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1.
Viruses ; 13(4)2021 04 10.
Artigo em Inglês | MEDLINE | ID: mdl-33920072

RESUMO

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) RNA is detected by reverse-transcription quantitative real-time PCR (RT-qPCR) from respiratory specimens. This study compares throat washings (TW), nasopharyngeal swabs (NS) and oropharyngeal swabs (OS). A total of 102 samples from 34 adult patients with confirmed SARS-CoV-2 infection were analysed by RT-qPCR with absolute quantification. The median concentrations and diagnostic sensitivities were 5.8×104 copies/mL, 85% (NS), 1.4×104, 79% (OS) and 4.3×103, 85% (TW). Concentration differences were significant between NS and TW (P = 0.019). Saliva (SA) was available from 21 patients (median 3.4×103). OS and TW can be considered for SARS-CoV-2 diagnostics, although with slightly lower concentrations.


Assuntos
/diagnóstico , Faringe/virologia , Manejo de Espécimes/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RNA Viral/análise , RNA Viral/genética , Saliva/virologia , Sensibilidade e Especificidade , Carga Viral , Adulto Jovem
2.
Infect Dis Poverty ; 10(1): 45, 2021 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-33789752

RESUMO

BACKGROUND: The management of discharge COVID-19 patients with recurrent positive SARS-CoV-2 RNA is challenging. However, there are fewer scientific dissertations about the risk of recurrent positive. The aim of this study was to explore the relationship between SARS-COV-2 RNA positive duration (SPD) and the risk of recurrent positive. METHODS: This case-control multi-center study enrolled participants from 8 Chinese hospital including 411 participants (recurrent positive 241). Using unadjusted and multivariate-adjusted logistic regression analyses, generalized additive model with a smooth curve fitting, we evaluated the associations between SPD and risk of recurrent positive. Besides, subgroup analyses were performed to explore the potential interactions. RESULTS: Among recurrent positive patients, there were 121 females (50.2%), median age was 50 years old [interquartile range (IQR): 38-63]. In non-adjusted model and adjusted model, SPD was associated with an increased risk of recurrent positive (fully-adjusted model: OR = 1.05, 95% CI: 1.02-1.08, P = 0.001); the curve fitting was not significant (P = 0.286). Comparing with SPD < 14 days, the risk of recurrent positive in SPD > 28 days was risen substantially (OR = 3.09, 95% CI: 1.44-6.63, P = 0.004). Interaction and stratified analyses showed greater effect estimates of SPD and risk of recurrent positive in the hypertension, low monocyte count and percentage patients (P for interaction = 0.008, 0.002, 0.036, respectively). CONCLUSION: SPD was associated with a higher risk of recurrent positive and especially SPD > 28 day had a two-fold increase in the relative risk of re-positive as compared with SPD < 14 day. What's more, the risk may be higher among those with hypertension and lower monocyte count or percentage.


Assuntos
/virologia , RNA Viral/isolamento & purificação , /isolamento & purificação , Adulto , /patologia , Estudos de Casos e Controles , Feminino , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Faringe/virologia , RNA Viral/genética , Recidiva , Fatores de Risco , Fatores de Tempo , Eliminação de Partículas Virais
3.
BMC Infect Dis ; 21(1): 318, 2021 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-33823800

