143 Non-invasive skin sample collection: Comparing D-squame and the Smart Sticker

Background:Pyogenic spondylodiscitis (SPD) is a serious infection of an intervertebral disc and/or adjacent vertebrae, that remains a topical problem in rheumatological practice. Early diagnosis and treatment are the only guarantees of a favorable outcome. Clinicians must strive to isolate the responsible bacteria in order to adapt the treatment, and thus reduce the risk of resistance and complications due to SPD itself, but also to the multiplication of probabilistic treatments.Objectives:Our aim was to study the contribution of the different microbiological and anatomopathological examinations in the diagnosis of pyogenic SPD.Methods:It was a descriptive study in a single rheumatology department. Data were collected retrospectively from observations of patients hospitalized in the past 20 years who have been diagnosed with pyogenic SPD. We excluded cases of tuberculous and brucellar SPD from our study because of their completely different histological and microbiological profiles.Results:Twenty-two cases of pyogenic SPD were collected (14M/ 8F). The mean age of the population was 55.9 years [29,80]. A bacteriological survey including at least one cytobacteriological examination of the urine (CBEU), chest X-rays and blood cultures allowed the identification of the bacteria in 16 cases (73%). The most common site were bacteria was identified was blood culture in 7 cases, skin sample and urine collection in 2 cases each. Disco-vertebral puncture and biopsy (DVPB) was performed in 19 patients when there was no bacteria identification and/or when diagnosis of infectious SPD persisted doubtful. On histopathological examination, were described: an infiltrate and/or inflammatory changes without specificity signs in 7 patients and an appearance of chronic pyogenic SPD very likely in 12 patients. Bacteriological study of DVPB fluid or paravertebral abscesses sample helped to isolate bacteria in 4 patients. DVPB or abscesses puncture were contributing by histological and/or bacteriological examination in 12 patients (63%).Infecting bacteria was identified in 14 patients (64%). Gram-negative bacilli (GNB) and staphylococcus aureus were the most frequent germs (7 cases each) including 2 cases of co-infection. GNBs were represented by: Escherichia Coli and Enterobacter Cloacae in 2 cases each, Proteus Mirabilis, Serratia Marcescens and Klebsiella oxytoca in 1 case each. Clostridium clostridioforme and Lactococcus cremoris were isolated in 1 case each. For patients whose etiological investigation remained negative, SPD diagnosis was retained based on imaging (MRI) guided by anamnestic, clinico-biological and histopathological arguments.Conclusion:SPD is a rare condition that needs to be treated rapidly. Once the diagnosis is suspected, bacteria must be isolated before starting any antibiotic therapy. Simple and non-invasive exams as blood cultures, CBUE and chest rays, should be undertaken first. In fact, these simple exams allowed a germ identification in 73% cases in our study. If doubt persist, DVPB could be contributive to the diagnosis.References:NoneDisclosure of Interests:None declared

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13-gene DNA Methylation Analysis from Oral Brushing: A Promising Non Invasive Tool in the Follow-up of Oral Cancer Patients

Journal of Clinical Medicine ◽

10.3390/jcm8122107 ◽

2019 ◽

Vol 8 (12) ◽

pp. 2107 ◽

Cited By ~ 4

Author(s):

Davide B. Gissi ◽

Achille Tarsitano ◽

Andrea Gabusi ◽

Roberto Rossi ◽

Giuseppe Attardo ◽

...

Keyword(s):

