scholarly journals Implementation of a Rapid RT-LAMP Saliva-based SARS-CoV-2 Testing Program in the Workplace

2022 ◽  
Author(s):  
Bradley W.M. Cook ◽  
Kaitlyn Kobasa ◽  
Marielou Tamayo ◽  
Natasha Theriault ◽  
Diane J.R. Gordon Pappas ◽  
...  
Keyword(s):  
Lamp Assay ◽  
Rapid Testing ◽  
Routine Testing ◽  
Non Invasive ◽  
Qrt Pcr ◽  
Household Contacts ◽  
Human Sample ◽  
Two Samples ◽  
Saliva Collection ◽  
Second Wave

Rising SARS-CoV-2 cases, testing delays and the risk of pre-symptomatic and asymptomatic transmission provided the impetus for an in-house rapid testing pro-gram. Employees and their household contacts were encouraged to self-collect saliva samples which were pooled for routine testing using an established colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. In brief, individual or a maximum of four saliva samples were pooled, heat-inactivated to render microorganisms, especially SARS-CoV-2, non-infectious prior to being added to RT-LAMP assay tubes containing either human sample control gene, RNase P or a region of the SARS-CoV-2 gene, ORF1ab. During the second wave of SARS-CoV-2 infections in November 2020, two samples from an employee and a member of their household tested positive via RT-LAMP within two days of each other. A delayed clinical qRT-PCR test confirmation of both individuals 5 days later underscores the power of routine rapid testing with within-the-hour turnaround times. Workplace rapid testing programs using RT-LAMP are flexible in their design, have a reduced cost compared to qRT-PCR, may involve non-invasive self-saliva collection for increased safety for the testing personnel, and can be performed with minimal training.

scholarly journals Single-Sided Portable NMR Investigation to Assess and Monitor Cleaning Action of PVA-Borax Hydrogel in Travertine and Lecce Stone

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3697
Author(s):  
Valeria Stagno ◽  
Chiara Genova ◽  
Nicole Zoratto ◽  
Gabriele Favero ◽  
Silvia Capuani
Keyword(s):  
Pore Size ◽  
Experimental Conditions ◽  
Non Invasive ◽  
Bedding Planes ◽  
Paramagnetic Impurities ◽  
Two Samples ◽  
Action Time ◽  
Paramagnetic Agents ◽  
Portable Nmr ◽  
Non Destructive

In this work, we investigated the potential of PVA-borax hydrogel for cleaning limestones and the dependence of the cleaning on the porosity of the rock and on the action time of the hydrogel treatment. Towards this goal, we used a nuclear magnetic resonance (NMR) spectrometer, developed for non-invasive and non-destructive applications on cultural heritage. T2-NMR parameters were quantified on different samples of Lecce stone and Travertine cut perpendicular (Pe) and parallel (Pa) to the bedding planes under different experimental conditions: untreated samples, treated with Paraloid B72 and cleaned with PVA-PEO-borax hydrogel applied for 4 min and 2 h. The T2 results suggest that the effectiveness of the cleaning strongly depended on the porosity of the stones. In Lecce stone, the hydrogel seemed to eliminate both the paramagnetic impurities (in equal measure with 4 min and 2 h treatment) and Paraloid B72. In Travertine Pe, characterized by a smaller pore size compared to Lecce stone, no significant effects were found regarding both the cleaning and the treatment with Paraloid B72. In Travertine Pa, characterized by a larger pore size than the other two samples, the hydrogel seemed to clean the paramagnetic agents (it worked better if applied for a longer time) but it did not appear to have any effect on Paraloid B72 removal.

scholarly journals A Large Cohort Study of SARS-CoV-2 Detection in Saliva: A Non-Invasive Alternative Diagnostic Test for Patients with Bleeding Disorders

