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1.
Biomed Res Int ; 2020: 1708267, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33029489

RESUMO

We aimed to summarize reliable medical evidence by the meta-analysis of all published retrospective studies that examined data based on the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by clinical symptoms, molecular (RT-PCR) diagnosis, and characteristic CT imaging features in pregnant women. The MEDLINE, PubMed, Scopus, ISI Web of Science, ClinicalKey, and CINAHL databases were used to select the studies. Then, 384 articles were received, including the studies until 01/May/2020. As a result of the full-text evaluation, 12 retrospective articles covering all the data related were selected. A total of 181 pregnant cases with SARS-CoV-2 infections were included in the meta-analysis within the scope of these articles. According to the results, the incidence of fever was 38.1% (95% CI: 14.2-65%) and cough was 22% (95% CI: 10.8-35.2%) among all clinical features of pregnant cases with SARS-CoV-2 infection. So, fever and cough are the most common symptoms in pregnant cases with SARS-CoV-2 infection, and 91.8% (95% CI: 76.7-99.9%) of RT-PCR results are positive. Moreover, abnormal CT incidence is 97.9% (95% CI: 94.2-99.9%) positive. No case was death. However, as this virus spreads globally, it should not be overlooked that the incidence will increase in pregnant women and maybe in the risky group. RT-PCR and CT can be used together in an accurate and safe diagnosis. In conclusion, these findings will provide important guidance for current studies regarding the clinical features and correct detection of SARS-CoV-2 infection in pregnant women, as well as whether it will create emergency tables that will require the use of a viral drug.


Assuntos
Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/complicações , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/complicações , Pneumonia Viral/diagnóstico , Complicações Infecciosas na Gravidez/diagnóstico , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Feminino , Humanos , Pandemias , Pneumonia Viral/epidemiologia , Gravidez , RNA Viral/análise , RNA Viral/genética , Estudos Retrospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tomografia Computadorizada por Raios X
2.
PLoS One ; 15(10): e0240081, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33007008

RESUMO

Given the global spread of coronavirus disease (COVID-19), strict discharge standard is of great significance for the prevention and control of the epidemic, thus, the purpose of this study is to formulate more strict and scientific discharge standards. A total of 845 patients with mild and general COVID-19 who were considered to be discharged from hospital were included in this study. The median time from the onset of COVID-19 to the occurrence of two consecutive negative nucleic acid tests of these patients was 21 days. 223 of the 845 patients were tested again after two consecutive negative nucleic acid tests and 17.49% of the patients were positive. Moreover, 82.51% (184 of 223) of these patients experienced negative results from three consecutive nucleic acid tests, the median time from the onset of COVID-19 to the occurrence of three consecutive negative nucleic acid tests was 23 days (range: 3-56 days), and 38 of which were further tested after three consecutive negative nucleic acid tests, while about 5.26% (2 of 38) patients showed positive nucleic acid test results. Thus, we suggested that the patient should be negative for at least 3 consecutive nucleic acid tests before discharge, and the test time should be no earlier than the 23rd day since the onset of the disease.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Alta do Paciente , Pneumonia Viral/diagnóstico , RNA Viral/análise , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , China/epidemiologia , Técnicas de Laboratório Clínico , Infecções por Coronavirus/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/epidemiologia , Estudos Retrospectivos , Adulto Jovem
3.
Acta Med Indones ; 52(3): 283-289, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33020339

RESUMO

Coronavirus disease 2019 (COVID-19) is a new infectious disease that spreads very rapidly and therefore, WHO has declared it as a global pandemic disease. The main clinical symptoms found in COVID-19 patients are cough and fever; however, in some cases, diarrhea can be one of the early symptoms. The present case report describes a patient who came with a complaint of diarrhea without fever and she was later confirmed to be positive for COVID-19 during hospitalization. The presence of unspecified initial symptoms calls for greater vigilance from health workers in establishing diagnosis patients with COVID-19.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/complicações , Diarreia/etiologia , Pandemias , Pneumonia Viral/complicações , RNA Viral/análise , Idoso , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Diarreia/diagnóstico , Feminino , Humanos , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Fatores de Tempo
4.
J Breath Res ; 14(4): 042003, 2020 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-33021206