RESUMO

BACKGROUND: Accurate diagnosis in patients with suspected coronavirus disease 2019 (COVID-19) is essential to guide treatment and limit spread of the virus. The combined nasal and throat swab is used widely, but its diagnostic performance is uncertain. METHODS: In a prospective, multi-centre, cohort study conducted in secondary and tertiary care hospitals in Scotland, we evaluated the combined nasal and throat swab with reverse transcriptase-polymerase chain reaction (RT-PCR) for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in consecutive patients admitted to hospital with suspected COVID-19. Diagnostic performance of the index and serial tests was evaluated for a primary outcome of confirmed or probable COVID-19, and a secondary outcome of confirmed COVID-19 on serial testing. The diagnosis was adjudicated by a panel, who recorded clinical, laboratory and radiological features blinded to the test results. RESULTS: We enrolled 1368 consecutive patients (median age 68 [interquartile range, IQR 53-80] years, 47% women) who underwent a total of 3822 tests (median 2 [IQR 1-3] tests per patient). The primary outcome occurred in 36% (496/1368), of whom 65% (323/496) and 35% (173/496) had confirmed and probable COVID-19, respectively. The index test was positive in 255/496 (51%) patients with the primary outcome, giving a sensitivity and specificity of 51.4% (95% confidence interval [CI] 48.8 to 54.1%) and 99.5% (95% CI 99.0 to 99.8%). Sensitivity increased in those undergoing 2, 3 or 4 tests to 60.1% (95% CI 56.7 to 63.4%), 68.3% (95% CI 64.0 to 72.3%) and 77.6% (95% CI 72.7 to 81.9%), respectively. The sensitivity of the index test was 78.9% (95% CI 74.4 to 83.2%) for the secondary outcome of confirmed COVID-19 on serial testing. CONCLUSIONS: In patients admitted to hospital, a single combined nasal and throat swab with RT-PCR for SARS-CoV-2 has excellent specificity, but limited diagnostic sensitivity for COVID-19. Diagnostic performance is significantly improved by repeated testing.


Assuntos
/diagnóstico , Nariz/virologia , Faringe/virologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Hospitalização , Hospitais , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Escócia , Sensibilidade e Especificidade
4.
Indian J Med Microbiol ; 39(1): 122-124, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33610243

RESUMO

The route of transmission of Novel SARS-CoV-2 virus is ambiguous. In this regard we planned a study to find out SARS-CoV-RNA shedding in various clinical samples of 9 COVID-19 positive patients. SARS-CoV-RNA was detected in nasal swab (NS), throat swab (TS) and faecal sample but was not detected in serum and urine samples. We also report that SARS-CoV-2-RNA persisted in faeces for >20 days. Persistence of faecal RNA might impose challenge in infection control and the disease may spread to household contacts if discharged. Perineal cleaning and hygiene may be advised at the time of vaginal delivery.


Assuntos
/epidemiologia , RNA Viral , Adolescente , Adulto , Criança , Pré-Escolar , Fezes/virologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Cavidade Nasal/virologia , Faringe/virologia , Fatores de Tempo , Carga Viral , Adulto Jovem
5.
Zool Res ; 42(2): 161-169, 2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33554485

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and coronavirus disease 2019 (COVID-19) continue to impact countries worldwide. At present, inadequate diagnosis and unreliable evaluation systems hinder the implementation and development of effective prevention and treatment strategies. Here, we conducted a horizontal and longitudinal study comparing the detection rates of SARS-CoV-2 nucleic acid in different types of samples collected from COVID-19 patients and SARS-CoV-2-infected monkeys. We also detected anti-SARS-CoV-2 antibodies in the above clinical and animal model samples to identify a reliable approach for the accurate diagnosis of SARS-CoV-2 infection. Results showed that, regardless of clinical symptoms, the highest detection levels of viral nucleic acid were found in sputum and tracheal brush samples, resulting in a high and stable diagnosis rate. Anti-SARS-CoV-2 immunoglobulin M (IgM) and G (IgG) antibodies were not detected in 6.90% of COVID-19 patients. Furthermore, integration of nucleic acid detection results from the various sample types did not improve the diagnosis rate. Moreover, dynamic changes in SARS-CoV-2 viral load were more obvious in sputum and tracheal brushes than in nasal and throat swabs. Thus, SARS-CoV-2 nucleic acid detection in sputum and tracheal brushes was the least affected by infection route, disease progression, and individual differences. Therefore, SARS-CoV-2 nucleic acid detection using lower respiratory tract samples alone is reliable for COVID-19 diagnosis and study.