Dna Methylation ◽

Oral Cancer ◽

Cancer Treatment ◽

Sample Collection ◽

Methylation Analysis ◽

Non Invasive ◽

Locoregional Relapse ◽

Positive Score ◽

Dna Methylation Analysis

Background: This study aimed to evaluate the prognostic value of a non-invasive sampling procedure based on 13-gene DNA methylation analysis in the follow-up of patients previously treated for oral squamous cell carcinoma (OSCC). Methods: The study population included 49 consecutive patients treated for OSCC. Oral brushing sample collection was performed at two different times: before any cancer treatment in the tumor mass and during patient follow-up almost 6 months after OSCC treatment, within the regenerative area after OSCC resection. Each sample was considered positive or negative in relation to a predefined cut-off value. Results: Before any cancer treatment, 47/49 specimens exceeded the score and were considered as positive. Six months after OSCC resection, 16/49 specimens also had positive scores in the samples collected from the regenerative area. During the follow-up period, 7/49 patients developed locoregional relapse: 6/7 patients had a positive score in the regenerative area after OSCC resection. The presence of a positive score after oral cancer treatment was the most powerful variable related to the appearance of locoregional relapse. Conclusion: 13-gene DNA methylation analysis by oral brushing may have a clinical application as a prognostic non-invasive tool in the follow-up of patients surgically treated for OSCC.

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Factors influencing genotyping success and genotyping error rate of Eurasian otter (Lutra lutra) faeces collected in temperate Central Europe

European Journal of Wildlife Research ◽

10.1007/s10344-020-01444-4 ◽

2020 ◽

Vol 67 (1) ◽

Author(s):

Marcia Sittenthaler ◽

Eva Maria Schöll ◽

Christoph Leeb ◽

Elisabeth Haring ◽

Rosemarie Parz-Gollner ◽

...

Keyword(s):

Influential Factors ◽

Lutra Lutra ◽

Sample Treatment ◽

Success Rates ◽

Sample Collection ◽

Allelic Dropout ◽

Non Invasive ◽

Eurasian Otter ◽

Faecal Samples ◽

Elusive Species

AbstractThe use of non-invasively collected DNA source material for genetic and genomic applications is usually characterized by low target DNA concentration and quality, genotyping errors and cost-intensive lab procedures. However, for otters (Lutrinae) as elusive species of conservation concern, genetic non-invasive sampling has become an important tool to study their ecology and demography. To increase cost-efficiency of monitoring programmes and to promote the expansion of genomic approaches to non-invasive samples, we aimed to refine sample collection and preparation. Therefore, we examined the effects of intrinsic sample characteristics (including diet), environmental conditions in the field and sample treatment in the molecular laboratory on the success of genotyping and allelic dropout (ADO) rates using microsatellite markers in 1970 fresh Eurasian otter (Lutra lutra) scats. Using fresh samples only, we probably eliminated one of the most important impediments of genotyping DNA from otter faecal samples beforehand. But, we observed higher genotyping success and lower ADO rates for anal glad secretions and faecal samples containing high proportions of mucus. Moist conditions during sample collection may promote DNA degradation and PCR inhibition, leading to decreased genotyping success rates. ADO was further affected by the type of extraction kit. However, a high proportion of variance remaining unexplained by our models implied that additional parameters were acting (amount of PCR inhibitors, non-uniform distribution of intestinal cells, efficiency of PCRs, specific microclimate at marking sites). We summarized influential factors maximizing genotyping quality of otter scats and give recommendations for sample collection, storage and DNA extraction based on our results and current literature.

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Saliva Samples as A Tool to Study the Effect of Meal Timing on Metabolic And Inflammatory Biomarkers

Nutrients ◽

10.3390/nu12020340 ◽

2020 ◽

Vol 12 (2) ◽

pp. 340 ◽

Cited By ~ 2

Author(s):

Katharina Kessler ◽

Silke Hornemann ◽

Natalia Rudovich ◽

Daniela Weber ◽

Tilman Grune ◽

...

Keyword(s):

Metabolic Regulation ◽

Diurnal Variations ◽

Inflammatory Biomarkers ◽

Blood Levels ◽

Fat Intake ◽

Sample Collection ◽

High Fat ◽

Non Invasive ◽

Promising Tool ◽

Meal Timing

Meal timing affects metabolic regulation in humans. Most studies use blood samples for their investigations. Saliva, although easily available and non-invasive, seems to be rarely used for chrononutritional studies. In this pilot study, we tested if saliva samples could be used to study the effect of timing of carbohydrate and fat intake on metabolic rhythms. In this cross-over trial, 29 nonobese men were randomized to two isocaloric 4-week diets: (1) carbohydrate-rich meals until 13:30 and high-fat meals between 16:30 and 22:00 or (2) the inverse order of meals. Stimulated saliva samples were collected every 4 h for 24 h at the end of each intervention, and levels of hormones and inflammatory biomarkers were assessed in saliva and blood. Cortisol, melatonin, resistin, adiponectin, interleukin-6 and MCP-1 demonstrated distinct diurnal variations, mirroring daytime reports in blood and showing significant correlations with blood levels. The rhythm patterns were similar for both diets, indicating that timing of carbohydrate and fat intake has a minimal effect on metabolic and inflammatory biomarkers in saliva. Our study revealed that saliva is a promising tool for the non-invasive assessment of metabolic rhythms in chrononutritional studies, but standardisation of sample collection is needed in out-of-lab studies.

