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1.
Euro Surveill ; 25(24)2020 06.
Artículo en Inglés | MEDLINE | ID: mdl-32583765

RESUMEN

Containment strategies and clinical management of coronavirus disease (COVID-19) patients during the current pandemic depend on reliable diagnostic PCR assays for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we compare 11 different RT-PCR test systems used in seven diagnostic laboratories in Germany in March 2020. While most assays performed well, we identified detection problems in a commonly used assay that may have resulted in false-negative test results during the first weeks of the pandemic.


Asunto(s)
Betacoronavirus/genética , Técnicas de Laboratorio Clínico/métodos , Infecciones por Coronavirus/diagnóstico , Equipo para Diagnóstico , Neumonía Viral/diagnóstico , Técnicas de Laboratorio Clínico/instrumentación , Heces/virología , Alemania , Humanos , Laboratorios , Reacción en Cadena de la Polimerasa Multiplex/instrumentación , Reacción en Cadena de la Polimerasa Multiplex/métodos , Pandemias , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/instrumentación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y Especificidad
2.
J Chin Med Assoc ; 83(6): 524-526, 2020 06.
Artículo en Inglés | MEDLINE | ID: mdl-32502116

RESUMEN

The rapid spread of coronavirus disease 2019 (COVID-19) in many countries causes citizens of daily inconvenience and even life-threat for elderly population. The invasion of the main pathogen, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; 2019 novel coronavirus [2019-nCoV]), into human body causes different levels of impact to various patients. One of the most important issues for COVID-19 is how to defend this virus with the ability to foresee the infected targets. Thus, we maintain the quarantined essentially as for as others saved from COVID-19. So far, the routine laboratory test to confirm whether infected by SARS-CoV-2/2019-nCoV or not is through real-time reverse transcription polymerase chain reaction (rRT-PCR; quantitative polymerase chain reaction [qPCR]) with certain sequence regions that recognize SARS-CoV-2/2019-nCoV RNA genome. The heavy loading of rRT-PCR (qPCR) machine and handling labor have tight-packed the instruments as well as the manpower almost in every country. Therefore, the alternative approaches are eagerly waiting to be developed. In this review article, we sort out some state-of-the-art novel approaches that might be applied for a fast, sensitive, and precise detection of SARS-CoV-2/2019-nCoV not only to help the routine laboratory testing but also to improve effective quarantine.


Asunto(s)
Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/diagnóstico , Neumonía Viral/diagnóstico , Técnicas de Laboratorio Clínico , Humanos , Pandemias , ARN Viral/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos
4.
Monaldi Arch Chest Dis ; 90(2)2020 May 14.
Artículo en Inglés | MEDLINE | ID: mdl-32498503

RESUMEN

The enduring epidemic outbreak which started in Wuhan city of China, in December 2019 caused by the 2019 novel coronavirus (COVID- 19) or the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has created a dangerous and deadly Public Health disaster of International apprehension, with cases confirmed in several countries. This novel community health trouble is frightening the universe with clinical, psychological, emotional, collapse of health system and economical slowdown in each and every part of the world infecting nearly 200 countries. A highly virulent and pathogenic COVID-19 viral infection with incubation period ranging from two to fourteen days, transmitted by breathing of infected droplets or contact with infected droplets, belongs to the genus Coronavirus with its high mutation rate in the Coronaviridae. The likely probable primary reservoir could be bats, because genomic analysis discovered that SARSCoV-2 is phylogenetically interrelated to SARS-like bat viruses. The transitional resource of origin and transfer to humans is not known, however, the rapidly developing pandemic has confirmed human to human transfer. Approximately 1,016,128 reported cases, 211,615 recovered cases and 53,069 deaths of COVID-2019 have been reported to date (April 2, 2020). The symptoms vary from asymptomatic, low grade pyrexia, dry cough, sore throat, breathlessness, tiredness, body aches, fatigue, myalgia, nausea, vomiting, diarrhea, to severe consolidation and pneumonia, acute respiratory distress syndrome (ARDS) and multiple organ dysfunction leading to death with case fatality rate ranging from 2 to 3%.


