scholarly journals Self-collected Saline Gargle Samples as an Alternative to Healthcare Worker Collected Nasopharyngeal Swabs for COVID-19 Diagnosis in Outpatients

2020 ◽  
Author(s):  
David M Goldfarb ◽  
Peter Tilley ◽  
Ghada N. Al-Rawahi ◽  
Jocelyn Srigley ◽  
Geoffrey Ford ◽  
...  
Keyword(s):  
Room Temperature ◽  
Positive Sample ◽  
Nasopharyngeal Swab ◽  
Molecular Testing ◽  
Sample Type ◽  
Mouth Rinse ◽  
Rna Detection ◽  
School Aged Children ◽  
Primary Sample ◽  
User Acceptability

Background: We assessed the performance, stability, and user acceptability of swab-independent self-collected saliva and saline mouth rinse/gargle sample types for the molecular detection of SARS-CoV-2 in adults and school-aged children. Methods: Outpatients who had recently been diagnosed with COVID-19 or were presenting with suspected COVID-19 were asked to have a nasopharyngeal swab collected and provide at least one self-collected sample type. A portion of participants were also asked about sample acceptability. Samples underwent molecular testing using multiple assays. Saline mouth rinse/gargle and saliva samples were tested daily at time zero, day one, and day 2 to assess nucleic acid stability at room temperature. Results: 50 participants (aged 4 to 71 years) were included; of these, 40 had at least one positive sample and were included in the primary sample yield analysis. Saline mouth rinse/gargle samples had a sensitivity of 98% (39/40) while saliva samples had a sensitivity of 79% (26/33). Both saline mouth rinse/gargle and saliva samples showed stable viral RNA detection after 2 days of room temperature storage. Mouth rinse/gargle samples had the highest (mean 4.9) and HCW-collected NP swabs had the lowest acceptability scores (mean 3.1). Conclusion: Saline mouth rinse/gargle samples demonstrated the highest combined user acceptability ratings and analytical performance when compared with saliva and HCW collected NP swabs. This sample type is a promising swab-independent option, particularly for outpatient self-collection in adults and school aged children.

scholarly journals Self-collected Saline Gargle Samples as an Alternative to Healthcare Worker Collected Nasopharyngeal Swabs for COVID-19 Diagnosis in Outpatients

2021 ◽  
Author(s):  
David M. Goldfarb ◽  
Peter Tilley ◽  
Ghada N. Al-Rawahi ◽  
Jocelyn A. Srigley ◽  
Geoffrey Ford ◽  
...  
Keyword(s):  
Room Temperature ◽  
Positive Sample ◽  
Nasopharyngeal Swab ◽  
Sample Type ◽  
Mouth Rinse ◽  
Rna Detection ◽  
School Aged Children ◽  
Primary Sample ◽  
User Acceptability ◽  
Temperature Storage

Background: We assessed the performance, stability, and user acceptability of swab-independent self-collected saliva and saline mouth rinse/gargle sample types for the molecular detection of SARS-CoV-2 in adults and school-aged children. Methods: Outpatients who had recently been diagnosed with COVID-19 or were presenting with suspected COVID-19 were asked to have a nasopharyngeal swab collected and provide at least one self-collected sample type. Participants were also asked about sample acceptability using a five point Likert scale. For those previously diagnosed with COVID-19, all samples underwent real-time PCR testing using a lab-developed assay, and the majority were also tested using an FDA-authorized assay. For those presenting with suspect COVID-19, only those with a positive nasopharyngeal swab sample went on to have other samples tested. Saline mouth rinse/gargle and saliva samples were tested daily at time zero, day one, and day 2 to assess nucleic acid stability at room temperature. Results: 50 participants (aged 4 to 71 years) were included; of these, 40 had at least one positive sample and were included in the primary sample yield analysis. Saline mouth rinse/gargle samples had a sensitivity of 98% (39/40) while saliva samples had a sensitivity of 79% (26/33). Both saline mouth rinse/gargle and saliva samples showed stable viral RNA detection after 2 days of room temperature storage. Mouth rinse/gargle samples had the highest (mean 4.9) and HCW-collected NP swabs had the lowest acceptability scores (mean 3.1). Conclusion: Saline mouth rinse/gargle samples demonstrated the highest combined user acceptability ratings and analytical performance when compared with saliva and HCW collected NP swabs. This sample type is a promising swab-independent option, particularly for outpatient self-collection in adults and school aged children.