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2361
Author(s):  
Josiane Iole França Lopes ◽  
Carlos Alexandre da Costa Silva ◽  
Rodrigo Guimarães Cunha ◽  
Alexandra Martins Soares ◽  
Maria Esther Duarte Lopes ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Hospitalized Patients ◽  
Bleeding Disorders ◽  
Hematological Diseases ◽  
Non Invasive ◽  
Qrt Pcr ◽  
Large Cohort Study ◽  
The Mean ◽  
Sensitivity Specificity ◽  
Higher Sensitivity

Diagnosis of SARS-CoV-2 infections is mostly based on the nasopharyngeal swabs (NPS). However, this collection is invasive and uncomfortable, especially for children and patients with coagulopathies, whose NPS collection often causes bleeding. Thus, the aim of this study was to evaluate the usefulness and accuracy of saliva for the diagnosis of COVID-19 in patients presenting bleeding disorders. Samples of NPS, oropharyngeal swabs (OPS), and saliva were collected simultaneously from 1159 hospitalized patients with hematological diseases and from 524 healthcare workers, both symptomatic and asymptomatic for SARS-CoV-2. All samples were evaluated for SARS-CoV-2 by qRT-PCR. SARS-CoV-2 was detected in NPS, OPS and saliva from 16.9%, 14.4% and 15.6% individuals, respectively. Tests in saliva showed sensitivity, specificity, and overall agreement of 73.3%, 96.9% and 92.7% (=0.74), respectively. Salivary tests had good accuracy (AUC = 0.7) for discriminating negative and positive qRT-PCR for SARS-CoV-2. Higher sensitivity was observed in symptomatic than in non-symptomatic patients, as well as in healthy subjects than in patients with hematological disease, in both OPS and saliva. The mean viral load in NPS was significantly higher than in OPS and in saliva samples (p < 0.001). Saliva is a good diagnostic tool to detect SARS-CoV-2, especially among patients symptomatic for COVID-19, and is a valuable specimen for mass screening of hospitalized patients with hematological diseases, especially for those that with bleeding disorders.

scholarly journals Serum lncRNA TINCR Serve as a Novel Biomarker for Predicting the Prognosis in Triple-Negative Breast Cancer

2020 ◽  
Vol 19 ◽  
pp. 153303382096557
Author(s):  
Xiaojie Wang ◽  
Shuang Li ◽  
Huiyu Xiao ◽  
Xiaoqin Deng
Keyword(s):  
Breast Cancer ◽  
Benign Breast Disease ◽  
Triple Negative ◽  
Human Cancer ◽  
Breast Disease ◽  
Clinicopathological Features ◽  
Protein Coding ◽  
Non Invasive ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr

Background: Tissue differentiation-inducing non-protein coding RNA (TINCR) has been shown to play a crucial role in pathogenesis of various types of human cancer including breast cancer (BC). The purpose of this study was to determine the potential prognostic value of serum lncRNA TINCR in BC. Methods: Quantitative reverse transcription PCR (qRT-PCR) was performed to detect serum lncRNA TINCR levels in 72 triple-negative BC (TNBC) patients, 105 non-TNBC patients, 60 benign breast disease patients and 86 healthy subjects. Results: The results showed that serum lncRNA TINCR level was significantly increased in BC, especially in TNBC. High circulating lncRNA TINCR was significantly correlated with worse clinicopathological features and clinical outcome of TNBC. Multivariate analysis revealed that serum lncRNA TINCR was an independent prognostic factor for overall survival of TNBC. However, little association was found between serum lncRNA TINCR and the prognosis of non-TNBC. Conclusions: Taken together, our findings demonstrate that serum lncRNA TINCR might be a useful novel and non-invasive biomarker for the prognosis prediction of TNBC.

scholarly journals COVID-19 salivary signature: diagnostic and research opportunities

2020 ◽  
pp. jclinpath-2020-206834 ◽  
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.