Asunto(s)
Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/epidemiología , Brotes de Enfermedades , Neumonía Viral/epidemiología , Animales , Antivirales/administración & dosificación , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/terapia , Desarrollo de Medicamentos , Salud Global , Humanos , Pandemias , Neumonía Viral/diagnóstico , Neumonía Viral/terapia , Salud Pública , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos
5.
Respir Res ; 21(1): 146, 2020 Jun 11.
Artículo en Inglés | MEDLINE | ID: mdl-32527255

RESUMEN

BACKGROUND: Older age and elevated d-dimer are reported risk factors for coronavirus disease 2019 (COVID-19). However, whether early radiographic change is a predictor of fatality remains unknown. METHODS: We retrospectively reviewed records of all laboratory-confirmed patients admitted to a quarantine unit at Tongji Hospital, a large regional hospital in Wuhan, China, between January 31 and March 5, 2020. Confirmed cases were defined by positive RT-PCR detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in throat-swab specimens. Chest CT images were reviewed independently by two radiologists. The Tongji Hospital ethics committee approved this study. RESULTS: A total of 102 patients were confirmed to have SARS-CoV-2 infection. As of March 25, 85 confirmed patients were discharged, 15 died, and 2 remained hospitalized. When compared with survivors, non-survivors were older (median age, 69 [interquartile range, 58-77] vs. 55 [44-66], p = 0.003), and more likely to have decreased lymphocyte count (0.5 vs. 0.9 ×  109/L, p = 0.006), elevated lactate dehydrogenase (LDH) (569.0 vs. 272.0 U/L, p < 0.001), elevated d-dimer (> 1 µg/mL, 86% vs. 37%, p = 0.002) on admission. Older age and elevated LDH were independent risk factors for fatality in a multivariate regression model included the above variables. In a subset of patients with CT images within the first week, higher total severity score, and more involved lung lobes (5 involved lobes) in CT images within the first week were significantly associated with fatality. Moreover, in this subset of patients, higher total severity score was the only independent risk factor in a multivariate analysis incorporating the above mentioned variables. CONCLUSIONS: Older age, elevated LDH on admission, and higher severity score of CT images within the first week are potential predictors of fatality in adults with COVID-19. These predictors may help clinicians identify patients with a poor prognosis at an early stage.


Asunto(s)
Técnicas de Laboratorio Clínico/métodos , Infecciones por Coronavirus/diagnóstico por imagen , Infecciones por Coronavirus/mortalidad , Mortalidad Hospitalaria/tendencias , Pandemias/estadística & datos numéricos , Neumonía Viral/diagnóstico por imagen , Neumonía Viral/mortalidad , Radiografía Torácica/métodos , Adulto , Anciano , Anciano de 80 o más Años , Análisis de Varianza , China , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/terapia , Bases de Datos Factuales , Progresión de la Enfermedad , Femenino , Hospitalización/estadística & datos numéricos , Hospitales Públicos , Humanos , Unidades de Cuidados Intensivos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Pandemias/prevención & control , Neumonía Viral/prevención & control , Neumonía Viral/terapia , Valor Predictivo de las Pruebas , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , Análisis de Supervivencia
6.
PLoS One ; 15(6): e0234682, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32530929

RESUMEN

Novel Corona virus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or 2019-nCoV), and the subsequent disease caused by the virus (coronavirus disease 2019 or COVID-19), is an emerging global health concern that requires a rapid diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the standard for SARS-CoV-2 detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper field based testing at point-of-risk. The objective of this study was to develop a rapid screening diagnostic test that could be completed in 30-45 minutes. Simulated patient samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the SARS-CoV-2 nucleic sequence. RNA isolated from nasopharyngeal swabs collected from actual COVID-19 patients was also tested. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses. RT-LAMP specifically detected SARS-CoV-2 in both simulated patient samples and clinical specimens. This test was performed in 30-45 minutes. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts in the field and potential ports of entry.


Asunto(s)
Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , Neumonía Viral/diagnóstico , Pruebas en el Punto de Atención , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Betacoronavirus/genética , Infecciones por Coronavirus/virología , Cartilla de ADN , Humanos , Técnicas de Diagnóstico Molecular/economía , Técnicas de Diagnóstico Molecular/instrumentación , Técnicas de Amplificación de Ácido Nucleico/economía , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Pandemias , Neumonía Viral/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/economía , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Sensibilidad y Especificidad , Factores de Tiempo
7.
Gac Med Mex ; 156(3): 208-216, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32539016