An ultrasensitive amplicon-based sequencing panel for noninvasive molecular testing of hematological malignancies using blood.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e19511-e19511
Author(s):  
Chae Yin Cher ◽  
Zi Yi Lim ◽  
Daryl Tan ◽  
Hae Tha Mya ◽  
Wichean Mongkonsritragoon ◽  
...  
Keyword(s):  
Hematological Malignancies ◽  
Clinical Decision Making ◽  
Myeloproliferative Neoplasms ◽  
Noise Removal ◽  
Molecular Testing ◽  
Sample Type ◽  
Monitoring Methods ◽  
Genomic Alterations ◽  
Alternative Source ◽  
Primary Sample

e19511 Background: Hematological malignancies, especially acute myeloid leukemia, myeloproliferative neoplasms, and myelodysplastic syndromes (MDS), present with distinct genomic alterations relevant to prognosis. Bone marrow (BM) is the primary sample source for genetic testing, but is obtained by an invasive biopsy procedure which carries risk of infection. An alternative non-invasive sample type to profile disease-relevant genomic alterations at diagnosis and subsequently, is desired for clinical decision making. An ultrasensitive sequencing panel was applied to peripheral blood (PB) as an alternative to BM, to demonstrate its utility for disease profiling and monitoring. Methods: A total of 27 pairs of matched PB and BM samples were obtained from patients with hematological malignancies including leukemia (n = 21), lymphoma (n = 4), multiple myeloma (n = 1) and MDS (n = 1). The sequencing panel covers 45 genes commonly mutated in myeloid and lymphoid malignancies. The panel is based on amplicon-sequencing and includes error correction for variant detection with allele frequency (VAF) as low as 0.1%. Optimized bioinformatics pipeline and in-house sequencing noise removal was used for variant selection and clonal analysis. Results: Complete concordance was seen for mutations detected in 23 of 27 patient-matched BM and PB paired samples. The likely cause of discordance in the other 4 pairs is attributable to mutations with low VAF ( < 0.5%). Overall, 91% of detected variants were concordant between PB and BM with strong correlation of VAFs (R = 0.9617). Serial testing trends correlated with stable disease with limited changes in VAFs (n = 2), and successful treatment response with rapidly falling VAFs (n = 5). In particular, 2 patients with FLT3-ITD and BCR-ABL1, respectively, identified at diagnosis had dramatic decrease in VAFs with respective targeted treatment. Conclusions: We report the utility of an ultrasensitive sequencing assay in PB with excellent concordance of mutations with BM, suggesting PB is a reliable alternative source for testing. In serial samples, the assay demonstrated value in treatment selection and response evaluation.

scholarly journals Seroprevalence of Anti-SARS-CoV-2 IgG and IgM among Adults over 65 Years Old in the South of Italy

Diagnostics ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 483
Author(s):  
Immacolata Polvere ◽  
Alfredina Parrella ◽  
Giovanna Casamassa ◽  
Silvia D’Andrea ◽  
Annamaria Tizzano ◽  
...  
Keyword(s):  
Antibody Response ◽  
Immunoglobulin M ◽  
Molecular Testing ◽  
Elisa Assay ◽  
The South ◽  
Serum Samples ◽  
Serological Screening ◽  
Rna Detection ◽  
In Vitro Diagnostic

SARS-CoV-2 is a zoonotic betacoronavirus associated with worldwide transmission of COVID-19 disease. By the beginning of March, WHO reported about 113,820,000 confirmed cases including more than 2,527,000 deaths all over the world. However, the true extent of virus circulation or its real infection/fatality ratio is not well-estimated due to the huge portion of asymptomatic infections. In this observational study, we have estimated the prevalence of specific immunoglobulin M and G directed towards SARS-CoV-2 antigen in a cohort of 1383 adult volunteers aged over 65 years old, living in the district of Benevento, in the South of Italy. Serological screening was carried out on capillary blood in September 2020, seven months after pandemic outbreak in Italy, to evaluate virus circulation and antibody response among elderly adults, in which severe symptoms due to viral infection are more common. The overall seroprevalence of anti-SARS-CoV-2 antibodies was 4.70% (CI 3.70%–5.95%) with no statistically significant differences between sexes. Among these, 69.69% (CI 55.61%–77.80%) tested positive to IgM, 23.08% (CI 14.51%–34.64%) to IgG and 9.23% (CI 4.30%–18.71%) was positive for both. All patients that were positive to IgM underwent molecular testing through RT-qPCR on oral-rhino pharyngeal swabs and only one specimen was positive for SARS-CoV-2 RNA detection. Instead, the presence of IgG from screened volunteers was confirmed by re-testing serum samples using both an ELISA assay validated for in vitro diagnostic use (IVD) and a recently published synthetic peptide-based ELISA assay. In conclusion, our report suggests that (1) early restrictions were successful in limiting COVID-19 diffusion in the district of Benevento; (2) rapid serological analysis is an ideal testing for both determining real seroprevalence and massive screening, whereas detection of viral RNA remains a gold standard for identification of infected patients; (3) even among people without COVID-19 related symptoms, the antibody response against SARS-CoV-2 antigens has individual features.