Expression Profile Analysis and Target Gene Prediction of Three Conserved MicroRNAs in Resistant Gossypium hirsutum Cv. KV-1 Responding to Verticillium dahliae

2014 ◽  
Vol 884-885 ◽  
pp. 441-445
Author(s):  
Xiao Hong He ◽  
Min Shi ◽  
Quan Sun ◽  
Ying Fan Cai
Keyword(s):  
Verticillium Wilt ◽  
Deep Sequencing ◽  
Verticillium Dahliae ◽  
Upland Cotton ◽  
Target Genes ◽  
Gene Prediction ◽  
Interaction Mechanism ◽  
Sequencing Data ◽  
Qrt Pcr ◽  
Two Samples

Plant microRNAs (miRNAs) play important roles in the post-transcriptional regulation of plant growth, development, flowering, metabolism, and responses to stress. Verticillium wilt is a vascular disease in plants caused by the fungal pathogen Verticillium dahliae. In order to find and investigate miRNAs related to the upland cotton variety Zhongzhimian KV-1 resistant Verticillium wilt, deep sequencing technology was used to construct small RNA libraries of two samples, which from seedlings of KV-1 cotton by different pathogenicity strains Verticillium wilt pathogen infections. The V. dahliae strains D07038 and V991 were used in this study and are moderately virulent and virulent, respectively. miRNAs with differential expression among the samples were obtained through analysis of sequencing data and three miRNAs (miR1423a-5p, miR3444a-5p and miR5562) were chosen to be identified by quantitative real-time RT-PCR (qRT-PCR). At the same time, their target genes were predicted. The results of qRT-PCR were consistent, which indicated 3444a-5p and miR5562 were with the highest expression level in virulent condition, but miR1423a-5p was a low-level expression. The results of experiments agreed with deep sequencing data basically. Analysis of the transcript data for target genes of three conserved miRNAs indicated that they play an important role in plant-pathogen interaction mechanism. The identification and characterisation of miRNAs from upland cotton may help to further the study of miRNA regulatory mechanisms that are involved in resistance to Verticillium wilt.

scholarly journals Salivary proteomics in monitoring the therapeutic response of canine oral melanoma

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0256167
Author(s):  
Sekkarin Ploypetch ◽  
Sittiruk Roytrakul ◽  
Janthima Jaresitthikunchai ◽  
Narumon Phaonakrop ◽  
Patharakrit Teewasutrakul ◽  
...  
Keyword(s):  
Therapeutic Response ◽  
Cost Effective ◽  
Term Survival ◽  
Non Invasive ◽  
Oral Melanoma ◽  
Liquid Chromatography Tandem Mass ◽  
Novel Biomarkers ◽  
Saliva Collection ◽  
Short Term Survival

Saliva biomarkers are suitable for monitoring the therapeutic response of canine oral melanoma (COM), because saliva directly contacts the tumor, and saliva collection is non-invasive, convenient and cost effective. The present study aimed to investigate novel biomarkers from the salivary proteome of COM treated with surgery and a chemotherapy drug, carboplatin, 1–6 times, using a liquid chromatography–tandem mass spectrometry approach. The expression of a potential salivary biomarker, ubiquitin D (UBD), was observed and verified by western blot analysis. A significantly increased ratio of free UBD (fUBD) to conjugated UBD (cUBD) was shown in the pre-surgery stage (PreS) in OM dogs with short-term survival (STS) (less than 12 months after surgery) compared with that with long-term survival (more than 12 months after surgery). In dogs with STS, the ratio was also shown to be augmented in PreS compared with that after surgery, followed by treatment with carboplatin twice, 4 and 5 times [After treatment (AT)2, AT4 and AT5]. In addition, the expression of fUBD was enhanced in PreS compared with that of AT2 in the STS group. In conclusion, this study revealed that a ratio of fUBD to cUBD in PreS was plausibly shown to be a potential prognostic biomarker for survival in dogs with OM.

scholarly journals SARS-CoV-2 infections in kindergartens and associated households at the start of the second wave in Berlin, Germany – a cross sectional study