RESUMEN

Introduction: As of March 23, 2020, suspension of non-essential activities was declared in Mexico throughout the country in order to mitigate the spread of the COVID-19 pandemic. Objective: To analyze data on the first 1,510 laboratory-confirmed cases of COVID-19 in Mexico, and to describe the geographical distribution of the disease and its transmission dynamics. Method: Description of the first COVID-19 cases with real-time RT-PCR-positive test, as well as evaluation of epidemiological measures, cumulative incidence, rate of transmission, and mortality and lethality rates during the first month of the epidemic. Results: Average age was 43 years, and 58 % were males; 44 % of initial cases were imported. Lethality in the population during the first month went from 1.08 to 3.97 per 100 cases; however, the trend is linear and similar to that observed in Europe. Conclusions: In Mexico, social distancing is being applied, but studies are still required on the dynamics of the epidemic, person-to-person transmission, incidence of subclinical infections, and patient survival.


Asunto(s)
Infecciones por Coronavirus/epidemiología , Neumonía Viral/epidemiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Aislamiento Social , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/transmisión , Femenino , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , México/epidemiología , Persona de Mediana Edad , Pandemias/prevención & control , Neumonía Viral/prevención & control , Neumonía Viral/transmisión , Sobrevida , Adulto Joven
8.
Emerg Microbes Infect ; 9(1): 1175-1179, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32448084

RESUMEN

Different primers/probes sets have been developed all over the world for the nucleic acid detection of SARS-CoV-2 by quantitative real time polymerase chain reaction (qRT-PCR) as a standard method. In our recent study, we explored the feasibility of droplet digital PCR (ddPCR) for clinical SARS-CoV-2 nucleic acid detection compared with qRT-PCR using the same primer/probe sets issued by Chinese Center for Disease Control and Prevention (CDC) targeting viral ORF1ab or N gene, which showed that ddPCR could largely minimize the false negatives reports resulted by qRT-PCR [Suo T, Liu X, Feng J, et al. ddPCR: a more sensitive and accurate tool for SARS-CoV-2 detection in low viral load specimens. medRxiv [Internet]. 2020;2020.02.29.20029439. Available from: https://medrxiv.org/content/early/2020/03/06/2020.02.29.20029439.abstract]. Here, we further stringently compared the performance of qRT-PCR and ddPCR for 8 primer/probe sets with the same clinical samples and conditions. Results showed that none of 8 primer/probe sets used in qRT-PCR could significantly distinguish true negatives and positives with low viral load (10-4 dilution). Moreover, false positive reports of qRT-PCR with UCDC-N1, N2 and CCDC-N primers/probes sets were observed. In contrast, ddPCR showed significantly better performance in general for low viral load samples compared to qRT-PCR. Remarkably, the background readouts of ddPCR are relatively lower, which could efficiently reduce the production of false positive reports.


Asunto(s)
Betacoronavirus/genética , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/virología , Reacción en Cadena de la Polimerasa Multiplex , Neumonía Viral/diagnóstico , Neumonía Viral/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Cartilla de ADN , Sondas de ADN , Humanos , Reacción en Cadena de la Polimerasa Multiplex/métodos , Pandemias , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Sensibilidad y Especificidad , Carga Viral
9.
Emerg Microbes Infect ; 9(1): 1259-1268, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32438868

RESUMEN

Quantitative real time PCR (RT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. However, due to the low viral load specimens and the limitations of RT-PCR, significant numbers of false negative reports are inevitable, which results in failure to timely diagnose, cut off transmission, and assess discharge criteria. To improve this situation, an optimized droplet digital PCR (ddPCR) was used for detection of SARS-CoV-2, which showed that the limit of detection of ddPCR is significantly lower than that of RT-PCR. We further explored the feasibility of ddPCR to detect SARS-CoV-2 RNA from 77 patients, and compared with RT-PCR in terms of the diagnostic accuracy based on the results of follow-up survey. 26 patients of COVID-19 with negative RT-PCR reports were reported as positive by ddPCR. The sensitivity, specificity, PPV, NPV, negative likelihood ratio (NLR) and accuracy were improved from 40% (95% CI: 27-55%), 100% (95% CI: 54-100%), 100%, 16% (95% CI: 13-19%), 0.6 (95% CI: 0.48-0.75) and 47% (95% CI: 33-60%) for RT-PCR to 94% (95% CI: 83-99%), 100% (95% CI: 48-100%), 100%, 63% (95% CI: 36-83%), 0.06 (95% CI: 0.02-0.18), and 95% (95% CI: 84-99%) for ddPCR, respectively. Moreover, 6/14 (42.9%) convalescents were detected as positive by ddPCR at 5-12 days post discharge. Overall, ddPCR shows superiority for clinical diagnosis of SARS-CoV-2 to reduce the false negative reports, which could be a powerful complement to the RT-PCR.


