrodos , Humanos , Lactante , Electrodos de Ion-Selectivo , Iones , Reproducibilidad de los Resultados , Saliva
9.
Sci Rep ; 11(1): 4943, 2021 03 02.
Artículo en Inglés | MEDLINE | ID: mdl-33654146

RESUMEN

The pandemic of COVID-19 is continuously spreading, becoming a worldwide emergency. Early and fast identification of subjects with a current or past infection must be achieved to slow down the epidemiological widening. Here we report a Raman-based approach for the analysis of saliva, able to significantly discriminate the signal of patients with a current infection by COVID-19 from healthy subjects and/or subjects with a past infection. Our results demonstrated the differences in saliva biochemical composition of the three experimental groups, with modifications grouped in specific attributable spectral regions. The Raman-based classification model was able to discriminate the signal collected from COVID-19 patients with accuracy, precision, sensitivity and specificity of more than 95%. In order to translate this discrimination from the signal-level to the patient-level, we developed a Deep Learning model obtaining accuracy in the range 89-92%. These findings have implications for the creation of a potential Raman-based diagnostic tool, using saliva as minimal invasive and highly informative biofluid, demonstrating the efficacy of the classification model.


Asunto(s)
/diagnóstico , Saliva/química , Espectrometría Raman/métodos , Anciano , Anciano de 80 o más Años , Anticuerpos Antivirales/análisis , Comorbilidad , Biología Computacional , Aprendizaje Profundo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Distribución Normal , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
11.
Methods Mol Biol ; 2265: 429-445, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33704732

RESUMEN

Canine oral melanoma (COM) is a common oral cancer with aggressiveness and high metastasis. A tumor in a dog's mouth makes it difficult to be observed at the early-clinical stage. Salivary biomarkers may be useful for early detection, prognosis, and monitoring of therapies. In addition, salivary collection is a simple and non-invasive technique. The present study describes how to identify salivary biomarkers in COM using matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) approaches. Western blot analysis has been used to confirm the protein expression. The sequence of expressed protein from western blot has been verified by LC-MS/MS.


Asunto(s)
Enfermedades de los Perros/metabolismo , Melanoma , Neoplasias de la Boca , Proteínas de Neoplasias/metabolismo , Saliva/metabolismo , Proteínas y Péptidos Salivales/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Animales , Perros , Melanoma/metabolismo , Melanoma/veterinaria , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/veterinaria
12.
Sci Rep ; 11(1): 6264, 2021 03 17.
Artículo en Inglés | MEDLINE | ID: mdl-33731722

RESUMEN

Many educational institutions have partially or fully closed all operations to cope with the challenges of the ongoing COVID-19 pandemic. In this paper, we explore strategies that such institutions can adopt to conduct safe reopening and resume operations during the pandemic. The research is motivated by the University of Illinois at Urbana-Champaign's (UIUC's) SHIELD program, which is a set of policies and strategies, including rapid saliva-based COVID-19 screening, for ensuring safety of students, faculty and staff to conduct in-person operations, at least partially. Specifically, we study how rapid bulk testing, contact tracing and preventative measures such as mask wearing, sanitization, and enforcement of social distancing can allow institutions to manage the epidemic spread. This work combines the power of analytical epidemic modeling, data analysis and agent-based simulations to derive policy insights. We develop an analytical model that takes into account the asymptomatic transmission of COVID-19, the effect of isolation via testing (both in bulk and through contact tracing) and the rate of contacts among people within and outside the institution. Next, we use data from the UIUC SHIELD program and 85 other universities to estimate parameters that describe the analytical model. Using the estimated parameters, we finally conduct agent-based simulations with various model parameters to evaluate testing and reopening strategies. The parameter estimates from UIUC and other universities show similar trends. For example, infection rates at various institutions grow rapidly in certain months and this growth correlates positively with infection rates in counties where the universities are located. Infection rates are also shown to be negatively correlated with testing rates at the institutions. Through agent-based simulations, we demonstrate that the key to designing an effective reopening strategy is a combination of rapid bulk testing and effective preventative measures such as mask wearing and social distancing. Multiple other factors help to reduce infection load, such as efficient contact tracing, reduced delay between testing and result revelation, tests with less false negatives and targeted testing of high-risk class among others. This paper contributes to the nascent literature on combating the COVID-19 pandemic and is especially relevant for educational institutions and similarly large organizations. We contribute by providing an analytical model that can be used to estimate key parameters from data, which in turn can be used to simulate the effect of different strategies for reopening. We quantify the relative effect of different strategies such as bulk testing, contact tracing, reduced infectivity and contact rates in the context of educational institutions. Specifically, we show that for the estimated average base infectivity of 0.025 ([Formula: see text]), a daily number of tests to population ratio T/N of 0.2, i.e., once a week testing for all individuals, is a good indicative threshold. However, this test to population ratio is sensitive to external infectivities, internal and external mobilities, delay in getting results after testing, and measures related to mask wearing and sanitization, which affect the base infection rate.