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Non-invasive sample collection for respiratory virus testing by multiplex PCR

Journal of Clinical Virology ◽

10.1016/j.jcv.2011.07.015 ◽

2011 ◽

Vol 52 (3) ◽

pp. 210-214 ◽

Cited By ~ 30

Author(s):

Anne J. Blaschke ◽

Mandy A. Allison ◽

Lindsay Meyers ◽

Margarita Rogatcheva ◽

Caroline Heyrend ◽

...

Keyword(s):

Multiplex Pcr ◽

Respiratory Virus ◽

Sample Collection ◽

Non Invasive ◽

Virus Testing

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COVID-19 salivary signature: diagnostic and research opportunities

Journal of Clinical Pathology ◽

10.1136/jclinpath-2020-206834 ◽

2020 ◽

pp. jclinpath-2020-206834 ◽

Cited By ~ 10

Author(s):

Dipak Sapkota ◽

Tine Merete Søland ◽

Hilde Kanli Galtung ◽

Lars Peter Sand ◽

Simone Giannecchini ◽

...

Keyword(s):

Disease Transmission ◽

Scientific Basis ◽

Healthcare Personnel ◽

Sample Collection ◽

Promising Alternative ◽

Invasive Approach ◽

Non Invasive ◽

Saliva Collection ◽

Personnel Safety ◽

Degrees Of Severity

The COVID-19 (caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)) epidemic started in Wuhan (Hubei Province, China) in mid-December 2019 and quickly spread across the world as a pandemic. As a key to tracing the disease and to implement strategies aimed at breaking the chain of disease transmission, extensive testing for SARS-CoV-2 was suggested. Although nasopharyngeal/oropharyngeal swabs are the most commonly used biological samples for SARS-CoV-2 diagnosis, they have a number of limitations related to sample collection and healthcare personnel safety. In this context, saliva is emerging as a promising alternative to nasopharyngeal/oropharyngeal swabs for COVID-19 diagnosis and monitoring. Saliva collection, being a non-invasive approach with possibility for self-collection, circumvents to a great extent the limitations associated with the use of nasopharyngeal/oropharyngeal swabs. In addition, various salivary biomarkers including the salivary metabolomics offer a high promise to be useful for better understanding of COVID-19 and possibly in the identification of patients with various degrees of severity, including asymptomatic carriers. This review summarises the clinical and scientific basis for the potential use of saliva for COVID-19 diagnosis and disease monitoring. Additionally, we discuss saliva-based biomarkers and their potential clinical and research applications related to COVID-19.

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Opportunities and challenges in metabarcoding approaches for helminth community identification in wild mammals

Parasitology ◽

10.1017/s0031182017000610 ◽

2017 ◽

Vol 145 (5) ◽

pp. 608-621 ◽

Cited By ~ 6

Author(s):

TUOMAS AIVELO ◽

ALAN MEDLAR

Keyword(s):

Statistical Analysis ◽

Bacterial Community ◽

Bacterial Community Composition ◽

Accurate Assessment ◽

Helminth Community ◽

Sample Collection ◽

Wild Mammals ◽

Non Invasive ◽

Community Identification ◽

Parasitological Study

SUMMARYDespite metabarcoding being widely used to analyse bacterial community composition, its application in parasitological research remains limited. What interest there has been has focused on previously intractable research settings where traditional methods are inappropriate, for example, in longitudinal studies and studies involving endangered species. In settings such as these, non-invasive sampling combined with metabarcoding can provide a fast and accurate assessment of component communities. In this paper we review the use of metabarcoding in the study of helminth communities in wild mammals, outlining the necessary procedures from sample collection to statistical analysis. We highlight the limitations of the metabarcoding approach and speculate on what type of parasitological study would benefit from such methods in the future.