Asunto(s)
Betacoronavirus/genética , Infecciones por Coronavirus/diagnóstico , Neumonía Viral/diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacciones Falso Negativas , Humanos , Límite de Detección , Pandemias , ARN Viral/genética , Carga Viral/métodos
10.
J Med Microbiol ; 69(5): 705-711, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32369002

RESUMEN

Introduction. Pneumocystis jirovecii pneumonia (PCP) is a severe disease affecting immunocompromised patients. Diagnosis is difficult due to the low sensitivity of direct examination and inability to grow the pathogen in culture. Quantitative PCR in bronchoalveolar lavage fluid (BAL) has high sensitivity, but limited specificity for distinguishing PCP from colonization.Aim. To assess the performance of an in-house quantitative PCR to discriminate between PCP and colonization.Methodology. This was a single-centre retrospective study including all patients with a positive PCR result for P. jirovecii in BAL between 2009 and 2017. Irrespective of PCR results, PCP was defined as the presence of host factors and clinical/radiological criteria consistent with PCP and (i) the presence of asci at direct examination of respiratory sample or (ii) anti-PCP treatment initiated with clinical response and absence of alternative diagnosis. Colonization was considered for cases who did not receive anti-PCP therapy with a favourable outcome or an alternative diagnosis. Cases who did not meet the above mentioned criteria were classified as 'undetermined'.Results. Seventy-one patients with positive P. jirovecii PCR were included (90 % non-HIV patients). Cases were classified as follows: 37 PCP, 22 colonization and 12 undetermined. Quantitative PCR values in BAL were significantly higher in patients with PCP versus colonization or undetermined (P<0.0001). The cut-off of 5×103 copies/ml was able to discriminate PCP cases from colonization with 97 % sensitivity, 82 % specificity, 90 % positive predictive value and 95 % negative predictive value.Conclusions. Our quantitative PCR for P. jirovecii in BAL was reliable to distinguish PCP cases from colonization in this predominantly non-HIV population.


Asunto(s)
Pneumocystis carinii/clasificación , Pneumocystis carinii/genética , Neumonía por Pneumocystis/diagnóstico , Neumonía por Pneumocystis/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Adolescente , Adulto , Anciano , Algoritmos , Niño , Preescolar , Coinfección , Femenino , Humanos , Masculino , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular , Neumonía por Pneumocystis/tratamiento farmacológico , Neumonía por Pneumocystis/mortalidad , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Estudios Retrospectivos , Adulto Joven
11.
Infect Dis (Lond) ; 52(8): 571-574, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32420777

RESUMEN

Introduction: With the emergence of novel coronavirus disease 2019 (COVID-19) in many countries, medical resources currently focus on the treatment of confirmed patients and screening of suspected cases. Asymptomatic patients may be contagious, which makes epidemic control difficult. We describe an asymptomatic patient with a positive real-time polymerase chain reaction (RT-PCR) test in urine.Case report: An asymptomatic girl was identified during the epidemiological investigation of a confirmed COVID-19 patient. When admitted to the hospital on 24 February 2020, she had no clinical manifestations. A throat swab was negative for RT-PCR, but urine was positive. She was given antiviral and symptomatic supportive treatment. On 26 February, a throat swab RT-PCR was positive. RT-PCR in throat swabs and urine were negative on 3 and 5 March, and on 9 and 12 March, throat swabs were still negative. At follow-up on 26 March, she felt well, throat swab RT-PCR was negative, and isolation was lifted.Conclusion: The urine of asymptomatic patients may be contagious. RT-PCR in urine might be a useful supplement in screening when the RT-PCR is negative in throat swabs.


