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1.
Microbiol Spectr ; 11(3): e0006423, 2023 06 15.
Article in English | MEDLINE | ID: mdl-37097146

ABSTRACT

Appropriate interpretation of various diagnostic tests for COVID-19 is critical, yet the association among rapid antigen tests, reverse transcription (RT)-PCR, and viral culture has not been fully defined. To determine whether rapid antigen testing correlates with the presence and quantity of replication-competent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in ambulatory adults, 626 adult participants were enrolled in a cross-sectional diagnostic study. Each participant had two anterior nasal swabs obtained for rapid antigen and RT-PCR testing and SARS-CoV-2 viral culture. The primary outcomes were the presence and quantification of SARS-CoV-2 growth in VeroE6-ACE2-TMPRSS2 cells in asymptomatic and symptomatic ambulatory adults. In this cross-sectional study of 626 adult outpatients, the sensitivity of a single positive antigen test to identify replication-competent SARS-CoV-2 was 63.6% in asymptomatic and 91.0% in symptomatic participants. Viral culture titers were the highest at the onset of symptoms and rapidly declined by 7 days after symptom onset. The positive agreement of the rapid antigen test with RT-PCR at a cycle threshold CT less than 30 was 66.7% in asymptomatic and 90.7% in symptomatic participants. Among symptomatic participants a with a CT less than 30, a single antigen test had a positive agreement of 90.7% (95% confidence interval [CI], 84.8% to 94.8%). There was 100% negative agreement as all 425 RT-PCR-negative participants had a negative antigen test. A positive antigen test in symptomatic adults with COVID-19 has a strong correlation with replication-competent SARS-CoV-2. Rapid antigen test results may be a suitable proxy for infectiousness. IMPORTANCE Do rapid antigen test results correlate with replication-competent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (i.e., infectious) virus? In this cross-sectional diagnostic study of 626 adults, the sensitivity of the antigen test to identify replication-competent SARS-CoV-2 was 63.6% in asymptomatic and 91.0% in symptomatic participants. Viral culture titers were the highest at the onset of symptoms and rapidly declined by 7 days after symptom onset. The positive agreement of the rapid antigen test with reverse transcription (RT)-PCR at a CT of less than 30 was 66.7% in asymptomatic participants and 90.7% in symptomatic participants. A positive antigen test may be an appropriate surrogate for identifying replication-competent virus in symptomatic individuals with COVID-19.


Subject(s)
COVID-19 , SARS-CoV-2 , Adult , Humans , SARS-CoV-2/genetics , COVID-19/diagnosis , Cross-Sectional Studies , Polymerase Chain Reaction , Outpatients
3.
Lancet Reg Health Southeast Asia ; 3: 100036, 2022 Aug.
Article in English | MEDLINE | ID: mdl-35784831

ABSTRACT

Background: Additional outpatient therapies which are readily accessible will be essential to reduce COVID-19 illness progression in high risk individuals. Especially as the virus continues to mutate with greater transmissibility despite increased global vaccination. Methods: A randomized, double-blind, multicentre, parallel group, placebo-controlled phase III clinical trial evaluated the ability of nitric oxide (NO) to rapidly eradicate nasal SARS-CoV-2 RNA. Adults (18-70 years) with mild symptomatic COVID-19 were randomized, confirmed by laboratory SARS-CoV-2 reverse transcription polymerase chain reaction (RT-PCR) nasal swab. Randomisation was 1:1, NONS (N = 153) vs placebo (N = 153). NO generated by a nasal spray (NONS) was self-administered six times daily as two sprays per nostril (0⋅45 mL of solution/dose) for seven days. Patients at high risk of illness progression, defined as unvaccinated, ≥ 45 years of age or having comorbidities, were the primary analysis population. Findings: Overall, mean SARS-CoV-2 RNA concentrations (6·96 log10 copies/mL in the NONS group and 7·16 log10 copies/mL in the placebo group) were comparable at baseline. Primary endpoint mean treatment difference SARS-CoV-2 RNA change from baseline to the end of treatment (EOT) was -0·52 copies/mL (SE 0·202, 95% CI -0·92 to -0·12; p = 0·010) with NONS compared to placebo. Secondary endpoint assessments demonstrated a greater proportion of patients receiving NONS (82·8%) cleared SARS-CoV-2 (RT-PCR negative) by EOT compared to placebo (66·7%, p = 0·046), with no virus RNA detected a median of four days earlier compared to placebo (three vs seven days; p = 0·044). Interpretation: Use of NONS in patients recently infected with SARS-CoV-2 accelerates nasal virus clearance. Funding: Funding provided by Glenmark Pharmaceuticals Limited. Study medication provided by SaNOtize.

