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1.
J Transl Med ; 19(1): 30, 2021 01 07.
Artículo en Inglés | MEDLINE | ID: mdl-33413461

RESUMEN

BACKGROUND: COVID-19 has caused a global pandemic and the death toll is increasing. However, there is no definitive information regarding the type of clinical specimens that is the best for SARS-CoV-2 detection, the antibody levels in patients with different duration of disease, and the relationship between antibody level and viral load. METHODS: Nasopharyngeal swabs, anal swabs, saliva, blood, and urine specimens were collected from patients with a course of disease ranging from 7 to 69 days. Viral load in different specimen types was measured using droplet digital PCR (ddPCR). Meanwhile, anti-nucleocapsid protein (anti-N) IgM and IgG antibodies and anti-spike protein receptor-binding domain (anti-S-RBD) IgG antibody in all serum samples were tested using ELISA. RESULTS: The positive detection rate in nasopharyngeal swab was the highest (54.05%), followed by anal swab (24.32%), and the positive detection rate in saliva, blood, and urine was 16.22%, 10.81%, and 5.41%, respectively. However, some patients with negative nasopharyngeal swabs had other specimens tested positive. There was no significant correlation between antibody level and days after symptoms onset or viral load. CONCLUSIONS: Other specimens could be positive in patients with negative nasopharyngeal swabs, suggesting that for patients in the recovery period, specimens other than nasopharyngeal swabs should also be tested to avoid false negative results, and anal swabs are recommended. The antibody level had no correlation with days after symptoms onset or the viral load of nasopharyngeal swabs, suggesting that the antibody level may also be affected by other factors.


Asunto(s)
Anticuerpos Antivirales/sangre , /virología , /aislamiento & purificación , Carga Viral , Adulto , Anciano , Anciano de 80 o más Años , Canal Anal/virología , Sangre/virología , China/epidemiología , Reacciones Falso Negativas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nasofaringe/virología , Pandemias , Saliva/virología , Manejo de Especímenes , Factores de Tiempo , Investigación en Medicina Traslacional , Orina/virología
3.
J Clin Pathol ; 74(1): 64-66, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32482888

RESUMEN

COVID-19 is a zoonotic viral infection that originated in Wuhan, China, in late 2019. WHO classified the resulting pandemic as a 'global health emergency' due to its virulence and propensity to cause acute respiratory distress syndrome. The COVID-19 pandemic has had a major impact on diagnostic laboratories, particularly those handling cell and tissue specimens. This development carries serious implications for laboratory practice in that safety of personnel has to be balanced against high-quality analysis and timely reporting of results. The aim of this article is to present some recommendations for the handling of such specimens in the preanalytical, analytical and postanalytical phases of laboratory testing and analysis in an era of high COVID-19 prevalence, such as that seen, for example, in the UK, Spain, Italy and France.


Asunto(s)
Infección de Laboratorio/prevención & control , Salud Laboral , Patología Clínica/métodos , Manejo de Especímenes/métodos , Europa (Continente) , Humanos , Laboratorios
4.
Diagn Microbiol Infect Dis ; 99(1): 115206, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33045499

RESUMEN

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.


Asunto(s)
/métodos , /aislamiento & purificación , Manejo de Especímenes/métodos , /normas , Genes Virales/genética , Humanos , India/epidemiología , Nasofaringe/virología , Faringe/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Sensibilidad y Especificidad , Manejo de Especímenes/normas , Centros de Atención Terciaria
5.
J Infect Chemother ; 27(1): 126-129, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33060046

RESUMEN

Considering the issues of shortage of medical resources and the invasiveness and infection risk involved in the collection of nasopharyngeal swab specimens, there is a need for an effective alternative test specimen for SARS-CoV-2 RNA detection. Here, we investigated suitability of saliva as a non-invasively obtained specimen for molecular detection of SARS-CoV-2 RNA in Japanese patients with COVID-19. In total, 28 paired clinical specimens of saliva and nasopharyngeal swabs were collected from 12 patients at various time points after symptom onset. Each specimen was assayed using reverse transcription real-time polymerase chain reaction (rRT-PCR) on the BD MAX open system using primers and probes targeting the N-gene. The saliva and nasopharyngeal swab specimens showed 19 and 15 positive results, respectively. No invalid (PCR inhibition) result was observed for any specimen. The qualitative results of each specimen obtained in the period immediately after symptom onset were similar. Three convalescent patients presented saliva-positive results, whereas their nasopharyngeal swabs were negative at four different time points, suggesting that saliva may be superior to nasopharyngeal swabs in terms of obtaining stable assay result of SARS-CoV-2. In conclusion, our results suggest that saliva can potentially serve as an alternative to nasopharyngeal swabs as a specimen for SARS-CoV-2 rRT-PCR. As saliva can be collected by patients themselves, it may be an effective way to overcome the shortage of personal protective equipment and specimen sampling tools.