scholarly journals Evaluation of Microbial Culture Techniques for the Isolation of Pythium Insidiosum from Equine Tissues

2002 ◽  
Vol 14 (4) ◽  
pp. 288-294 ◽  
Author(s):  
Amy M. Grooters ◽  
Amy Whittington ◽  
Mae K. Lopez ◽  
Michelle N. Boroughs ◽  
Alma F. Roy
Keyword(s):  
Room Temperature ◽  
Microbial Culture ◽  
Sample Type ◽  
Pythium Insidiosum ◽  
Selective Media ◽  
Culture Techniques ◽  
Media Type ◽  
Antibiotic Solution ◽  
And Storage ◽  
Storage Technique

The purpose of this study was to evaluate the effects of sample handling, storage, and culture techniques on the isolation of Pythium insidiosum from infected equine tissues. Tissue and kunker samples obtained immediately posteuthanasia from a horse with subcutaneous pythiosis were used to assess the effects of sample type (kunkers vs. tissues), media type (selective vs. nonselective), storage technique, and storage time on P. insidiosum isolation rate. Overall, isolation rates were higher from fresh kunkers (94.6%) and stored kunkers (76.4%) than from fresh tissues (8.3%) or stored tissues (4.6%). Isolation of P. insidiosum also occurred more often on antibiotic-containing media than on nonselective media for both fresh and stored samples. For samples that were stored for 1–3 days prior to culture, P. insidiosum isolation rates were highest for the following techniques: kunkers stored at room temperature and plated on selective media (100%), kunkers stored at 4 C and then plated on either nonselective (91.7%) or selective (95.8%) media, kunkers stored on cold packs and then plated on either nonselective (93.8%) or selective (100%) media, kunkers stored in ampicillin solution and plated on selective media (100%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). For samples stored for 4–5 days, P. insidiosum isolation rates were highest for kunkers stored at 4 C and then plated on either nonselective (81.3%) or selective (87.5%) media, kunkers stored in ampicillin solution and then plated on selective media (87.5%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). Results of this study suggest that optimal isolation rates of P. insidiosum from infected equine tissues are achieved by culturing fresh kunkers on selective media. For samples that cannot be processed immediately, acceptable handling techniques include storage at room temperature for up to 3 days, refrigeration for up to 5 days, shipping on cold packs, and storage in antibiotic solution, each combined with subsequent inoculation on selective media.

scholarly journals STRESS AND DEFORMATION STUDY ON CASTELLATED STEEL BEAM WITH TAPERED SHAPE AND HEXAGONAL OPENINGS

SINERGI ◽  
2019 ◽  
Vol 23 (1) ◽  
pp. 61
Author(s):  
Taufiq Ilham Maulana ◽  
Bagus Soebandono ◽  
Aris Susanti
Keyword(s):  
Numerical Analysis ◽  
Open Source Software ◽  
Steel Beam ◽  
Sample Type ◽  
Span Length ◽  
3 Dimensional ◽  
Primary Sample ◽  
Cantilever Structure ◽  
Stress And Deformation ◽  
Regular Section

Castellated steel beam is a beam with a regular section cut into half with a particular pattern and regrouped with welding to increase its height compared to the original. This structure element has been developed in building constructions since many years ago. However, its uniform section along the span will make the modification no longer effective in cantilever structure, unless it has additional adaptation. Therefore, in this study, it is proposed to use a castellated steel beam with a tapered shape to be applied as cantilever structures. A steel beam with IWF section 150x75x5x7 is the primary sample type in this research. Some variations were made such as openings angle for 450 and 500, openings space for 50 mm, 70 mm, and 90 mm, openings diameter for 50 mm, 75 mm, and 100 mm, and span length for 2 m, 2.5 m, 3 m, and 3.5 m. Two open-source software namely FreeCAD and LisaFEA were used to draw solid 3-dimensional samples and to conduct the numerical analysis to determine stress and deformation respectively. From the result, it is known that the smallest stresses and deformations can be achieved by a different angle of openings, openings space, and diameter for each span length.