5.
Elife ; 102021 04 20.
Article in English | MEDLINE | ID: mdl-33876726

ABSTRACT

Diagnosis of SARS-CoV-2 (COVID-19) requires confirmation by reverse transcription-polymerase chain reaction (RT-PCR). Abbott ID NOW provides fast results but has been criticized for low sensitivity. Here we determine the sensitivity of ID NOW in an ambulatory population presented for testing. The study enrolled 785 symptomatic patients, of whom 21 were positive by both ID NOW and RT-PCR, and 2 only by RT-PCR. All 189 asymptomatic patients tested negative. The positive percent agreement between the ID NOW assay and the RT-PCR assay was 91.3%, and negative percent agreement was 100%. The results from the current study were included into a larger systematic review of literature where at least 20 subjects were simultaneously tested using ID NOW and RT-PCR. The overall sensitivity for ID NOW assay was calculated at 84% (95% confidence interval 55-96%) and had the highest correlation to RT-PCR at viral loads most likely to be associated with transmissible infections.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19/diagnosis , Reverse Transcriptase Polymerase Chain Reaction/methods , SARS-CoV-2/isolation & purification , Adult , Female , Genome, Viral , Humans , Male , Middle Aged , Nose/virology , Nucleic Acid Amplification Techniques/methods , Point-of-Care Systems , SARS-CoV-2/genetics , Sensitivity and Specificity , Specimen Handling , Viral Load
6.
PLoS One ; 16(4): e0245423, 2021.
Article in English | MEDLINE | ID: mdl-33852576

ABSTRACT

BACKGROUND: In order to identify an inexpensive yet highly stable SARS-CoV-2 collection device as an alternative to foam swabs stored in transport media, both contrived ("surrogate") CoV-positive and patient-collected spun polyester swabs stored in dry tubes were evaluated for time- and temperature-stability using qPCR. METHODS: Surrogate specimens were prepared by combining multiple, residual SARS-CoV-2-positive clinical specimens and diluting to near-LOD levels in either porcine or human mucus ("matrix"), inoculating foam or polyester nasal swabs, and sealing in dry tubes. Swabs were then subjected to one of three temperature excursions: (1) 4°C for up to 72 hours; (2) 40°C for 12 hours, followed by 32°C for up to 60 hours; or (3) multiple freeze-thaw cycles (-20°C). The stability of extracted SARS-CoV-2 RNA for each condition was evaluated by qPCR. Separate usability studies for the dry polyester swab-based HealthPulse@home COVID-19 Specimen Collection Kit were later conducted in both adult and pediatric populations. RESULTS: Polyester swabs stored dry demonstrated equivalent performance to foam swabs for detection of low and moderate SARS-CoV-2 viral loads. Mimicking warm- and cold- climate shipment, surrogate specimens were stable following either 72 hours of a high-temperature excursion or two freeze-thaw cycles. In addition, usability studies comprised of self-collected patient specimens yielded sufficient material for molecular testing, as demonstrated by RNase P detection. CONCLUSIONS: Polyester nasal swabs stored in dry collection tubes offer a robust and inexpensive self-collection method for SARS-CoV-2 viral load testing, as viral RNA remains stable under conditions required for home collection and shipment to the laboratory.


Subject(s)
COVID-19/diagnosis , COVID-19/virology , SARS-CoV-2/isolation & purification , Specimen Handling/methods , Animals , COVID-19 Testing/methods , Clinical Laboratory Techniques/methods , Diagnostic Tests, Routine/methods , Humans , Molecular Diagnostic Techniques , Nasopharynx/virology , Polyesters , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction/methods , SARS-CoV-2/genetics , Swine
7.
PLoS One ; 15(10): e0241100, 2020.
Article in English | MEDLINE | ID: mdl-33108384

ABSTRACT

Both polyester and foam nasal swabs were collected from convalescent COVID-19 patients at a single visit and stored in viral transport media (VTM), saline or dry. Sensitivity of each swab material and media combination were estimated, three by three tables were constructed to measure polyester and foam concordance, and cycle threshold (Ct) values were compared. 126 visits had polyester and foam swabs stored in viral transport media (VTM), 51 had swabs stored in saline, and 63 had a foam swab in VTM and a polyester swab stored in a dry tube. Polyester and foam swabs had an estimated sensitivity of 87.3% and 94.5% respectively in VTM, 87.5% and 93.8% respectively in saline, and 75.0% and 90.6% respectively for dry polyester and foam VTM. Polyester and foam Ct values were correlated, but polyester showed decreased performance for cases with a viral load near the detection threshold and higher Ct values on average.