RESUMO

Diagnosis of SARS-COV-2 infection (COVID-19) is currently based on detection of the viral RNA in nasopharyngeal swab samples by reverse transcription polymerase chain reaction (RT-PCR). However, sampling via nasopharyngeal swabs frequently provokes sneezing or coughing, which results in increased risk of the viral dissemination and environmental contamination. Furthermore, the sensitivity associated with the PCR tests s limited to 60%-70%, which is mainly attributable to technical deficiency in sampling. Given that the disease is transmitted via exhaled aerosol and droplets, and that the exhaled breath condensate (EBC) is the established modality for sampling exhaled aerosol, detection of the viral RNA in EBC is a promising approach for safe and efficient diagnosis of the disease. Subjects are those patients who are diagnosed with COVID-19 by positive nasopharyngeal swab PCR test and admitted to Saitama Medical Center, Japan. EBC samples will be collected using an R-tube® or R-tubeVent® device. Collected EBC samples will be introduced into a nucleic acid purifier. The purified nucleic acids will undergo amplification through RT-PCR for detection and quantification of SARS-COV-2 RNA. To date we have collected eight samples from seven subjects. Among them, two samples from two subjects tested positive for SARS-COV-2 RNA by the RT-PCR. Reflecting the second wave of COVID-19 prevalence in Japan, new admissions of COVID-19 patients to the Saitama Medical Center are increasing, and we are expecting to collect at least 50 EBC samples from 25 patients before the end of this year.


Assuntos
Testes Respiratórios/instrumentação , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Aerossóis/análise , Betacoronavirus , Técnicas de Laboratório Clínico , Tosse , Expiração , Humanos , Japão , Pandemias , RNA Viral/análise , Projetos de Pesquisa , Manejo de Espécimes , Carga Viral
5.
J Infect Dev Ctries ; 14(9): 977-981, 2020 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-33031084

RESUMO

INTRODUCTION: Current studies suggest that tears and conjunctival secretions may be an important transmission route in coronavirus disease 2019 (COVID-19). The study aims to evaluate the presence of severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) virus in tears and conjunctival secretion of patients with COVID-19. METHODOLOGY: A prospective interventional case series study was performed, and 32 patients with COVID-19 were selected at the Pamukkale University Hospital from 15 to 22 May 2020. The tear and conjunctival samples were collected by a conjunctival swab. Each specimen was sent to the laboratory for reverse transcription-polymerase chain reaction (RT-PCR) analyses. To avoid cross-infection, gloves and personal protective equipment were changed after collecting each sample. RESULTS: 32 patients (18 male, 14 female) with Covid-19 were included in this cross-sectional study. The average age of the patients was 52.81 ± 16.76 years. By the time of the first collection of conjunctival-tear samples, the mean time of the onset of complaints was 6.84 ± 6.81 (1-35) days. Tear-conjunctival samples from 5 patients (16%) without conjunctivitis yielded positive PCR results, 3 of whom had positive and 2 negative nasopharyngeal PCR results. CONCLUSIONS: Five of 32 patients (16 %) without conjunctivitis or any eye symptoms had viral RNA in their tear-conjunctival samples. The possibility of transmission via tears and conjunctival secretions should be recognized even in the absence of conjunctivitis or other ocular manifestations.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico/métodos , Túnica Conjuntiva/virologia , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Lágrimas/virologia , Adulto , Idoso , Betacoronavirus/genética , Infecções por Coronavirus/transmissão , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/transmissão , Estudos Prospectivos , RNA Viral/análise , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa
6.
BMC Infect Dis ; 20(1): 644, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32873230