Assuntos
/veterinária , /genética , Animais , Anticorpos Antivirais , Modelos Animais de Doenças , Haplorrinos , Humanos , Estudos Longitudinais , Faringe/virologia , Valor Preditivo dos Testes , Manejo de Espécimes , Escarro/virologia
6.
Respiration ; 100(2): 116-126, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33486496

RESUMO

BACKGROUND: There is still no clinical evidence available to support or to oppose corticosteroid treatment for coronavirus disease 2019 (COVID-19) pneumonia. OBJECTIVE: To investigate the efficacy and safety of corticosteroid given to the hospitalized patients with COVID-19 pneumonia. METHODS: This was a prospective, multicenter, single-blind, randomized control trial. Adult patients with COVID-19 pneumonia who were admitted to the general ward were randomly assigned to either receive methylprednisolone or not for 7 days. The primary end point was the incidence of clinical deterioration 14 days after randomization. RESULTS: We terminated this trial early because the number of patients with COVID-19 pneumonia in all the centers decreased in late March. Finally, a total of 86 COVID-19 patients underwent randomization. There was no difference of the incidence of clinical deterioration between the methylprednisolone group and control group (4.8 vs. 4.8%, p = 1.000). The duration of throat viral RNA detectability in the methylprednisolone group was 11 days (interquartile range, 6-16 days), which was significantly longer than that in the control group (8 days [2-12 days], p = 0.030). There were no significant differences between the 2 groups in other secondary outcomes. Mass cytometry discovered CD3+ T cells, CD8+ T cells, and NK cells in the methylprednisolone group which were significantly lower than those in the control group after randomization (p < 0.05). CONCLUSIONS: From this prematurely closed trial, we found that the short-term early use of corticosteroid could suppress the immune cells, which may prolong severe acute respiratory syndrome coronavirus 2 shedding in patients with COVID-19 pneumonia. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04273321.


Assuntos
/tratamento farmacológico , Glucocorticoides/uso terapêutico , Hospitalização , Metilprednisolona/uso terapêutico , Faringe/química , RNA Viral/isolamento & purificação , Eliminação de Partículas Virais , Adulto , Idoso , Antibacterianos/uso terapêutico , Antivirais/uso terapêutico , Complexo CD3 , Linfócitos T CD8-Positivos , /terapia , Progressão da Doença , Intervenção Médica Precoce , Oxigenação por Membrana Extracorpórea , Feminino , Humanos , Células Matadoras Naturais , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Oxigenoterapia , Quartos de Pacientes , Faringe/virologia , Modelos de Riscos Proporcionais , Respiração Artificial , Método Simples-Cego , Subpopulações de Linfócitos T , Linfócitos T , Fatores de Tempo , Resultado do Tratamento
7.
Am J Otolaryngol ; 42(2): 102896, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33445037

RESUMO

OBJECTIVES: This study aims to propose a novel and effective throat swab collection method for coronavirus disease 2019 (COVID-19). METHODS: The subjects were randomly divided into two groups. The subjects were asked to open their mouth to make "ah" sound (traditional method) or simulate yawn (improved method) for throat swab collection. The usage of tongue depressor, collection time, adverse reactions and subjective discomfort (VAS score) were compared. The collection time, comprehensive indicators of adverse reactions and VAS score were also compared among three collectors. RESULTS: The tongue depressor was less used in the improved group (χ2 = 40.186, P < 0.01). The average collection time of the traditional group was 5.44 ± 2.97 and that of the improved group was 4.00 ± 2.31 (P < 0.01). The subjects in the improved group had fewer and milder adverse reactions. The VAS score of subjects in the improved group was lower than that in the traditional group (P < 0.01). Among different collectors, the collection time, comprehensive indicators of adverse reactions and VAS were the same as the overall trend. CONCLUSION: Simulating yawn is a safer and faster throat swab collection method.