Subject(s)
Betacoronavirus/isolation & purification , Clinical Laboratory Techniques , Convalescence , Coronavirus Infections/virology , Nasal Cavity/virology , Pandemics , Pneumonia, Viral/virology , Polyesters , Polyurethanes , Specimen Handling/instrumentation , Adult , Betacoronavirus/genetics , COVID-19 , COVID-19 Testing , Coronavirus Infections/diagnosis , Culture Media , Disposable Equipment/supply & distribution , Female , Health Personnel , Humans , Male , Middle Aged , Pneumonia, Viral/diagnosis , RNA, Viral/analysis , Random Allocation , Reverse Transcriptase Polymerase Chain Reaction , SARS-CoV-2 , Saline Solution , Sensitivity and Specificity , Specimen Handling/methods , Viral Load
8.
Crit Care Med ; 48(11): 1680-1689, 2020 11.
Article in English | MEDLINE | ID: mdl-32826428

ABSTRACT

OBJECTIVES: We explore ways to reduce errors in laboratory diagnosis of severe acute respiratory syndrome-coronavirus 2 infection by considering preanalytic, analytic, and postanalytic sources. To address preanalytic challenges, we first consider alternative anatomic sites for specimen collection, then discuss self-collection, alternative sampling devices, and transport media. Strengths and limitations of various analytic test systems are considered in the context of postanalytic challenges associated with making test results meaningful, specifically considering the complex relationship between "positive" test results and reproduction and shedding of intact virus. Finally, we provide recommendations regarding healthcare worker surveillance and release of patients with coronavirus disease 2019 from isolation. DATA SOURCES: Material was derived from a Webinar available to the public, manufacturer's websites, U.S. Food and Drug Administration, and Centers for Disease Control and Prevention websites and from both peer-reviewed papers identified by PubMed search and nonpeer-reviewed papers posted on Biorxiv and Medrxiv. Unpublished data came from the Washington State Department of Health. STUDY SELECTION: We included studies that compared diagnostic performance strategies without introducing bias due to use of an imperfect gold standard. Case series and case reports were included as necessary to illuminate the significance of results. DATA EXTRACTION: Data were extracted manually. DATA SYNTHESIS: Sensitivity, specificity, and CIs were computed from article data using a composite reference standard. Nucleic acid-based tests were assumed to perform at 100% specificity. CONCLUSIONS: Although sputum and bronchoalveolar lavage samples provide the highest diagnostic sensitivity for severe acute respiratory syndrome-coronavirus 2, nasopharyngeal, mid turbinate, and nasal specimens are suitable in most cases and require less use of personal protective equipment. When desired sampling materials are unavailable, alternatives may be substituted with no loss of performance. Both reverse transcriptase polymerase chain reaction tests and rapid nucleic acid-based tests offer good performance in most circumstances. Testing is not required to release most patients from isolation.


Subject(s)
Betacoronavirus/isolation & purification , Coronavirus Infections/diagnosis , Pneumonia, Viral/diagnosis , Specimen Handling/standards , COVID-19 , COVID-19 Testing , Centers for Disease Control and Prevention, U.S. , Clinical Laboratory Techniques , Humans , Pandemics , Reverse Transcriptase Polymerase Chain Reaction , SARS-CoV-2 , United States
10.
Sci Rep ; 9(1): 17872, 2019 Nov 25.
Article in English | MEDLINE | ID: mdl-31767913