RESUMO

BACKGROUND: To explore the clinical features and CT findings of clinically cured coronavirus disease 2019 (COVID-19) patients with viral RNA positive anal swab results after discharge. METHODS: Forty-two patients with COVID-19 who were admitted to Yongzhou Central Hospital, Hunan, China, between January 20, 2020, and March 2, 2020, were tested for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using anal swab viral RT-PCR. In this report, we present the clinical characteristics and chest CT features of six patients with positive anal swab results and compare the clinical, laboratory, and CT findings between the positive and negative groups. RESULTS: The anal swab positivity rate for SARS-CoV-2 RNA in discharged patients was 14.3% (6/42). All six patients were male. In the positive group, 40% of the patients (2/5) had a positive stool occult blood test (OBT), but none had diarrhea. The median duration of fever and major symptoms (except fever) in the positive patients was shorter than that of the negative patients (1 day vs. 6 days, 4.5 days vs. 10.5 days, respectively). The incidence of asymptomatic cases in the positive group (33.3%) was also higher than that of the negative group (5.6%). There were no significant differences in the CT manifestation or evolution of the pulmonary lesions between the two groups. CONCLUSION: In our case series, patients with viral RNA positive anal swabs did not exhibit gastrointestinal symptoms, and their main symptoms disappeared early. They had similar CT features to the negative patients, which may be easier to be ignored. A positive OBT may indicate gastrointestinal damage caused by SARS-CoV-2 infection.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico por imagem , Alta do Paciente/estatística & dados numéricos , Pneumonia Viral/diagnóstico por imagem , RNA Viral/análise , Síndrome Respiratória Aguda Grave/diagnóstico por imagem , Adolescente , Adulto , Idoso , Canal Anal/virologia , Betacoronavirus/genética , Criança , Pré-Escolar , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Febre , Hospitalização , Hospitais , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Síndrome Respiratória Aguda Grave/epidemiologia , Síndrome Respiratória Aguda Grave/virologia , Tomografia Computadorizada por Raios X , Adulto Jovem
7.
J Orthop Trauma ; 34(10): e377-e381, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32947588

RESUMO

BACKGROUND: Hospitals worldwide have postponed all nonessential surgery during the COVID-19 pandemic, but non-COVID-19 patients are still in urgent need of care. Uncertainty about a patient's COVID-19 status risks infecting health care workers and non-COVID-19 inpatients. We evaluated the use of quantitative reverse transcription polymerase chain reaction (RT-qPCR) screening for COVID-19 on admission for all patients with fractures. METHODS: We conducted a retrospective cohort study of patients older than 18 years admitted with low-energy fractures who were tested by RT-qPCR for SARS-CoV-2 at any time during hospitalization. Two periods based on the applied testing protocol were defined. During the first period, patients were only tested because of epidemiological criteria or clinical suspicion based on fever, respiratory symptoms, or radiological findings. In the second period, all patients admitted for fracture treatment were screened by RT-qPCR. RESULTS: We identified 15 patients in the first period and 42 in the second. In total, 9 (15.8%) patients without clinical or radiological findings tested positive at any moment. Five (33.3%) patients tested positive postoperatively in the first period and 3 (7.1%) in the second period (P = 0.02). For clinically unsuspected patients, postoperative positive detection went from 3 of 15 (20%) during the first period to 2 of 42 (4.8%) in the second (P = 0.11). Clinical symptoms demonstrated high specificity (92.1%) but poor sensitivity (52.6%) for infection detection. CONCLUSIONS: Symptom-based screening for COVID-19 has shown to be specific but not sensitive. Negative clinical symptoms do not rule out infection. Protocols and separated areas are necessary to treat infected patients. RT-qPCR testing on admission helps minimize the risk of nosocomial and occupational infection. LEVEL OF EVIDENCE: Diagnostic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/epidemiologia , Pandemias , Pneumonia Viral/epidemiologia , RNA Viral/análise , Triagem/métodos , Ferimentos e Lesões/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Feminino , Seguimentos , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Pneumonia Viral/diagnóstico , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Espanha/epidemiologia , Ferimentos e Lesões/complicações , Ferimentos e Lesões/epidemiologia
9.
Ann Saudi Med ; 40(5): 373-381, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32954791