Assuntos
/diagnóstico , Faringe/virologia , Manejo de Espécimes/métodos , Bocejo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Distribuição Aleatória , Fatores de Tempo , Escala Visual Analógica , Adulto Jovem
8.
J Virol Methods ; 287: 114008, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33160015

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of the COVID-19 pandemic. Although other diagnostic methods have been introduced, detection of viral genes on oro- and nasopharyngeal swabs by reverse-transcription real time-PCR (rRT-PCR) assays is still the gold standard. Efficient viral RNA extraction is a prerequisite for downstream performance of rRT-PCR assays. Currently, several automatic methods that include RNA extraction are available. However, due to the growing demand, a shortage in kit supplies could be experienced in several labs. For these reasons, the use of different commercial or in-house protocols for RNA extraction may increase the possibility to analyze high number of samples. Herein, we compared the efficiency of RNA extraction of three different commercial kits and an in-house extraction protocol using synthetic ssRNA standards of SARS-CoV-2 as well as in oro-nasopharyngeal swabs from six COVID-19-positive patients. It was concluded that tested commercial kits can be used with some modifications for the detection of the SARS-CoV-2 genome by rRT-PCR approaches, although with some differences in RNA yields. Conversely, EXTRAzol reagent was the less efficient due to the phase separation principle at the basis of RNA extraction. Overall, this study offers alternative suitable methods to manually extract RNA that can be taken into account for SARS-CoV-2 detection.


Assuntos
/métodos , RNA Viral/isolamento & purificação , /isolamento & purificação , Testes Diagnósticos de Rotina , Genes Virais/genética , Humanos , Limite de Detecção , Faringe/virologia , RNA Viral/análise , RNA Viral/genética , Reprodutibilidade dos Testes , /genética
9.
Diagn Microbiol Infect Dis ; 99(1): 115206, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33045499

RESUMO

The diagnosis of coronavirus disease-19 (COVID-19) relies on the detection of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) RNA by real-time reverse-transcription polymerase chain reaction in respiratory samples. Rapid increase in the COVID-19 cases across the world requires fast and efficient testing as testing capacity is a bottleneck in diagnosis. In this context, pooling strategy can be opted for rapid testing in a cost-effective manner. In this study, the authors have optimized and compared the effect of pooling (5 and 10 samples) before and after nucleic acid extraction. It was concluded that there was no significant difference in the SARS CoV-2 RNA detection in the pools prepared at sample or RNA level. Even after pooling, 10-fold dilution was detectable with 3-cycle threshold value change in both type of pools when compared with individual samples. Hence, sample pool size of 10 can be used in low-prevalent areas, and testing capacity can be substantially increased.


Assuntos
/métodos , /isolamento & purificação , Manejo de Espécimes/métodos , /normas , Genes Virais/genética , Humanos , Índia/epidemiologia , Nasofaringe/virologia , Faringe/virologia , RNA Viral/genética , RNA Viral/isolamento & purificação , Sensibilidade e Especificidade , Manejo de Espécimes/normas , Centros de Atenção Terciária
10.
Talanta ; 224: 121726, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33379001

RESUMO

The outbreak of COVID-19 caused by a novel Coronavirus (termed SARS-CoV-2) has spread to over 210 countries around the world. Currently, reverse transcription quantitative qPCR (RT-qPCR) is used as the gold standard for diagnosis of SARS-CoV-2. However, the sensitivity of RT-qPCR assays of pharyngeal swab samples are reported to vary from 30% to 60%. More accurate and sensitive methods are urgently needed to support the quality assurance of the RT-qPCR or as an alternative diagnostic approach. A reverse transcription digital PCR (RT-dPCR) method was established and evaluated. To explore the feasibility of RT-dPCR in diagnostic of SARS-CoV-2, a total of 196 clinical pharyngeal swab samples from 103 suspected patients, 77 close contacts and 16 supposed convalescents were analyzed by RT-qPCR and then measured by the proposed RT-dPCR. For the 103 fever suspected patients, 19 (19/25) negative and 42 (42/49) equivocal tested by RT-qPCR were positive according to RT-dPCR. The sensitivity of SARS-CoV-2 detection was significantly improved from 28.2% by RT-qPCR to 87.4% by RT-dPCR. For 29 close contacts (confirmed by additional sample and clinical follow up), 16 (16/17) equivocal and 1 negative tested by RT-qPCR were positive according to RT-dPCR, which is implying that the RT-qPCR is missing a lot of asymptomatic patients. The overall sensitivity, specificity and diagnostic accuracy of RT-dPCR were 91%, 100% and 93%, respectively. RT-dPCR is highly accurate method and suitable for detection of pharyngeal swab samples from COVID-19 suspected patients and patients under isolation and observation who may not be exhibiting clinical symptoms.