ABSTRACT

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

11.
Sci Rep ; 9(1): 6427, 2019 04 23.
Article in English | MEDLINE | ID: mdl-31015537

ABSTRACT

We aimed to compare the efficacy and safety of Multipulse laser vaporesection of the prostate (MPVP) versus plasmakinetic resection of the prostate (PKRP) for treatment of patients with benign prostate obstruction (BPO) in a prospective trial. From January 2016 to April 2017, a total of 144 patients were included in the cohort study, of whom 73 patients underwent MPVP and 71 underwent PKRP. All patients received pre-operative evaluation and followed up at 1, 3, 6 and 12 months postoperatively. Baseline characteristics, perioperative data and postoperative outcomes were compared. Early (within 30 days postoperatively) and late complications were also recorded. Preoperative data, including age, prostate volume, international prostate symptom score (IPSS), International Index of Erectile Function Questionnaires (IIEF-5), the rate of anticoagulants use, Charlson comorbidity index were similar in two groups. Peri-operative parameters, including the rate of transfusion, and decrease in hemoglobin level were comparable. The operative time, the duration of catheterization and length of hospital stay were significantly shorter in the MPVP group. The voiding parameters and the quality-of-life scores (QoL) improved significantly in both groups postoperatively. There was a significantly difference in QoL at 1-year in the MPVP group (p < 0.001), under mixed model analysis with random effect and Bonferroni correction. There were no significant differences in improvement of IPSS, Qmax, IIEF-5, residual prostate volume ratio and PSA level reduction at the 1-year follow-up. MPVP was significantly superior to PKRP in terms of a reduction in overall complication rate (21.9% vs 45.0%, p = 0.004). Both treatments led to comparable symptomatic improvements. MPVP demonstrates satisfactory efficiency, shorter catheterization time and shorter hospital stay. Our data revealed that MPVP may be a promising technique which is safe and favorable alternative for patients with BPO.


Subject(s)
Laser Therapy/methods , Prostate/surgery , Prostatic Hyperplasia/surgery , Transurethral Resection of Prostate/methods , Aged , Dysuria/diagnosis , Dysuria/etiology , Dysuria/physiopathology , Hematuria/diagnosis , Hematuria/etiology , Hematuria/physiopathology , Humans , Laser Therapy/adverse effects , Lasers , Length of Stay/statistics & numerical data , Male , Middle Aged , Operative Time , Organ Size , Penile Erection/physiology , Postoperative Complications/diagnosis , Postoperative Complications/physiopathology , Prospective Studies , Prostate/pathology , Prostate/physiopathology , Prostatic Hyperplasia/pathology , Prostatic Hyperplasia/physiopathology , Surveys and Questionnaires , Transurethral Resection of Prostate/adverse effects , Treatment Outcome , Urethral Stricture/diagnosis , Urethral Stricture/etiology , Urethral Stricture/physiopathology , Urinary Incontinence, Urge/diagnosis , Urinary Incontinence, Urge/etiology , Urinary Incontinence, Urge/physiopathology , Urination/physiology
12.
Life Sci ; 92(1): 35-41, 2013 Jan 17.
Article in English | MEDLINE | ID: mdl-23142244

ABSTRACT

AIMS: Studies have shown that heme oxygenase-1 (HO-1) has a protective role in the mechanism underlying hypoxic preconditioning. We used a far-infrared radiation (FIR) heater to investigate the postconditioning protective role of HO-1 against ischemia/reperfusion (I/R) injury in rat testis. MAIN METHODS: Forty rats were used. Testis ischemia was mimicked by total obstructive clamping of testis vessels for 1, 2, or 4 h, and concomitant postconditioning with 30 min FIR or heat light during initially 30 min reperfusion. HO-1 expression and apoptosis of testis tissues were examined by immunohistochemistry and in situ terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay, respectively. HO-1 protein level and caspase-3 activity were analyzed by Western blotting. KEY FINDINGS: There was less apoptotic activity in rat testis after FIR, as determined by TUNEL assay. Higher HO-1 protein expression was observed by immunohistochemistry and Western blotting (p<0.01) in testis cells after FIR postconditioning. In contrast, caspase-3 activity was significantly higher in heat light groups, as compared with FIR groups (p<0.01). SIGNIFICANCE: FIR postconditioning attenuated I/R injury in rat testis by inducing HO-1 expression, which might have a protective role in testis apoptosis after I/R injury.