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A robotic platform for fluidically-linked human body-on-chips experimentation

10.1101/569541 ◽

2019 ◽

Cited By ~ 1

Author(s):

Richard Novak ◽

Miles Ingram ◽

Susan Clauson ◽

Debarun Das ◽

Aaron Delahanty ◽

...

Keyword(s):

Human Body ◽

Experimental System ◽

Blood Substitute ◽

Sample Collection ◽

Human Organ ◽

Non Invasive ◽

Organ Specific ◽

Repeated Sampling ◽

Substitute Medium

Here we describe of an ‘Interrogator’ instrument that uses liquid-handling robotics, a custom software package, and an integrated mobile microscope to enable automated culture, perfusion, medium addition, fluidic linking, sample collection, andin situmicroscopic imaging of up to 10 Organ Chips inside a standard tissue culture incubator. The automated Interrogator platform maintained the viability and organ-specific functions of 8 different vascularized, 2-channel, Organ Chips (intestine, liver, kidney, heart, lung, skin, blood-brain barrier (BBB), and brain) for 3 weeks in culture when fluidically coupled through their endothelium-lined vascular channels using a common blood substitute medium. When an inulin tracer was perfused through the multi-organ Human Body-on-Chips (HuBoC) fluidic network, quantitative distributions of this tracer could be accurately predicted using a physiologically-based multi-compartmental reduced order (MCRO)in silicomodel of the experimental system derived from first principles. This automated culture platform enables non-invasive imaging of cells within human Organ Chips and repeated sampling of both the vascular and interstitial compartments without compromising fluidic coupling, which should facilitate future HuBoc studies and pharmacokinetics (PK) analysisin vitro.

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Sample Adequacy Controls for Infectious Disease Diagnosis by Oral Swabbing

10.1101/2020.07.17.20156489 ◽

2020 ◽

Author(s):

Meagan Deviaene ◽

Kris M. Weigel ◽

Rachel C Wood ◽

Angelique K Luabeya ◽

Lisa Jones-Engel ◽

...

Keyword(s):

Oral Cavity ◽

Cell Types ◽

Disease Diagnosis ◽

Oral Microbiota ◽

Sample Type ◽

Sample Collection ◽

Total N ◽

Oral Swab ◽

Non Invasive ◽

Negative Controls

Oral swabs are emerging as a non-invasive sample type for diagnosing infectious diseases including Ebola, tuberculosis (TB), and COVID-19. To assure proper sample collection, sample adequacy controls (SACs) are needed that detect substances indicative of samples collected within the oral cavity. This study evaluated two candidate SACs for this purpose. One detected representative oral microbiota (Streptococcus species DNA) and the other, human cells (human mitochondrial DNA, mtDNA). Quantitative PCR (qPCR) assays for the two target cell types were applied to buccal swabs (representing samples collected within the oral cavity) and hand swabs (representing improperly collected samples) obtained from 51 healthy U.S. volunteers. Quantification cycle (Cq) cutoffs that maximized Youdens index were established for each assay. The streptococcal target at a Cq cutoff of 34.9 had 99.0% sensitivity and specificity for oral swab samples, whereas human mtDNA perfectly distinguished between hand and mouth swabs with a Cq cutoff of 31.3. The human mtDNA test was then applied to buccal, tongue, and gum swabs that had previously been collected from TB patients and controls in South Africa, along with air swabs collected as negative controls (total N = 292 swabs from 71 subjects). Of these swabs, 287/292 (98%) exhibited the expected Cq values. In a paired analysis the three oral sites yielded indistinguishable amounts of human mtDNA, however PurFlockTM swabs collected slightly more human mtDNA than did OmniSwabsTM (p = 0.012). The results indicate that quantification of human mtDNA cannot distinguish swabs collected from different sites within the mouth. However, it can reliably distinguish oral swabs from swabs that were not used orally., which makes it a useful SAC for oral swab-based diagnosis.

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