Assuntos
/métodos , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , /genética , /genética , Humanos , Faringe/virologia , Fosfoproteínas/genética , Poliproteínas/genética , Proteínas Virais/genética
11.
J Clin Virol ; 134: 104712, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33338894

RESUMO

BACKGROUND: Currently, there are two rapid antigen detection (RAD) kits from the WHO Emergency Use List for detecting SARS-CoV-2. OBJECTIVE: The Panbio COVID-19 Ag Rapid Test Device was selected to evaluate the performance for detecting SARS-CoV-2. STUDY DESIGN: Analytical sensitivity for the detection of SARS-CoV-2 virus was determined by limit of detection (LOD) using RT-PCR as a reference method. Clinical sensitivity was evaluated by using respiratory specimens collected from confirmed COVID-19 patients. RESULTS: The LOD results showed that the RAD kit was 100 fold less sensitive than RT-PCR. Clinical sensitivity of the RAD kit was 68.6 % for detecting specimens from COVID-19 patients. CONCLUSIONS: The RAD kit evaluated in the present study shared similar performance with another kit from the WHO Emergency Use List, the Standard Q COVID-19 Ag. Understanding the clinical characteristics of RAD kits can guide us to decide different testing strategies in different settings.


Assuntos
Antígenos Virais/análise , Kit de Reagentes para Diagnóstico/normas , /imunologia , /patologia , /métodos , Reações Cruzadas , Hong Kong , Humanos , Limite de Detecção , Nasofaringe/virologia , Faringe/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Organização Mundial da Saúde
12.
Clin Lab ; 66(12)2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33337843

RESUMO

BACKGROUND: Coronavirus disease (COVID-19) has affected more than 100 countries worldwide and the discharge criteria of patients with COVID-19 vary across different countries. In China, patients with two negative respiratory viral RNA tests taken at least one day apart can be discharged with no further quarantine required. Currently, PCR testing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in fecal sample is not routinely performed. METHODS: We present a patient with COVID-19, whose respiratory swabs became negative but fecal sample remained positive for SARS-CoV-2 RNA. RESULTS: Stool sample collected on 27th of February was still positive for SARS-CoV-2 RNA, 24 days after the first negative respiratory swab. CONCLUSIONS: Based on the experience from the 2003 SARS epidemic, we recommend that fecal RNA testing of SARS-CoV-2 should be incorporated into the discharge criteria to minimize the risk of transmission from the gastrointestinal tract.


Assuntos
/virologia , Convalescença , Fezes/virologia , Alta do Paciente/normas , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , /isolamento & purificação , Adulto , Anticorpos Antivirais/sangue , Antivirais/uso terapêutico , Infecções Assintomáticas , /tratamento farmacológico , Quimioterapia Combinada , Reações Falso-Negativas , Feminino , Humanos , Nasofaringe/virologia , Faringe/virologia , Timalfasina/uso terapêutico
13.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi ; 55(11): 1032-1036, 2020 Nov 07.
Artigo em Chinês | MEDLINE | ID: mdl-33210882