RESUMO

BACKGROUND: The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) has prompted a need for mass testing to identify patients with viral infection. The high demand has created a global bottleneck in testing capacity, which prompted us to modify available resources to extract viral RNA and perform reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to detect SARS-COV-2. OBJECTIVES: Report on the use of a DNA extraction kit, after modifications, to extract viral RNA that could then be detected using an FDA-approved SARS-COV-2 RT-qPCR assay. MATERIALS AND METHODS: Initially, automated RNA extraction was performed using a modified DNA kit on samples from control subjects, a bacteriophage, and an RNA virus. We then verified the automated extraction using the modified kit to detect in-lab propagated SARSCOV-2 titrations using an FDA approved commercial kit (S, N, and ORF1b genes) and an in-house primer-probe based assay (E, RdRp2 and RdRp4 genes). RESULTS: Automated RNA extraction on serial dilutions SARS-COV-2 achieved successful one-step RT-qPCR detection down to 60 copies using the commercial kit assay and less than 30 copies using the in-house primer-probe assay. Moreover, RT-qPCR detection was successful after automated RNA extraction using this modified protocol on 12 patient samples of SARS-COV-2 collected by nasopharyngeal swabs and stored in viral transport media. CONCLUSIONS: We demonstrated the capacity of a modified DNA extraction kit for automated viral RNA extraction and detection using a platform that is suitable for mass testing. LIMITATIONS: Small patient sample size. CONFLICT OF INTEREST: None.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Nasofaringe/virologia , Pneumonia Viral/diagnóstico , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Automação , Chlorocebus aethiops , Técnicas de Laboratório Clínico , Vírus da Encefalomiocardite/genética , Humanos , Levivirus/genética , Proteínas do Nucleocapsídeo/genética , Pandemias , RNA Replicase/genética , RNA Viral/análise , Análise de Sequência de RNA , Glicoproteína da Espícula de Coronavírus/genética , Células Vero , Proteínas do Envelope Viral/genética , Proteínas não Estruturais Virais/genética
10.
BMC Infect Dis ; 20(1): 670, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32933492

RESUMO

BACKGROUND: The 2014/15 Ebola outbreak in West Africa resulted in 11,000 deaths and massive strain on local health systems, and the ongoing outbreak in Democratic Republic of Congo has afflicted more than 3000 people. Accurate, rapid Ebola diagnostics suitable for field deployment would enable prompt identification and effective response to future outbreaks, yet remain largely unavailable. The purpose of this study was to assess the accuracy of three novel rapid diagnostic tests (RDTs): an Ebola, an Ebola-Malaria, and a Fever Panel test that includes Ebola, all from a single manufacturer. METHODS: We evaluated the three RDTs in 109 Ebola-positive and 96 Ebola-negative stored serum samples collected during the outbreak in Guinea in 2014/15, and tested by real-time polymerase chain reaction (RT-PCR). Sensitivity, specificity, and overall percent agreement were calculated for each RDT using RT-PCR as a reference standard, stratified by Ct value ranges. RESULTS: All tests performed with high accuracy on samples with low Ct value (high viral load). The Fever Panel test performed with the highest accuracy, with a sensitivity of 89.9% and specificity of 90.6%. The Ebola and Ebola-Malaria tests performed comparably to each other: sensitivity was 77.1 and 78% respectively, and specificity was 91.7% for the Ebola test and 95.8% for the Ebola-Malaria test. CONCLUSIONS: This study evaluated the accuracy of three novel rapid diagnostic tests for Ebola. The tests may have significant public health relevance, particularly the Fever Panel test, which detects seven pathogens including Ebola. Given limitations to the study resulting from uncertain sample quality, further evaluation is warranted. All tests performed with highest accuracy on samples with low Ct value (high viral load), and the data presented here suggests that these RDTs may be useful for point-of-care diagnosis of cases in the context of an outbreak. Restrictions to their use in non-severe Ebola cases or for longitudinal monitoring, when viral loads are lower, may be appropriate. Highlighting the challenge in developing and evaluating Ebola RDTs, there were concerns regarding sample integrity and reference testing, and there is a need for additional research to validate these assays.


Assuntos
Doença pelo Vírus Ebola/diagnóstico , Sistemas Automatizados de Assistência Junto ao Leito , Surtos de Doenças , Ebolavirus/genética , Ebolavirus/isolamento & purificação , Guiné/epidemiologia , Doença pelo Vírus Ebola/epidemiologia , Doença pelo Vírus Ebola/virologia , Humanos , RNA Viral/análise , RNA Viral/metabolismo , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade
11.
Curr Opin HIV AIDS ; 15(6): 345-350, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32969974

RESUMO

PURPOSE OF REVIEW: To discuss the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection by nucleic acid and antibody testing as well as its contribution to mitigating the spread of coronavirus disease 2019. RECENT FINDINGS: Nucleic acid testing (NAT) is the firstly developed and most widely used diagnostic technique for SARS-CoV-2 infection. However, the sensitivity of SARS-CoV-2 RNA NAT assays is always unsatisfactory, mainly due to insufficient viral RNA in samples, especially when upper respiratory samples were used. Compared with NAT assays, serological tests are more convenient and less dependent on the quality of sample collection. But the sensitivity of antibody assays varies largely to test samples collected at different time after onset of symptoms. The diagnostic sensitivity can be significantly improved by combination of RNA and antibody testing. Due to the lack of effective drugs and vaccines, population prevention results mainly from timely triage and quarantine of SARS-CoV-2 infected individuals. Thus, extensive testing with NAT and antibody assays simultaneously is very important to constrain coronavirus disease 2019 epidemic. SUMMARY: Viral RNA testing combining with serological testing could improve the early diagnosis of SARS-CoV-2 infection, which has great value for clinical practice and public health.