2020 ◽  
Author(s):  
Marlene Thielecke ◽  
Stefanie Theuring ◽  
Welmoed van Loon ◽  
Franziska Hommes ◽  
Marcus A. Mall ◽  
...  
Keyword(s):  
Signs And Symptoms ◽  
Cross Sectional Study ◽  
Kindergarten Children ◽  
Sectional Study ◽  
Cross Sectional ◽  
Routine Testing ◽  
Household Members ◽  
Precautionary Measures ◽  
Second Wave ◽  
Pandemic Wave

AbstractObjectivesThe comparatively large proportion of asymptomatic SARS-CoV-2 infections in the youngest children opens up the possibility that kindergartens represent reservoirs of infection. However, actual surveys in kindergartens beyond individual outbreaks are rare. At the beginning of the second pandemic wave in Berlin, Germany, i.e., end of September 2020, we screened SARS-CoV-2 infections among kindergarten children, staff and connected household members.MethodsTwelve kindergartens were randomly selected in the Berlin metropolitan area, and a total of 720 participants were recruited (155 pre-school children, 78 staff, 487 household members). Participants were briefly examined and interviewed, and SARS-CoV-2 infections and anti-SARS-Cov-2 IgG antibodies were assessed.ResultsSigns and symptoms, largely resembling common cold, were present in 24.2% of children and 28.9% of staff. However, no SARS-CoV-2 infection was detected among 701 PCR-tested individuals, and only one childcare worker showed IgG seroreactivity (0.15%; 1/672).ConclusionsAgainst a backdrop of increased pandemic activity in the community, this cross-sectional study does not suggest that kindergartens are silent transmission reservoirs. Nevertheless, at increasing pandemic activity, reinforced precautionary measures and repeated routine testing appears advisable.

scholarly journals tRNA-Derived Fragments in Seminal Plasma Exosomes Are Non-invasive Biomarkers for Diagnosis of Non-obstructive Azoospermia With Spermatogenic Failure

2020 ◽  
Author(s):  
Zuobin Zhu ◽  
Xiaoxiao Han ◽  
Ying Li ◽  
Yao Zhu ◽  
Zhenbei Li ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Predictive Accuracy ◽  
Preoperative Evaluation ◽  
Bioinformatic Analysis ◽  
Testicular Tissue ◽  
Obstructive Azoospermia ◽  
Diagnostic Biomarker ◽  
Testicular Sperm ◽  
Non Invasive ◽  
Qrt Pcr

Abstract BackgroundThere is lack of accurate and non-invasive preoperative evaluation for improving microdissection testicular sperm success rate. tRNA-derived small RNA (tsRNAs) perform a variety of physiological functions and are related to many physiological and pathological processes. This study sought to identify the potential of exosomal tsRNA as a novel non-invasive diagnostic biomarker for non-obstructive azoospermia (NOA) with spermatogenic failure.MethodsSeminal plasma exosome tsRNA levels were used in a two-stage (screened by tsRNA sequencing on Illumina NextSeq instrument and validated by qRT-PCR) case-control designed study. The expression levels of the selected tsRNAs were further examined in the testicular tissues of NOA patients and obstructive azoospermia (OA) patients, and their underlying role in the pathogenesis of non-obstructive azoospermia was performed by bioinformatic analysis.ResultsIn this study, two tsRNAs (tRF-Val-AAC-010: AUC = 0.96, specificity = 80%, sensitivity = 95%; tRF-Pro-AGG-003: AUC = 0.96, specificity = 87%, sensitivity=95%) were found to have a good predictive accuracy which also showed differential expression in testicular tissue between NOA patients and OA patients. We also found that the combinations of tRF-Val-AAC-010 and tRF-pro -AGG-003 have a better discriminating ability between NOA patients and OA patients than single biomarkers. Finally, the bioinformatic analysis showed that the two selected tsRNAs were involved in spermatogenesis.ConclusionThis study first evaluated tsRNAs as potential biomarkers for NOA diagnosis and identified the exosomal tRF-Val-AAC-010 and tRF-pro-AGG-003 in seminal plasma valuable as biomarkers for NOA diagnosis.