RESUMO

Objective: To investigate the clinical characteristics of patients with coronavirus disease 2019 (COVID-19) with pharyngeal symptoms. Methods: From January 10 to May 15, 2020, clinical data of 1 228 patients with COVID-19 in Renmin Hospital of Wuhan University was collected (554 males and 674 females, with the range of age from 10 to 95 years old, the average age was 55.2 years old). The patients were divided into pharyngeal symptoms group (PS, 126 cases) and non-pharyngeal symptoms group (Non-PS, 1 102 cases) according to the presence or absence of pharyngeal symptoms such as pharyngalgia, pharyngeal dryness, pharyngeal itching, and pharyngeal foreign body sensation. The clinical data in terms of age, sex, medical history, duration of symptoms, treatment time, clinical classification, pulmonary imaging findings, whole blood cell count, serum hypersensitivity C-reactive protein, C-reactive protein, procalcitonin were statistically analyzed between the two groups. Chi-square, Fisher's exact test and Mann-Whitney U test were used for statistical analysis. Results: The most common pharyngeal symptoms were pharyngalgia (59.52%, 75/126), followed by foreign body sensation (23.02%, 29/126), pharyngeal dryness (8.73%, 11/126), and itching (8.73%, 11/126). The median age of the patients in the PS group was 51.50 years old, which was less than 57.50 years old in the non-PS group, showing a significant difference (P<0.05). The female cases accounted for 65.08% (82/126), which was higher than 53.72% (592/1 102) of the non-PS group (P<0.05). The incidence of bilateral lung inflammation confirmed by CT images was 73.81% (93/126), which was significantly lower than 83.48% (920/1 102) in the non-PS group (P<0.05). No significant differences were shown in the proportion of patients with clinical types, treatment days, duration of symptoms, white blood cell count, lymphocyte count, lymphocyte percentage, eosinophil count, eosinophil percentage, hypersensitive C-reactive protein, C-reactive protein, D-dimer, procalcitonin and other indicators (P>0.05). Conclusions: The incidence of pharyngeal symptoms in patients with COVID-19 is 10.26%. Most of these symptoms occur before or at the same time as the common symptoms of the disease. Therefore, patients with such symptoms may bring a greater risk of infection to otolaryngologist. According to the current clinical classification criteria, pharyngeal symptoms have no obvious correlation with the degree of the disease; but the presence of pharyngeal symptoms may suggest a milder clinical presentation and a better prognosis.


Assuntos
Infecções por Coronavirus/diagnóstico , Faringe/virologia , Pneumonia Viral/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Betacoronavirus , Criança , China , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Doenças Faríngeas/virologia , Faringe/fisiopatologia , Estudos Retrospectivos , Adulto Jovem
14.
BMJ Case Rep ; 13(10)2020 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-33130584

RESUMO

During the global pandemic of COVID-19 accurate diagnosis of the infection by demonstrating SARS-CoV-2 viral RNA by PCR in specimens is crucial for therapeutic and preventative interventions. There have been instances where nasal and throat swabs have been negative despite the patient having typical clinical and radiological findings compatible with the disease. We report a case of a man in his late 50s, brought to the hospital following a cardiac arrest and prolonged unsuccessful resuscitation. The history was typical for COVID-19 with fever for 10 days and worsening shortness of breath. His throat and nasal swabs (after death) were negative for SARS-CoV-2. A limited diagnostic autopsy was performed after 27 days, and lung swabs confirmed presence of SARS-CoV-2. This case highlights the importance of lung swabs when initial upper respiratory tract swabs are negative and proves that the virus can be detected from dead human tissue almost a month later.


Assuntos
Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , DNA Viral/análise , Pulmão/virologia , Parada Cardíaca Extra-Hospitalar/terapia , Faringe/virologia , Pneumonia Viral/diagnóstico , Autopsia , Reanimação Cardiopulmonar/métodos , Serviço Hospitalar de Emergência , Reações Falso-Negativas , Evolução Fatal , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Reação em Cadeia da Polimerase/métodos
15.
Viruses ; 12(10)2020 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-33092065