Assuntos
Betacoronavirus , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/prevenção & controle , Pandemias/prevenção & controle , Pneumonia Viral/diagnóstico , Pneumonia Viral/prevenção & controle , Técnicas de Laboratório Clínico , Diagnóstico Precoce , Humanos , RNA Viral/análise , Testes Sorológicos
12.
Acta Biomed ; 91(3): e2020003, 2020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32921701

RESUMO

BACKGROUND: The COVID-19 outbreak is now a pandemic disease reaching as much as 210 countries worldwide with more than 2.5 million infected people and nearly 200.000 deaths. Amplification of viral RNA by RT-PCR represents the gold standard for confirmation of infection, yet it showed false-negative rates as large as 15-20% which may jeopardize the effect of the restrictive measures taken by governments. We previously showed that several hematological parameters were significantly different between COVID-19 positive and negative patients. Among them aspartate aminotransferase and lactate dehydrogenase had predictive values as large as 90%. Thus a combination of RT-PCR and blood tests could reduce the false-negative rate of the genetic test. METHODS: We retrospectively analyzed 24 patients showing multiple and inconsistent RT-PCR, test during their first hospitalization period, and compared the genetic tests results with their AST and LDH levels. RESULTS: We showed that when considering the hematological parameters, the RT-PCR false-negative rates were reduced by almost 4-fold. CONCLUSIONS: The study represents a preliminary work aiming at the development of strategies that, by combining RT-PCR tests with routine blood tests, will lower or even abolish the rate of RT-PCR false-negative results and thus will identify, with high accuracy, patients infected by COVID-19.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Pandemias , Pneumonia Viral/diagnóstico , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Aspartato Aminotransferases/sangue , Biomarcadores/sangue , Infecções por Coronavirus/sangue , Infecções por Coronavirus/epidemiologia , Diagnóstico Diferencial , Reações Falso-Negativas , Feminino , Seguimentos , Testes Hematológicos/métodos , Humanos , Itália/epidemiologia , L-Lactato Desidrogenase/sangue , Masculino , Pessoa de Meia-Idade , Pneumonia Viral/sangue , Pneumonia Viral/epidemiologia , Reprodutibilidade dos Testes , Estudos Retrospectivos
13.
Acta Biomed ; 91(3): e2020014, 2020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32921710

RESUMO

BACKGROUND AND AIM: Isolation of subjects with active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a pivotal preventive measure in the ongoing coronavirus disease 2019 (COVID-19) pandemic. A growing number of studies reported cases of recurrent SARS-CoV-2 RNA positivity following disease recovery, which were identified with a critical literature search and then meta-analyzed in this article. MATERIALS AND METHODS: A digital search was performed in Medline and Web of Science, using the keywords "coronavirus disease 2019" OR "COVID-19" OR "severe acute respiratory disease 2" OR "SARS-CoV-2" AND "recurrence" OR "repositivization" OR "retesting", without date or language restrictions. Recovery was defined as resolution of symptoms, with at least two consecutive negative molecular tests. RESULTS: A total number of 17 studies, with 5,182 COVID-19 patients, were included. SARS-CoV-2 recurrent RNA positivity in recovered COVID-19 patients ranged between 7-23% across the studies, with follow-up testing between 1-60 days. The estimated cumulative rate of SARS-CoV-2 recurrent RNA positivity was 12% (95% confidence interval, 12-13%; I2, 74%). CONCLUSIONS: Repeated molecular testing on respiratory tracts specimens at 1 and 2 months after recovery from COVID-19 is strongly advisable for early identification, isolation and clinical management of subjects with SARS-CoV-2 recurrent RNA positivity.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/epidemiologia , Pandemias , Pneumonia Viral/epidemiologia , RNA Viral/análise , Recuperação de Função Fisiológica , Infecções por Coronavirus/virologia , Humanos , Pneumonia Viral/virologia
14.
Acta Biomed ; 91(3): e2020019, 2020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32921716