Asunto(s)
Infecciones Asintomáticas , Betacoronavirus/aislamiento & purificación , Técnicas de Laboratorio Clínico/métodos , Infecciones por Coronavirus/orina , Neumonía Viral/orina , Adolescente , Betacoronavirus/genética , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/virología , Femenino , Humanos , Pandemias , Neumonía Viral/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Orina/virología
12.
RNA ; 26(7): 771-783, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32358057

RESUMEN

The current COVID-19 pandemic presents a serious public health crisis, and a better understanding of the scope and spread of the virus would be aided by more widespread testing. Nucleic-acid-based tests currently offer the most sensitive and early detection of COVID-19. However, the "gold standard" test pioneered by the U.S. Centers for Disease Control and Prevention takes several hours to complete and requires extensive human labor, materials such as RNA extraction kits that could become in short supply, and relatively scarce qPCR machines. It is clear that a huge effort needs to be made to scale up current COVID-19 testing by orders of magnitude. There is thus a pressing need to evaluate alternative protocols, reagents, and approaches to allow nucleic-acid testing to continue in the face of these potential shortages. There has been a tremendous explosion in the number of papers written within the first weeks of the pandemic evaluating potential advances, comparable reagents, and alternatives to the "gold-standard" CDC RT-PCR test. Here we present a collection of these recent advances in COVID-19 nucleic acid testing, including both peer-reviewed and preprint articles. Due to the rapid developments during this crisis, we have included as many publications as possible, but many of the cited sources have not yet been peer-reviewed, so we urge researchers to further validate results in their own laboratories. We hope that this review can urgently consolidate and disseminate information to aid researchers in designing and implementing optimized COVID-19 testing protocols to increase the availability, accuracy, and speed of widespread COVID-19 testing.


Asunto(s)
Betacoronavirus/aislamiento & purificación , Técnicas de Laboratorio Clínico , Técnicas de Amplificación de Ácido Nucleico/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Betacoronavirus/genética , Sistemas CRISPR-Cas , Centers for Disease Control and Prevention, U.S. , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Infecciones por Coronavirus/diagnóstico , Humanos , Nasofaringe/virología , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Sistemas de Atención de Punto , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Factores de Tiempo , Estados Unidos , Flujo de Trabajo
13.
J Card Surg ; 35(6): 1342-1344, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32400044

RESUMEN

While elective cardiac surgeries have been postponed to prevent coronavirus disease 2019 (COVID-19) transmission and to reduce resource utilization, patients with urgent indications necessitating surgery may still be at risk of contracting the disease throughout their postoperative recovery. We present a case of an 81-year-old female who underwent urgent coronary artery bypass grafting and was readmitted following discharge to a nursing facility with a cluster of COVID-19 cases. Despite symptomatology and imaging concerning for COVID-19, two initial reverse transcription polymerase chain reaction (RT-PCR) tests were negative but a third test was positive. This case emphasizes the risks of discharge location in the COVID-19 era as well as the importance of clinical suspicion, early isolation practices for those presumed positive, and repeat testing, given the marginal sensitivity of available COVID-19 RT-PCR.


Asunto(s)
Técnicas de Laboratorio Clínico/métodos , Técnicas de Laboratorio Clínico/estadística & datos numéricos , Puente de Arteria Coronaria/métodos , Infecciones por Coronavirus/diagnóstico , Transmisión de Enfermedad Infecciosa/prevención & control , Aislamiento de Pacientes , Neumonía Viral/diagnóstico , Anciano de 80 o más Años , Betacoronavirus , Angiografía Coronaria/métodos , Reacciones Falso Negativas , Femenino , Estudios de Seguimiento , Hogares para Ancianos , Humanos , Evaluación de Necesidades , Casas de Salud , Pandemias , Alta del Paciente , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reproducibilidad de los Resultados , Medición de Riesgo
14.
Langenbecks Arch Surg ; 405(3): 353-355, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32385569

RESUMEN

PURPOSE: COVID-19 greatly affected millions and affected the way we practice with heightened posture in the way we treat surgical patients. Surgical consensus guidelines are recommending caution in the use of laparoscopy for the theoretical possibility of viral transmission from aerosolization of tissue and peritoneal fluid during surgery. However, there has yet to be proof of COVID-19 being present in peritoneal fluid, justifying the consensus statements. We aim to assess the presence of COVID-19 in peritoneal fluid. METHODS: We performed a laparoscopic appendicectomy for a COVID-19-infected patient with acute appendicitis. Peritoneal fluid and peritoneal washings were collected and sent for COVID-19 PCR. RESULTS: The peritoneal fluid sample collected on entry and at the end of the operation was negative for COVID-19 on PCR. The patient had an uneventful recovery from surgery. CONCLUSIONS: This case revealed that COVID-19 was not detected in peritoneal fluid and peritoneal washings in a patient infected with COVID-19. This study provides novel preliminary data in the investigation of COVID-19 transmission from laparoscopy-related aerosolization.