scholarly journals A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages

2020 ◽  
Author(s):  
Boon-Teong Teoh ◽  
Kim-Ling Chin ◽  
Nur-Izyan Samsudin ◽  
Shih-Keng Loong ◽  
Sing-Sin Sam ◽  
...  
Keyword(s):  
Detection Limit ◽  
Sensitivity And Specificity ◽  
Reverse Transcription ◽  
Zika Virus ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Zikv Infection ◽  
Qrt Pcr

Abstract Background: Early detection of Zika virus (ZIKV) infection during the viremia and viruria facilitates proper patient management and mosquito control measurement to prevent disease spread. Therefore, a cost-effective nucleic acid detection method for the diagnosis of ZIKV infection, especially in resource-deficient settings, is highly required.Methods: In the present study, a single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of both the Asian and African-lineage ZIKV. The detection limit, strain coverage and cross-reactivity of the ZIKV RT-LAMP assay was evaluated. The sensitivity and specificity of the RT-LAMP were also evaluated using a total of 24 simulated clinical samples. The ZIKV quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used as the reference assay.Results: The detection limit of the RT-LAMP assay was 3.73 ZIKV RNA copies (probit analysis, P ≤ 0.05). The RT-LAMP assay detected the ZIKV genomes of both the Asian and African lineages without cross-reacting with other arthropod-borne viruses. The sensitivity and specificity of the RT-LAMP assay were 90% (95% CI = 59.6 – 98.2) and 100% (95% CI = 78.5 – 100.0), respectively. The RT-LAMP assay detected ZIKV genome in 9 of 24 (37.5%) of the simulated clinical samples compared to 10 of 24 (41.7%) by qRT-PCR assay with a high level of concordance (k = 0.913, P < 0.001).Conclusion: The RT-LAMP assay is applicable for the broad coverage detection of both the Asian and African ZIKV strains in resource-deficient settings.

scholarly journals A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages

2020 ◽  
Author(s):  
Boon-Teong Teoh ◽  
Kim-Ling Chin ◽  
Nur-Izyan Samsudin ◽  
Shih-Keng Loong ◽  
Sing-Sin Sam ◽  
...  
Keyword(s):  
Detection Limit ◽  
Sensitivity And Specificity ◽  
Reverse Transcription ◽  
Zika Virus ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Zikv Infection ◽  
Qrt Pcr

Abstract Background: Early detection of Zika virus (ZIKV) infection during the viremia and viruria facilitates proper patient management and mosquito control measurement to prevent disease spread. Therefore, a cost-effective nucleic acid detection method for the diagnosis of ZIKV infection, especially in resource-deficient settings, is highly required.Methods: In the present study, a single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of both the Asian and African-lineage ZIKV. The detection limit, strain coverage and cross-reactivity of the ZIKV RT-LAMP assay was evaluated. The sensitivity and specificity of the RT-LAMP were also evaluated using a total of 24 simulated clinical samples. The ZIKV quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used as the reference assay.Results: The detection limit of the RT-LAMP assay was 3.73 ZIKV RNA copies (probit analysis, P ≤ 0.05). The RT-LAMP assay detected the ZIKV genomes of both the Asian and African lineages without cross-reacting with other arthropod-borne viruses. The sensitivity and specificity of the RT-LAMP assay were 90% (95% CI = 59.6 – 98.2) and 100% (95% CI = 78.5 – 100.0), respectively. The RT-LAMP assay detected ZIKV genome in 9 of 24 (37.5%) of the simulated clinical samples compared to 10 of 24 (41.7%) by qRT-PCR assay with a high level of concordance (k = 0.913, P < 0.001).Conclusion: The RT-LAMP assay is applicable for the broad coverage detection of both the Asian and African ZIKV strains in resource-deficient settings.

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