scholarly journals Extended storage of SARS-CoV-2 nasopharyngeal swabs does not negatively impact results of molecular-based testing across three clinical platforms

2020 ◽  
pp. jclinpath-2020-206738
Author(s):  
Karin A Skalina ◽  
D Y Goldstein ◽  
Jaffar Sulail ◽  
Eunkyu Hahm ◽  
Momka Narlieva ◽  
...  
Keyword(s):  
Real World ◽  
Nasopharyngeal Swab ◽  
Molecular Testing ◽  
Clinical Tests ◽  
Ambient Conditions ◽  
Long Term Stability ◽  
The Us ◽  
Us Fda ◽  
World Environment

With the global outbreak of COVID-19, the demand for testing rapidly increased and quickly exceeded the testing capacities of many laboratories. Clinical tests which receive CE (Conformité Européenne) and Food and Drug Administration (FDA) authorisations cannot always be tested thoroughly in a real-world environment. Here we demonstrate the long-term stability of nasopharyngeal swab specimens for SARS-CoV-2 molecular testing across three assays recently approved by the US FDA under Emergency Use Authorization. This study demonstrates that nasopharyngeal swab specimens can be stored under refrigeration or even ambient conditions for 21 days without clinically impacting the results of the real-time reverse transcriptase-PCR testing.

scholarly journals Confirmed or unconfirmed cases of 2019 novel coronavirus pneumonia in Italian patients: a retrospective analysis of clinical features

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Giulia De Angelis ◽  
Brunella Posteraro ◽  
Federico Biscetti ◽  
Gianluca Ianiro ◽  
Lorenzo Zileri Dal Verme ◽  
...  
Keyword(s):  
Clinical Features ◽  
Multivariable Analysis ◽  
Etiologic Agent ◽  
Nasopharyngeal Swab ◽  
Imaging Features ◽  
Rt Pcr ◽  
Rna Detection ◽  
Incubation Periods ◽  
Novel Coronavirus ◽  
To Receive

Abstract Background Since December 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a novel etiologic agent of viral pneumonia. We aimed to compare clinical features of 165 Italian patients with laboratory confirmed or unconfirmed 2019-nCoV pneumonia. Methods On March 31, 2020, hospitalized patients who presented with fever and/or respiratory symptoms, exposures, and presence of lung imaging features consistent with 2019-nCoV pneumonia were included. Before admission to a hospital ward, patients underwent RT-PCR based SARS-CoV-2 RNA detection in their nasopharyngeal swab samples. Results Of 165 patients studied, 119 had positive RT-PCR results and 46 were RT-PCR negative for 2 days or longer (i.e., when the last swab sample was obtained). The median age was 70 years (IQR, 58–78), and 123 (74.6%) of 165 patients had at least one comorbidity. The majority of patients (101/165, 61.2%) had a mild pneumonia, and the remaining patients (64/165, 38.8%) a severe/critical pneumonia. We did not find any substantial difference in symptoms, incubation periods, and radiographic/CT abnormalities as well as in many of the biological abnormalities recorded. However, at multivariable analysis, higher concentrations of hemoglobin (OR, 1.34; 95% CI, 1.11–1.65; P = 0.003) and lower counts of leukocytes (OR, 0.81; 95% CI, 0.72–0.90; P < 0.001) were statistically associated with confirmed COVID-19 diagnosis. While mortality rates were similar, patients with confirmed diagnosis were more likely to receive antivirals (95% vs 19.6%, P < 0.001) and to develop ARDS (63% vs 37%, P = 0.003) than those with unconfirmed COVID-19 diagnosis. Conclusions Our findings suggest that unconfirmed 2019-nCoV pneumonia cases may be actually COVID-19 cases and that clinicians should be cautious when managing patients with presentations compatible with COVID-19.

scholarly journals Red wine as resveratrol protective system -

2004 ◽  
Vol 22 (SI - Chem. Reactions in Foods V) ◽  
pp. S166-S168 ◽  
Author(s):  
I. Kolouchová ◽  
P. Zámostný ◽  
Z. Bělohlav ◽  
K. Melzoch ◽  
L. Siříšťová
Keyword(s):  
Room Temperature ◽  
Red Wine ◽  
Hplc Method ◽  
Isomerization Reaction ◽  
Sample Type ◽  
Red Wines ◽  
Liquid Sample ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Protective System