RESUMO

BACKGROUND: RT-PCR on nasopharyngeal (NPS)/oropharyngeal swabs is the gold standard for diagnosis of SARS-CoV-2 infection and viral load monitoring. Oral fluid (OF) is an alternate clinical sample, easy and safer to collect and could be useful for COVID-19 diagnosis, monitoring viral load and shedding. METHODS: Optimal assay conditions and analytical sensitivity were established for the commercial Simplexa™ COVID-19 Direct assay adapted to OF matrix. The assay was used to test 337 OF and NPS specimens collected in parallel from 164 hospitalized patients; 50 bronchoalveolar lavage (BAL) specimens from a subgroup of severe COVID-19 cases were also analysed. RESULTS: Using Simplexa™ COVID-19 Direct on OF matrix, 100% analytical detection down to 1 TCID50/mL (corresponding to 4 × 103 copies (cp)/mL) was observed. No crossreaction with other viruses transmitted through the respiratory toute was observed. Parallel testing of 337 OF and NPS samples showed highly concordant results (κ = 0.831; 95 % CI = 0.771-0.891), and high correlation of Ct values (r = 0.921; p < 0.0001). High concordance and elevated correlation was observed also between OF and BAL. Prolonged viral RNA shedding was observed up to 100 days from symptoms onset (DSO), with 32% and 29% positivity observed in OF and NPS samples, respectively, collected between 60 and 100 DSO. CONCLUSIONS: Simplexa™ COVID-19 Direct assays on OF have high sensitivity and specificity to detect SARS-CoV-2 RNA and provide an alternative to NPS for diagnosis and monitoring SARS-CoV-2 shedding.


Assuntos
Betacoronavirus/fisiologia , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/virologia , Pneumonia Viral/virologia , Eliminação de Partículas Virais/fisiologia , Adulto , Idoso , Betacoronavirus/genética , Líquidos Corporais/virologia , Infecções por Coronavirus/diagnóstico , Testes Diagnósticos de Rotina , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular/métodos , Pandemias , Faringe/virologia , Pneumonia Viral/diagnóstico , RNA Viral/análise , Sensibilidade e Especificidade , Manejo de Espécimes , Carga Viral
17.
Pediatr Infect Dis J ; 39(10): e315-e317, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32932332

RESUMO

In the current study, we report on 4 children with confirmed SARS-CoV-2 infection, of which 3 of them were asymptomatic. These patients had both pharyngeal swabs and anal swabs testing during hospital or after discharge. All the 4 children showed long-time positive viral RNA in the stool specimens after pharyngeal swabs turned negative during the follow-up stage, especially in the asymptomatic children. The positive RNA in stool specimens of asymptomatic children last for more than 54 days after admission or 30 days after discharge.


Assuntos
Infecções Assintomáticas , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/virologia , Fezes/virologia , Pneumonia Viral/virologia , Betacoronavirus/genética , Criança , Pré-Escolar , Infecções por Coronavirus/patologia , Feminino , Seguimentos , Humanos , Masculino , Pandemias , Faringe/virologia , Pneumonia Viral/patologia , RNA Viral/genética , RNA Viral/isolamento & purificação
18.
Immunotherapy ; 12(15): 1139-1148, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32933369

RESUMO

Aim: Evaluating the incidence and course of COVID-19 in cancer patients treated with immunotherapy. Patients & methods: We reported the influenza-like illness events with diagnosis of COVID-19 within the patient cohort enrolled in the prospective observational multicenter INVIDIa-2 study in the single center of Parma. Results: Among 53 patients, eight experienced influenza-like illness during the influenza season 2019/2020, and three of them had diagnosis of COVID-19. They were males, elderly, with cardiovascular disease. Radiological features of COVID-19 pneumonitis were found in all of three cases, although the pharyngeal swab resulted positive in only two. Two of these three patients died due to respiratory failure. Conclusion: Cancer patients are at high risk of severe events from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.