RESUMO

As the COVID 19 pandemic develops across the globe, a large amount of literature has been written about the different ways in which we can diagnose and investigate someone suspected of being infected with the new coronavirus. Many approaches highlight the importance of using reverse transcriptase polymerase chain reaction (RT-PCR) used in conjunction with computed tomography (CT) scans. Whilst CT scans have been shown to be useful, there are multiple risks associated with them, for example radiation exposure and the transmission risk associated with repeated use of a CT suite. Therefore, it is important to analyse their diagnostic ability and limitations and to consider other methods of diagnosing COVID 19. Additionally, RT-PCR testing can have significant rates of false negatives, indicating the importance of taking a more comprehensive diagnostic approach. Here, we aim to review and analyse this literature to compare RT-PCR, serum inflammatory biomarkers, chest radiographs, ultrasound and chest CT scanning as methods of diagnosing COVID 19, particularly in asymptomatic patients.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Pandemias , Pneumonia Viral/diagnóstico , RNA Viral/análise , Tomografia Computadorizada por Raios X/métodos , Biomarcadores/sangue , Infecções por Coronavirus/sangue , Infecções por Coronavirus/epidemiologia , Humanos , Pneumonia Viral/sangue , Pneumonia Viral/epidemiologia
15.
Acta Biomed ; 91(3): e2020025, 2020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32921721

RESUMO

BACKGROUND AND AIM OF THE WORK: The ongoing pandemic has elicited an increasing interest regarding the SARS-CoV-2 viral RNA detection in saliva specimens rather than through nasopharyngeal swabs. Our aim was to conduct a meta-analysis on the sensitivity and specificity of SARS-CoV-2 viral RNA detection through RT-qPCR based on salivary specimens compared to conventional nasopharyngeal swabs. METHODS: We reported our meta-analysis according to the PRISMA statement. We searched Pubmed, Embase, and pre-print archive medRxiv.og for eligible studies published up to June 1st, 2020. Raw data included true/false positive and negative tests, and the total number of tests. Sensitivity and specificity data were calculated for every study, and then pooled in a random-effects model. Heterogeneity was assessed using the I2 measure. Reporting bias was assessed by means of funnel plots and regression analysis. RESULTS: The systematic review eventually retrieved 14 studies including a total of 15 estimates, the were included in quantitative synthesis. We found a pooled specificity of 97.7% (95%CI 93.8-99.2) and a pooled sensitivity of 83.4% (95%CI 73.1-90.4), with an overall agreement assessed by means of Cohen's kappa equals to 0.750, 95%CI 0.62-0.88 (i.e. moderate agreement), with high heterogeneity and risk of reporting bias. CONCLUSIONS: In conclusion, diagnostic tests based on salivary specimens are somewhat reliable, but relatively few studies have been carried out. Moreover, such studies are characterized by low numbers and low sample power. Therefore, the of salivary samples is currently questionable for clinical purposes and cannot substitute other more conventional RT-qPCR based on nasopharyngeal swabs.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Nasofaringe/virologia , Pandemias , Pneumonia Viral/diagnóstico , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Saliva/virologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Humanos , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia
16.
Acta Biomed ; 91(3): e2020030, 2020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32921725

RESUMO

The coronavirus disease 2019 (COVID-19) pandemic is still spreading worldwide, affecting several million people. Unlike the previous two coronavirus outbreaks, COVID-19 has caused several thousand deaths for respiratory and multiple organ failure. As specifically concerns this latest infectious pathology, laboratory medicine can provide a substantial contribution to diagnosing an acute severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection through molecular testing, establishing the presence and extent of an immune response against the virus, mostly through serological testing. However, it can also help to predict the risk of unfavorable disease progression by measuring some conventional laboratory tests and, last but not least, can provide reliable therapeutic guidance. This article is hence aimed at offering recent updates on the important role and value of laboratory investigations in COVID-19, also providing information on some hot topics such as virus RNA detection in different biological samples, causes of recurrent positivity of reverse-transcription polymerase chain reaction (RT-PCR), potential strategies for enhancing the throughput of molecular testing (i.e., pre-test probability assessment, sample pooling, use of rapid tests), as well as pragmatic indications for enhancing the quality and value of serological testing and laboratory-based monitoring.