Asunto(s)
/prevención & control , Pandemias/prevención & control , Instituciones Académicas/normas , Universidades/normas , Enfermedades Asintomáticas , Simulación por Computador , Trazado de Contacto/métodos , Humanos , Saliva/virología
13.
Medicine (Baltimore) ; 100(10): e24742, 2021 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-33725829

RESUMEN

ABSTRACT: Oral microbiota has been implicated in pathogenesis of recurrent aphthous stomatitis (RAS), which is a common mucosal disorder with unclear etiology. This study has explored the association between oral microbiota disorder and RAS in high-risk young female population.Forty-five young females were enrolled, including 24 RAS patients and 21 healthy individuals. Oral microbiome was analyzed by Illumina Miseq sequencing.Oral microbiota associated with RAS was characterized by the lower alpha-diversity indices (Chao1 and ACE). Several infectious pathogens increased in RAS, such as genera Actinobacillus, Haemophilus, Prevotella and Vibrio. The PICRUSt analysis indicated that the oral microbiota might be related with the up-regulation of genes involving infectious and neurodegenerative diseases, environmental adaptation, the down-regulation of genes involving basal metabolism, such as carbohydrate, energy, and amino acid metabolism.This study indicated that oral microbiota may play a significant role in RAS development.


Asunto(s)
Microbiota , Mucosa Bucal/microbiología , Estomatitis Aftosa/microbiología , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Recurrencia , Saliva/microbiología , Adulto Joven
14.
Medicine (Baltimore) ; 100(10): e24924, 2021 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-33725852

RESUMEN

ABSTRACT: Orthodontic treatment can lead to microbial-induced gingival inflammation and aseptic periodontal inflammations. The aim of this study was to investigate the relationship between salivary pro-inflammatory cytokines levels with gingival health status and oral microbe loads among patients undergoing orthodontic treatment.The present investigation was a cross-sectional study among a sample of 111 consecutive orthodontic patients (mean age 18.4 ±â€Š4.4 years). Clinical examinations were conducted to assess the gingival health status employing the Modified Gingival Index, Gingival Bleeding Index, and Plaque Index. Salivary microbiological assessments of total aerobic and anaerobic bacteria count, streptococci count, and lactobacilli count were undertaken. Saliva immunological assessments included Interleukin-1Beta (IL-1ß) and macrophage migration inhibitory factor (MIF) ELISA assays.The mean ±â€Šstandard deviation of salivary IL-1ß was 83.52 ±â€Š85.62 pg/ml and MIF was 4.12 ±â€Š0.96 ng/ml. Moderate positive correlations were found between salivary IL-1ß levels and total aerobic and anaerobic bacteria count, streptococci count, and lactobacilli count (r = 0.380-0.446, P < .001), and weak positive correlations between salivary MIF levels and total salivary aerobic and anaerobic bacteria counts (r = 0.249-0.306, P < .01) were observed. A positive correlation was found between salivary IL-1ß levels and Bleeding Index (r = 0.216, P < .05).The level of salivary IL-1ß positively correlates with oral bacterial load among orthodontic patients; the relationship between inflammatory cytokines and oral microflora deserved further study.


Asunto(s)
Gingivitis/diagnóstico , Interleucina-1beta/análisis , Aparatos Ortodóncicos/efectos adversos , Saliva/química , Adolescente , Carga Bacteriana , Estudios Transversales , Femenino , Encía/inmunología , Encía/microbiología , Gingivitis/inmunología , Gingivitis/microbiología , Gingivitis/prevención & control , Humanos , Interleucina-1beta/inmunología , Oxidorreductasas Intramoleculares/análisis , Oxidorreductasas Intramoleculares/inmunología , Factores Inhibidores de la Migración de Macrófagos/análisis , Factores Inhibidores de la Migración de Macrófagos/inmunología , Masculino , Microbiota/inmunología , Antisépticos Bucales/administración & dosificación , Adulto Joven
15.
Sci Rep ; 11(1): 5448, 2021 03 09.
Artículo en Inglés | MEDLINE | ID: mdl-33750853