Assuntos
Infecções por Coronavirus/epidemiologia , Imunoterapia , Neoplasias/epidemiologia , Neoplasias/terapia , Pneumonia Viral/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antineoplásicos Imunológicos/uso terapêutico , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/patologia , Infecções por Coronavirus/fisiopatologia , Feminino , Humanos , Incidência , Itália/epidemiologia , Pulmão/diagnóstico por imagem , Pulmão/patologia , Masculino , Pessoa de Meia-Idade , Pandemias , Faringe/virologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/patologia , Pneumonia Viral/fisiopatologia , Estudos Prospectivos , Fatores de Risco , Tomografia Computadorizada por Raios X
19.
J Clin Lab Anal ; 34(10): e23554, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32977349

RESUMO

BACKGROUND: To compare the diagnostic efficacy between two different real-time reverse transcription polymerase chain reaction (RT-PCR) test kits for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid detection and provide references for laboratories. METHODS: Throat swab samples from 18 hospitalized patients were clinically diagnosed with coronavirus disease 2019 (COVID-19) and 100 hospitalized patients without COVID-19 were collected. SARS-CoV-2 nucleic acid was detected in throat swab samples with RT-PCR test kits from Sansure Biotech Inc (Hunan, China) and Shanghai BioGerm Medical Biotechnology Co., Ltd.(Shanghai, China). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and kappa value were analyzed, and three parallel tests were performed with three weakly positive samples. RESULTS: The sensitivity, specificity, PPV, NPV, and kappa value of the Sansure PCR kit were 0.833, 1.000, 1.000, 0.971, and 0.894, respectively, and the sensitivity, specificity, PPV, NPV, and kappa value of the BioGerm PCR kit were 0.944, 1.000, 1.000, 0.990, and 0.966, respectively. For the three parallel tests, the coefficient of variation value of the BioGerm PCR kit in all three samples was the smallest for both the ORF1ab and N gene. CONCLUSION: The detection efficacy of the BioGerm PCR kit for SARS-CoV-2 nucleic acid detection was relatively higher than that of the Sansure PCR kit.


Assuntos
Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real , Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Humanos , Pandemias , Faringe/virologia , Valor Preditivo dos Testes , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase em Tempo Real/normas
20.
ACS Sens ; 5(10): 3043-3048, 2020 10 23.
Artigo em Inglês | MEDLINE | ID: mdl-32989986

RESUMO

Mass testing is fundamental to face the pandemic caused by the coronavirus SARS-CoV-2 discovered at the end of 2019. To this aim, it is necessary to establish reliable, fast, and cheap tools to detect viral particles in biological material so to identify the people capable of spreading the infection. We demonstrate that a colorimetric biosensor based on gold nanoparticle (AuNP) interaction induced by SARS-CoV-2 lends itself as an outstanding tool for detecting viral particles in nasal and throat swabs. The extinction spectrum of a colloidal solution of multiple viral-target gold nanoparticles-AuNPs functionalized with antibodies targeting three surface proteins of SARS-CoV-2 (spike, envelope, and membrane)-is red-shifted in few minutes when mixed with a solution containing the viral particle. The optical density of the mixed solution measured at 560 nm was compared to the threshold cycle (Ct) of a real-time PCR (gold standard for detecting the presence of viruses) finding that the colorimetric method is able to detect very low viral load with a detection limit approaching that of the real-time PCR. Since the method is sensitive to the infecting viral particle rather than to its RNA, the achievements reported here open a new perspective not only in the context of the current and possible future pandemics, but also in microbiology, as the biosensor proves itself to be a powerful though simple tool for measuring the viral particle concentration.


Assuntos
Betacoronavirus/química , Colorimetria/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Mucosa Nasal/virologia , Faringe/virologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Técnicas Biossensoriais , Ouro , Humanos , Proteínas de Membrana/química , Nanopartículas Metálicas , Pandemias , Fotoquímica , Reação em Cadeia da Polimerase , Manejo de Espécimes , Glicoproteína da Espícula de Coronavírus/química , Níveis Máximos Permitidos , Proteínas do Envelope Viral/química
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