Asunto(s)
Apendicectomía/métodos , Apendicitis/cirugía , Líquido Ascítico/virología , Infecciones por Coronavirus/diagnóstico , Transmisión de Enfermedad Infecciosa de Paciente a Profesional/prevención & control , Neumonía Viral/diagnóstico , Apendicitis/diagnóstico , Técnicas de Laboratorio Clínico/métodos , ADN Viral/aislamiento & purificación , Reacciones Falso Negativas , Estudios de Seguimiento , Humanos , Laparoscopía/efectos adversos , Laparoscopía/métodos , Masculino , Salud Laboral , Pandemias , Seguridad del Paciente , Lavado Peritoneal/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Medición de Riesgo , Resultado del Tratamiento , Adulto Joven
15.
BMC Infect Dis ; 20(1): 362, 2020 May 24.
Artículo en Inglés | MEDLINE | ID: mdl-32448123

RESUMEN

BACKGROUND: The Xpert MTB/RIF assay is an automated molecular test that is designed to simultaneously detect Mycobacterium tuberculosis (MTB) complex and rifampin resistance. However, there are relatively few studies on this method in China. Xpert has been routinely used at Peking University People's Hospital (PKUPH) since November 2016. Thus, the aim of this study was to evaluate the performance of Xpert, and provide a reference and guidance for the detection and diagnosis of TB in non-TB specialized hospitals. METHODS: The medical records of inpatients simultaneously tested with Xpert, acid-fast bacilli (AFB) smear microscopy, and interferon-gamma release assay (IGRA, by T-SPOT®.TB) at PKUPH from November 2016 to October 2018 were reviewed. Active TB cases were considered according to a composite reference standard (CRS). Then, the three methods were evaluated and compared. RESULTS: In total, 787 patients simultaneously tested with Xpert, AFB, and IGRA were enrolled; among them 11.3% (89/787) were diagnosed and confirmed active pulmonary TB (PTB, 52 cases), extrapulmonary TB (EPTB, 17 cases), and tuberculous pleurisy (TP, 20 cases). The sensitivity of Xpert in detecting PTB, EPTB, and TP was 88.5, 76.5, and 15.0%, respectively, which was slightly lower than IGRA (96.2, 82.4, and 95.0%, respectively), but higher than AFB (36.5, 11.8, and 0%, respectively); IGRA showed the highest sensitivity, but its specificity (55.9, 67.1, and 45.2%, respectively) was significantly lower than Xpert (99.6, 99.4, and 100%, respectively) and AFB (99.0, 99.4, and 100%, respectively) (P < 0.001). The sensitivity of Xpert in detecting lung tissue, cerebrospinal fluid, lymph nodes, and joint fluid was 100%, followed by sputum (88.5%), alveolar lavage (85.7%), and bronchoscopy secretion (81.2%); the pleural fluid sensitivity was the lowest, only 15.0%. For AFB negative patients, the sensitivity of Xpert in detecting PTB, EPTB, and TP was 84.9, 73.3, and 15.0%, respectively. CONCLUSIONS: Xpert showed both high sensitivity and high specificity, and suggested its high value in TB diagnosis; however, the application of pleural fluid is still limited, and should be improved. Owing to the high sensitivity of IGRA, it is recommended for use as a supplementary test, especially for assisting in the diagnosis of TP and EPTB.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Rifampin/farmacología , Tuberculosis Pleural/diagnóstico , Tuberculosis Pulmonar/diagnóstico , Adolescente , Adulto , Anciano , China , Femenino , Hospitales de Enseñanza , Humanos , Ensayos de Liberación de Interferón gamma/métodos , Masculino , Microscopía/métodos , Persona de Mediana Edad , Estudios Retrospectivos , Sensibilidad y Especificidad , Tuberculosis Pleural/microbiología , Tuberculosis Pulmonar/microbiología , Adulto Joven
16.
Clin Chim Acta ; 507: 139-142, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32335089

RESUMEN

BACKGROUND: The detection of SARS-CoV-2 RNA by real-time reverse transcription-polymerase chain reaction (rRT-PCR) is used to confirm the clinical diagnosis of COVID-19 by molecular diagnostic laboratories. We developed a multiplex rRT-PCR methodology for the detection of SARS-CoV-2 RNA. METHODS: Three genes were used for multiplex rRT-PCR: the Sarbecovirus specific E gene, the SARS-CoV-2 specific N gene, and the human ABL1 gene as an internal control. RESULTS: Good correlation of Cq values was observed between the simplex and multiplex rRT-PCR methodologies. Low copies (<25 copies/reaction) of SARS-CoV-2 RNA were detected by the novel multiplex rRT-PCR method. CONCLUSION: The proposed multiplex rRT-PCR methodology will enable highly sensitive detection of SARS-CoV-2 RNA, reducing reagent use and cost, and time required by clinical laboratory technicians.