RESUMEN

To safely re-open economies and prevent future outbreaks, rapid, frequent, point-of-need, SARS-CoV-2 diagnostic testing is necessary. However, existing field-deployable COVID-19 testing methods require the use of uncomfortable swabs and trained providers in PPE, while saliva-based methods must be transported to high complexity laboratories for testing. Here, we report the development and clinical validation of High-Performance Loop-mediated isothermal Amplification (HP-LAMP), a rapid, saliva-based, SARS-CoV-2 test with a limit of detection of 1.4 copies of virus per µl of saliva and a sensitivity and specificity with clinical samples of > 96%, on par with traditional RT-PCR based methods using swabs, but can deliver results using only a single fluid transfer step and simple heat block. Testing of 120 patient samples in 40 pools comprised of 5 patient samples each with either all negative or a single positive patient sample was 100% accurate. Thus, HP-LAMP may enable rapid and accurate results in the field using saliva, without need of a high-complexity laboratory.


Asunto(s)
/diagnóstico , Saliva/virología , /virología , Humanos , Límite de Detección , Técnicas de Diagnóstico Molecular , Nasofaringe/virología , Técnicas de Amplificación de Ácido Nucleico , ARN Viral/metabolismo , Sensibilidad y Especificidad , Temperatura
16.
Emerg Infect Dis ; 27(4)2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33755009

RESUMEN

We analyzed feasibility of pooling saliva samples for severe acute respiratory syndrome coronavirus 2 testing and found that sensitivity decreased according to pool size: 5 samples/pool, 7.4% reduction; 10 samples/pool, 11.1%; and 20 samples/pool, 14.8%. When virus prevalence is >2.6%, pools of 5 require fewer tests; when <0.6%, pools of 20 support screening strategies.


Asunto(s)
/métodos , Saliva/virología , Manejo de Especímenes/métodos , /diagnóstico , Creación de Capacidad/métodos , Asignación de Recursos para la Atención de Salud , Humanos , Límite de Detección , Asignación de Recursos/métodos , Sensibilidad y Especificidad , Estados Unidos
17.
BMJ Open Respir Res ; 8(1)2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33627333

RESUMEN

BACKGROUND: An outbreak of novel coronavirus (SARS-CoV-2)-associated respiratory infectious diseases (COVID-19) emerged in 2019 and has spread rapidly in humans around the world. The demonstration of in vitro infectiousness of respiratory specimens is an informative surrogate for SARS-CoV-2 transmission from patients with COVID-19; accordingly, viral isolation assays in cell culture are an important aspect of laboratory diagnostics for COVID-19. METHODS: We developed a simple and rapid protocol for isolating SARS-CoV-2 from respiratory specimens using VeroE6/TMPRSS2 cells, a cell line that is highly susceptible to the virus. We also investigated a correlation between isolation of SARS-CoV-2 and viral load detected by real-time RT-PCR (rRT-PCR) using N2 primer/probe set that has been developed for testing of COVID-19 in Japan. RESULTS: The SARS-CoV-2 isolation protocol did not require blind passage of inoculated cells and yielded the results of viral isolation within 7 days after inoculation. Specimens with cycle threshold (Ct) values of <20.2, determined by rRT-PCR, were predicted to be isolation-positive. On the other hand, 6.9% of specimens with Ct values >35 were virus isolation-positive, indicating that low viral loads (high Ct values) in upper respiratory specimens do not always indicate no risk of containing transmissible virus. CONCLUSION: In combination with rRT-PCR, the SARS-CoV-2 isolation protocol provides a means for assessing the potential risk of transmissible virus in upper respiratory specimens.


Asunto(s)
/transmisión , /patogenicidad , Animales , Línea Celular , Chlorocebus aethiops , Efecto Citopatogénico Viral , Humanos , Cavidad Nasal/virología , Nasofaringe/virología , Saliva/virología , Serina Endopeptidasas/genética , Manejo de Especímenes , Células Vero
18.
Anal Methods ; 13(7): 874-883, 2021 02 25.
Artículo en Inglés | MEDLINE | ID: mdl-33576354

RESUMEN

Using the children's toy, Shrinky-Dink©, we present an aptamer-based electrochemical (E-AB) assay that recognizes the spike protein of SARS-CoV-2 in saliva for viral infection detection. The low-cost electrodes are implementable at population scale and demonstrate detection down to 1 ag mL-1 of the S1 subunit of the spike protein.