Subject(s)
Heme Oxygenase-1/genetics , Infrared Rays , Ischemic Postconditioning/methods , Reperfusion Injury/prevention & control , Testis/pathology , Animals , Apoptosis/radiation effects , Blotting, Western , Caspase 3/metabolism , Gene Expression Regulation/radiation effects , Hot Temperature , Immunohistochemistry , In Situ Nick-End Labeling , Light , Male , Rats , Rats, Wistar , Reperfusion Injury/pathology , Testis/blood supply , Time Factors
13.
J Photochem Photobiol B ; 106: 61-8, 2012 Jan 05.
Article in English | MEDLINE | ID: mdl-22062776

ABSTRACT

Far infrared (FIR) irradiation has been widely applied in health promotion. The aims of this study were to investigate the protective effect of FIR irradiation on stressed keratinocytes and the signaling pathways involved. HaCaT was subjected to sorbitol dehydration with or without 40min pretreatment with FIR radiation 4h earlier. Western blots of cell lysates were analyzed for caspase-3, HO-1, BCL2, Bax, ERK, and Akt. The incidence of apoptosis was also assessed by TUNEL staining. Evaluation of cell viability was determined using MTT. mRNAs were extracted and compared using Illumina Human Ref-8 v2 BeadChips. Hyperosomotic injury of HaCaT cells caused by sorbitol resulted in increased cleaved caspase-3 expression and this effect was decreased by FIR pretreatment; these findings were confirmed by TUNEL staining and MTT tests. Pre-treatment with FIR irradiation before sorbitol-induced dehydration significantly upregulated phosphorylated Akt (p-Akt) levels and A6730, an Akt kinase inhibitor (5µM), attenuated the protective effect of FIR irradiation. A microarray study showed FIR irradiation had far less effect at the transcriptional level. FIR pretreatment attenuates apoptosis and cell death in dehydration-stressed cultured keratinocytes through the PI-3K/Akt pathway, this protective effect of FIR irradiation is not at the transcriptional level.


Subject(s)
Apoptosis/radiation effects , Infrared Rays , Keratinocytes/radiation effects , Caspase 3/metabolism , Cell Line , Humans , Keratinocytes/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Phosphorylation , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Signal Transduction , Sorbitol/chemistry , bcl-2 Homologous Antagonist-Killer Protein/metabolism , bcl-2-Associated X Protein/metabolism
14.
Toxicol Lett ; 207(3): 242-50, 2011 Dec 15.
Article in English | MEDLINE | ID: mdl-21968084

ABSTRACT

We evaluate the protective role of simvastatin-induced HO-1 in remote preconditioning against testis ischemia-reperfusion (IR) injury in vivo. Simvastatin was intraperitoneally (i.p.) injected 24 h before IR injury. Testis was occluded in the right testis for 40 min and followed by 30 min of reperfusion to induce IR injury. Tin protoporphyrin (Snpp), a competitive inhibitor of hemeoxygenase, was i.p. injected 1 h before the IR injury in separate groups of rats. The rat testes were harvested 24 h later. Induction of HO-1 expression by simvastatin was significantly increased at 24 and 48 h. Rats pre-treated with simvastatin showed higher expression of HO-1 protein by Western blotting and immunohistochemistry (IHC), and presented lower caspases-3 activity by caspase-3 activity assay. TUNEL staining analysis revealed simvastatin pretreatment significantly reduced IR induced cellular apoptosis. Contrarily, the simvastatin-induced cytoprotective effect was entirely abolished by administrations of Snpp. Further, lower caspase-3 activities were also noted in simvastatin plus Snpp (SS) group than the control plus Snpp (CS) group. After IR injury, eNOS immunoreactivity was markedly increased in the germ cell and Leydig cell of testicular tissues. Pretreatment of simvastatin significantly decreased eNOS immunoreactivity in the germ cell of the tubules in the rat testes. In conclusion, we suggest HO-1 plays a protective role in IR-induced injury in the testes of rats.


Subject(s)
Heme Oxygenase-1/biosynthesis , Ischemia/drug therapy , Reperfusion Injury/drug therapy , Simvastatin/pharmacology , Testis/drug effects , Animals , Apoptosis/drug effects , Blotting, Western , Caspase 3/metabolism , Enzyme Induction/drug effects , Heme Oxygenase-1/antagonists & inhibitors , In Situ Nick-End Labeling , Injections, Intraperitoneal , Ischemia/enzymology , Male , Metalloporphyrins/pharmacology , Protoporphyrins/pharmacology , Rats , Rats, Wistar , Reperfusion Injury/enzymology , Simvastatin/administration & dosage , Testis/blood supply
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