Asunto(s)
Betacoronavirus/genética , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/genética , Neumonía Viral/diagnóstico , Neumonía Viral/genética , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Betacoronavirus/aislamiento & purificación , Técnicas de Laboratorio Clínico/métodos , Técnicas de Laboratorio Clínico/normas , Humanos , Pandemias , ARN Viral/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reproducibilidad de los Resultados , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Esputo/química , Esputo/virología
17.
Clin Chim Acta ; 507: 164-166, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32343948

RESUMEN

Validation studies of serological antibody tests must be properly designed for clinical, epidemiological and Public Health objectives such as confirmation of suspected COVID-19 cases, certification of seroconversion after infection, and epidemiological surveillance. We evaluated the kinetics of IgM, IgA and IgG SARS-CoV-2 antibodies in COVID-19 patients with confirmed (rRT-PCR) infection. We found that the IgA response appears and grows early, peaks at week 3, and it is stronger and more persistent than the IgM response. Further longitudinal investigations of virus-specific antibodies functions and of their protective efficacy over time are needed.


Asunto(s)
Anticuerpos Antivirales/sangre , Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/sangre , Infecciones por Coronavirus/diagnóstico , Glicoproteínas/sangre , Inmunoglobulina A/sangre , Neumonía Viral/sangre , Neumonía Viral/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Estudios Longitudinales , Mediciones Luminiscentes/métodos , Masculino , Persona de Mediana Edad , Pandemias , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adulto Joven
18.
Clin Chem Lab Med ; 58(7): 1089-1094, 2020 06 25.
Artículo en Inglés | MEDLINE | ID: mdl-32301745

RESUMEN

Objectives In December 2019, a novel coronavirus (SARS-CoV-2)-infected pneumonia (COVID-19) occurred in Wuhan, China. Laboratory-based diagnostic tests utilized real-time reverse transcriptase polymerase chain reaction (RT-PCR) on throat samples. This study evaluated the diagnostic value to analyzing throat and sputum samples in order to improve accuracy and detection efficiency. Methods Paired specimens of throat swabs and sputum were obtained from 54 cases, and RNA was extracted and tested for 2019-nCoV (equated with SARS-CoV-2) by the RT-PCR assay. Results The positive rates of 2019-nCoV from sputum specimens and throat swabs were 76.9% and 44.2%, respectively. Sputum specimens showed a significantly higher positive rate than throat swabs in detecting viral nucleic acid using the RT-PCR assay (p = 0.001). Conclusions The detection rates of 2019-nCoV from sputum specimens were significantly higher than those from throat swabs. We suggest that sputum would benefit for the detection of 2019-nCoV in patients who produce sputum. The results can facilitate the selection of specimens and increase the accuracy of diagnosis.


Asunto(s)
Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/genética , Neumonía Viral/diagnóstico , Neumonía Viral/genética , Manejo de Especímenes/métodos , Adulto , Anciano , Anciano de 80 o más Años , Betacoronavirus/genética , Betacoronavirus/patogenicidad , China , Técnicas de Laboratorio Clínico/métodos , Coronavirus/genética , Coronavirus/patogenicidad , Pruebas Diagnósticas de Rutina/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pandemias , Faringe/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Virus del SRAS/genética , Virus del SRAS/patogenicidad , Esputo
19.
Clin Chem Lab Med ; 58(7): 1095-1099, 2020 06 25.
Artículo en Inglés | MEDLINE | ID: mdl-32301746