Asunto(s)
Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Poliestirenos/química , Saliva/química , Glicoproteína de la Espiga del Coronavirus/análisis , Aptámeros de Nucleótidos/química , Técnicas Electroquímicas/instrumentación , Electrodos , Oro/química , Humanos , Límite de Detección , Dominios Proteicos , Glicoproteína de la Espiga del Coronavirus/química
19.
Viruses ; 13(2)2021 02 06.
Artículo en Inglés | MEDLINE | ID: mdl-33562073

RESUMEN

The contemporary surge in metagenomic sequencing has transformed knowledge of viral diversity in wildlife. However, evaluating which newly discovered viruses pose sufficient risk of infecting humans to merit detailed laboratory characterization and surveillance remains largely speculative. Machine learning algorithms have been developed to address this imbalance by ranking the relative likelihood of human infection based on viral genome sequences, but are not yet routinely applied to viruses at the time of their discovery. Here, we characterized viral genomes detected through metagenomic sequencing of feces and saliva from common vampire bats (Desmodus rotundus) and used these data as a case study in evaluating zoonotic potential using molecular sequencing data. Of 58 detected viral families, including 17 which infect mammals, the only known zoonosis detected was rabies virus; however, additional genomes were detected from the families Hepeviridae, Coronaviridae, Reoviridae, Astroviridae and Picornaviridae, all of which contain human-infecting species. In phylogenetic analyses, novel vampire bat viruses most frequently grouped with other bat viruses that are not currently known to infect humans. In agreement, machine learning models built from only phylogenetic information ranked all novel viruses similarly, yielding little insight into zoonotic potential. In contrast, genome composition-based machine learning models estimated different levels of zoonotic potential, even for closely related viruses, categorizing one out of four detected hepeviruses and two out of three picornaviruses as having high priority for further research. We highlight the value of evaluating zoonotic potential beyond ad hoc consideration of phylogeny and provide surveillance recommendations for novel viruses in a wildlife host which has frequent contact with humans and domestic animals.


Asunto(s)
Quirópteros/virología , Virus/aislamiento & purificación , Zoonosis/virología , Animales , Reservorios de Enfermedades/veterinaria , Reservorios de Enfermedades/virología , Heces/virología , Genoma Viral/genética , Humanos , Aprendizaje Automático , Metagenómica , Filogenia , Virus de la Rabia/clasificación , Virus de la Rabia/genética , Virus de la Rabia/aislamiento & purificación , Saliva/virología , Virus/clasificación , Virus/genética
20.
Animal ; 15(2): 100095, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33573980

RESUMEN

Optimal management of gilt reproduction requires oestrus synchronization. Hormonal treatments are used for this purpose, but there is a growing demand for non-hormonal alternatives, especially in organic farms. The boar effect is an important alternative opportunity to induce and synchronize oestrus without hormones. Before puberty, gilts exhibit a 'waiting period' during which boar exposure could induce and synchronize the first ovulation. We searched for salivary biomarkers of this period of boar effect receptivity to improve detection of the gilts to stimulate with the perspective of enhancing the efficacy of the boar effect. Saliva samples were collected from 30 Large-White×Landrace crossbred gilts between 140 and 175 days of age. Gilts were exposed twice a day to a boar and subjected to oestrus detection from 150 to 175 days of age. Among the 30 gilts, 10 were detected in oestrus 4 to 7 days after the first introduction of the boar and were considered receptive to the boar effect, 14 were detected in oestrus more than 8 days after first boar contact, and six did not show oestrus and were considered non-receptive. Saliva samples from six receptive and six non-receptive gilts were analyzed for steroidome and for metabolome using gas chromatography coupled to tandem mass spectrometry and 1H nuclear magnetic resonance spectroscopy, respectively. Four saliva samples per gilt were analyzed: 25 days and 11 days before boar introduction, the day of boar introduction, 3 days later for receptive gilts or 7 days later for non-receptive gilts. Twenty-nine steroids and 31 metabolites were detected in gilt saliva. Salivary concentrations of six steroids and three metabolites were significantly different between receptive and non-receptive gilts: progesterone and glycolate 25 days before boar introduction, 3α5ß20α- and 3ß5α20ß-hexahydroprogesterone, dehydroepiandrosterone, androstenediol, succinate, and butyrate 11 days before boar introduction, and 3ß5α-tetrahydroprogesterone on the day of boar introduction. Thus, nine potential salivary biomarkers of boar effect receptivity were identified in our experimental conditions. Further studies with higher numbers of gilts and salivary sampling points are necessary to ascertain their reliability.


Asunto(s)
Saliva , Maduración Sexual , Animales , Biomarcadores , Femenino , Cromatografía de Gases y Espectrometría de Masas/veterinaria , Masculino , Metaboloma , Reproducibilidad de los Resultados , Porcinos
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