Assuntos
Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , RNA Viral/análise , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Humanos , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia
18.
Nat Commun ; 11(1): 4464, 2020 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-32900994

RESUMO

The SARS-CoV-2 pandemic has shown how a rapid rise in demand for patient and community sample testing can quickly overwhelm testing capability globally. With most diagnostic infrastructure dependent on specialized instruments, their exclusive reagent supplies quickly become bottlenecks, creating an urgent need for approaches to boost testing capacity. We address this challenge by refocusing the London Biofoundry onto the development of alternative testing pipelines. Here, we present a reagent-agnostic automated SARS-CoV-2 testing platform that can be quickly deployed and scaled. Using an in-house-generated, open-source, MS2-virus-like particle (VLP) SARS-CoV-2 standard, we validate RNA extraction and RT-qPCR workflows as well as two detection assays based on CRISPR-Cas13a and RT-loop-mediated isothermal amplification (RT-LAMP). In collaboration with an NHS diagnostic testing lab, we report the performance of the overall workflow and detection of SARS-CoV-2 in patient samples using RT-qPCR, CRISPR-Cas13a, and RT-LAMP. The validated RNA extraction and RT-qPCR platform has been installed in NHS diagnostic labs, increasing testing capacity by 1000 samples per day.


Assuntos
Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Betacoronavirus/isolamento & purificação , Bioensaio , Sistemas CRISPR-Cas , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/normas , Humanos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Pandemias , RNA Viral/análise , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade
19.
Nat Biotechnol ; 38(10): 1164-1167, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32948856

RESUMO

We measured severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentrations in primary sewage sludge in the New Haven, Connecticut, USA, metropolitan area during the Coronavirus Disease 2019 (COVID-19) outbreak in Spring 2020. SARS-CoV-2 RNA was detected throughout the more than 10-week study and, when adjusted for time lags, tracked the rise and fall of cases seen in SARS-CoV-2 clinical test results and local COVID-19 hospital admissions. Relative to these indicators, SARS-CoV-2 RNA concentrations in sludge were 0-2 d ahead of SARS-CoV-2 positive test results by date of specimen collection, 0-2 d ahead of the percentage of positive tests by date of specimen collection, 1-4 d ahead of local hospital admissions and 6-8 d ahead of SARS-CoV-2 positive test results by reporting date. Our data show the utility of viral RNA monitoring in municipal wastewater for SARS-CoV-2 infection surveillance at a population-wide level. In communities facing a delay between specimen collection and the reporting of test results, immediate wastewater results can provide considerable advance notice of infection dynamics.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Pandemias , Pneumonia Viral/epidemiologia , RNA Viral/análise , Águas Residuárias/virologia , Vigilância Epidemiológica Baseada em Águas Residuárias , Betacoronavirus/genética , Biotecnologia , Connecticut/epidemiologia , Humanos , Prevalência , RNA Viral/genética , Esgotos/virologia , Fatores de Tempo
20.
Elife ; 92020 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-32894217

RESUMO

Understanding and mitigating SARS-CoV-2 transmission hinges on antibody and viral RNA data that inform exposure and shedding, but extensive variation in assays, study group demographics and laboratory protocols across published studies confounds inference of true biological patterns. Our meta-analysis leverages 3214 datapoints from 516 individuals in 21 studies to reveal that seroconversion of both IgG and IgM occurs around 12 days post-symptom onset (range 1-40), with extensive individual variation that is not significantly associated with disease severity. IgG and IgM detection probabilities increase from roughly 10% at symptom onset to 98-100% by day 22, after which IgM wanes while IgG remains reliably detectable. RNA detection probability decreases from roughly 90% to zero by day 30, and is highest in feces and lower respiratory tract samples. Our findings provide a coherent evidence base for interpreting clinical diagnostics, and for the mathematical models and serological surveys that underpin public health policies.


Assuntos
Betacoronavirus/genética , Betacoronavirus/imunologia , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , RNA Viral/análise , Anticorpos Antivirais/sangue , Anticorpos Antivirais/isolamento & purificação , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/sangue , Infecções por Coronavirus/diagnóstico , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/isolamento & purificação , Imunoglobulina M/isolamento & purificação , Pandemias , Pneumonia Viral/sangue , Pneumonia Viral/diagnóstico , RNA Viral/isolamento & purificação
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