The study describes an isomerization of trans-resveratrol exposed to diffused daylight in dependence on temperature and character of liquid sample (type of solvent, wine …). These effects were investigated using the reverse-phase-HPLC method. Trans-resveratrol isoform was light sensitive and under daylight isomerized to cis-isoform very easily. The exposition of trans-resveratrol solution by higher temperatures did not induce isomerization reaction if that was kept in dark. The wines were analyzed immediately after opening (0 hour), and again after 48 hours and after 17 days. The concentration of resveratrol in tested red wines was stable at 4°C, under room temperature the total amount of resveratrol in all cases increased.

scholarly journals A large-volume sputum dry storage and transportation device for molecular and culture-based diagnosis of tuberculosis

2021 ◽  
Author(s):  
Andrea Dsouza ◽  
Saylee Jangam ◽  
Vishwanath Naik ◽  
J Manjula ◽  
Chandrasekhar B Nair ◽  
...  
Keyword(s):  
Glass Fiber ◽  
Large Volume ◽  
Liquid State ◽  
Mycobacterium Smegmatis ◽  
Room Temperature ◽  
Molecular Testing ◽  
Laboratory Services ◽  
State Recovery ◽  
Optimum Utilization ◽  
Sample Recovery

Technologies for preservation of specimens in the absence of cold chains are essential for optimum utilization of existing laboratory services in the developing world. We present a prototype called specimen transportation tube (SPECTRA-tube) for the collection, exposure-free drying, ambient transportation, and liquid state recovery of large-volume (>1 mL) specimens. Specimens introduced into SPECTRA-tube are dried in glass fiber membranes, which are critical for efficient liquid-state sample recovery by rehydration and centrifugation. Mycobacterium smegmatis (Msm)-spiked mock sputum dried in native Standard 17 glass fiber was stable for molecular testing after 10-day storage at 45°C, and for culture testing after 10- and 5-day storage at 37°C and 45°C, respectively. Compatibility with human sputum storage was demonstrated by dry storing Mycobacterium bovis-spiked pooled human sputum in SPECTRA-tube for 5 days at room temperature followed by successful qPCR detection. By significantly increasing the volume of samples that can be transported in the dry state and enabling recovery of the entire sample in liquid state, SPECTRA-tube presents a potential universal solution for the preservation and transportation of liquid specimens.

scholarly journals Molecular Detection of SARS-Cov-2 in Exhaled Breath Using a Portable Sampler

2021 ◽  
Author(s):  
Tim Stakenborg ◽  
Joren Raymenants ◽  
Ahmed Taher ◽  
Elisabeth Marchal ◽  
Bert Verbruggen ◽  
...  
Keyword(s):  
Temporal Dynamics ◽  
Respiratory Infections ◽  
Sample Volume ◽  
Nasopharyngeal Swab ◽  
Exhaled Breath ◽  
Sample Type ◽  
Viral Shedding ◽  
Current Test ◽  
Standard Sampling ◽  
Antigen Tests

Abstract The SARS-CoV-2 pandemic has highlighted the need for improved technologies to help control the spread of contagious pathogens. While rapid point-of-need testing plays a key role in strategies to rapidly identify and isolate infectious patients, a cornerstone for any disease-control strategy, current test approaches have significant shortcomings related to assay limitations and sample type. Direct quantification of viral shedding in exhaled particles may offer a better rapid testing approach, since SARS-CoV-2 is believed to spread mainly by aerosols. It potentially measures contagiousness directly, the sample is easy to obtain, its production can be standardized between patients, and the limited sample volume lends itself to a fast and sensitive analysis. In view of these benefits, we developed and tested an approach where exhaled particles are efficiently sampled using inertial impaction in a micromachined silicon chip, followed by an in-situ RT-qPCR molecular assay to detect SARS-CoV-2 shedding. We demonstrate that sampling subjects using a one-minute breathing protocol, yields sufficient viral RNA to detect infections with a sensitivity comparable to standard sampling methods. A longitudinal study revealed clear differences in the temporal dynamics of viral load for nasopharyngeal swab, saliva, breath, and antigen tests. Overall, after an infection, the breath-based test is the first to consistently report a negative result, putatively signaling the end of contagiousness and further emphasizing the potential of this tool to help manage the spread of airborne respiratory infections. 


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