RESUMEN

Objectives The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to date, the epidemic has gradually spread to 209 countries worldwide with more than 1.5 million infected people and 100,000 deaths. Amplification of viral RNA by rRT-PCR serves as the gold standard for confirmation of infection, yet it needs a long turnaround time (3-4 h to generate results) and shows false-negative rates as large as 15%-20%. In addition, the need of certified laboratories, expensive equipment and trained personnel led many countries to limit the rRT-PCR tests only to individuals with pronounced respiratory syndrome symptoms. Thus, there is a need for alternative, less expensive and more accessible tests. Methods We analyzed the plasma levels of white blood cells (WBCs), platelets, C-reactive protein (CRP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), alkaline phosphatase and lactate dehydrogenase (LDH) of 207 patients who, after being admitted to the emergency room of the San Raffaele Hospital (Milan, Italy) with COVID-19 symptoms, were rRT-PCR tested. Of them, 105 tested positive, whereas 102 tested negative. Results Statistically significant differences were observed for WBC, CRP, AST, ALT and LDH. Empirical thresholds for AST and LDH allowed the identification of 70% of either COVID-19-positive or -negative patients on the basis of routine blood test results. Conclusions Combining appropriate cutoffs for certain hematological parameters could help in identifying false-positive/negative rRT-PCR tests. Blood test analysis might be used as an alternative to rRT-PCR for identifying COVID-19-positive patients in those countries which suffer from a large shortage of rRT-PCR reagents and/or specialized laboratory.


Asunto(s)
Biomarcadores/sangre , Infecciones por Coronavirus/diagnóstico , Pruebas Hematológicas/métodos , Neumonía Viral/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Alanina Transaminasa/análisis , Alanina Transaminasa/sangre , Fosfatasa Alcalina/análisis , Fosfatasa Alcalina/sangre , Aspartato Aminotransferasas/análisis , Aspartato Aminotransferasas/sangre , Betacoronavirus/patogenicidad , Plaquetas , Proteína C-Reactiva/análisis , Infecciones por Coronavirus/sangre , Femenino , Humanos , Italia , L-Lactato Deshidrogenasa/análisis , L-Lactato Deshidrogenasa/sangre , Laboratorios , Leucocitos , Masculino , Persona de Mediana Edad , Pandemias , Neumonía Viral/sangre , ARN Viral , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Estudios Retrospectivos , gamma-Glutamiltransferasa/análisis , gamma-Glutamiltransferasa/sangre
20.
PLoS Negl Trop Dis ; 14(4): e0008087, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-32330127

RESUMEN

There is growing interest in local elimination of soil-transmitted helminth (STH) infection in endemic settings. In such settings, highly sensitive diagnostics are needed to detect STH infection. We compared double-slide Kato-Katz, the most commonly used copromicroscopic detection method, to multi-parallel quantitative polymerase chain reaction (qPCR) in 2,799 stool samples from children aged 2-12 years in a setting in rural Bangladesh with predominantly low STH infection intensity. We estimated the sensitivity and specificity of each diagnostic using Bayesian latent class analysis. Compared to double-slide Kato-Katz, STH prevalence using qPCR was almost 3-fold higher for hookworm species and nearly 2-fold higher for Trichuris trichiura. Ascaris lumbricoides prevalence was lower using qPCR, and 26% of samples classified as A. lumbricoides positive by Kato-Katz were negative by qPCR. Amplicon sequencing of the 18S rDNA from 10 samples confirmed that A. lumbricoides was absent in samples classified as positive by Kato-Katz and negative by qPCR. The sensitivity of Kato-Katz was 49% for A. lumbricoides, 32% for hookworm, and 52% for T. trichiura; the sensitivity of qPCR was 79% for A. lumbricoides, 93% for hookworm, and 90% for T. trichiura. Specificity was ≥ 97% for both tests for all STH except for Kato-Katz for A. lumbricoides (specificity = 68%). There were moderate negative, monotonic correlations between qPCR cycle quantification values and eggs per gram quantified by Kato-Katz. While it is widely assumed that double-slide Kato-Katz has few false positives, our results indicate otherwise and highlight inherent limitations of the Kato-Katz technique. qPCR had higher sensitivity than Kato-Katz in this low intensity infection setting.


Asunto(s)
Helmintiasis/diagnóstico , Parasitosis Intestinales/diagnóstico , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Ancylostomatoidea/aislamiento & purificación , Animales , Ascaris lumbricoides/aislamiento & purificación , Bangladesh , Niño , Preescolar , ADN de Helmintos/genética , ADN Ribosómico/genética , Heces/parasitología , Femenino , Humanos , Lactante , Masculino , ARN Ribosómico 18S/genética , Población Rural , Sensibilidad y Especificidad , Trichuris/aislamiento & purificación
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