Asunto(s)
Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/diagnóstico , Nasofaringe/virología , Neumonía Viral/diagnóstico , ARN Viral/aislamiento & purificación , Saliva/virología , Técnicas de Laboratorio Clínico , Humanos , Japón , Pandemias , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Manejo de Especímenes/métodos
6.
Diagn Microbiol Infect Dis ; 99(1): 115209, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33080426

RESUMEN

Nasopharyngeal flocked swabs placed in viral transport media (VTM) are the preferred collection methodology for respiratory virus testing. Due to the rapid depletion of available reagents and swabs, we have validated an alternative swab placed in phosphate-buffered saline (PBS) for use in respiratory virus testing in a SARS-CoV-2 real-time polymerase chain reaction assay and a multiplexed respiratory virus panel. We collected nasopharyngeal (NP) swabs and oropharyngeal (OP) swabs from 10 healthy volunteers. Flocked swabs were placed in VTM and alternative swabs in PBS. In this feasibility study, we show that NP collection is better for detection of human material than OP collection, as measured by significantly lower RNase P gene cycle threshold values, and that a Dacron polyester swab in PBS shows equivalent detection of SARS-CoV-2 and RSV to a flocked swab in VTM in contrived specimens. Diluted SARS-CoV-2-positive patient specimens are detectable for up to 72 h at 4 °C.


Asunto(s)
/diagnóstico , Manejo de Especímenes/métodos , Medios de Cultivo , Estudios de Factibilidad , Humanos , Nasofaringe/virología , Orofaringe/virología , Tereftalatos Polietilenos , Virus Sincitial Respiratorio Humano/genética , Virus Sincitial Respiratorio Humano/aislamiento & purificación , /genética
7.
Ann Anat ; 233: 151608, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33022405

RESUMEN

COVID-19 is an infection caused by the SARS-CoV-2 virus, initially identified in the city of Wuhan, China, in December 2019. Since then, the virus has spread to the continents, causing a major pandemic. The impacts of this pandemic on the education of human anatomy interfere in at least two aspects: (1) receiving and managing anatomical specimens in anatomy laboratories and (2) adaptations for classes on remote virtual teaching. Therefore, this study reviewed and discussed the legal and bioethical aspects, considering the scenario of a South American Country, aiming to stimulate the debate on these two relevant themes in the international community. Because of the COVID-19 pandemic and the impossibility of mass testing, anatomists and other workers in the field must deal with the risk of receiving bodies infected with SARS-CoV-2. In this situation, additional care measures in biosafety practices are essential to protect the staff. Such measures are: the bodies must be preserved by the perfusion of formaldehyde or other fixative solutions; embalming must be performed in ventilated rooms with a good air exhaust system; to avoid excessive manipulation of bodies and procedures such as pulmonary insufflation or craniotomy; and proper use of personal protective equipment, including lab coat, gloves and masks. As for exposure of body images in online classes, this review showed that there are no legal impediments to this end. However, anatomists must adopt measures aimed at protecting the memory of the deceased, such as using secure digital platforms with restricted access; family authorization/consent and student awareness.


Asunto(s)
Anatomía/educación , Pandemias , Manejo de Especímenes/ética , Brasil , Contención de Riesgos Biológicos , Embalsamiento , Personal de Salud , Humanos , Legislación Médica , Seguridad , Ventilación
8.
J Virol Methods ; 287: 113969, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32918932

RESUMEN

The current COVID-19 pandemic constitutes a threat to the population worldwide with over 21 million infected people. There is an urgent need for the development of rapid and massive detection tools as well as the identification and isolation of infected individuals. we sought to evaluate different RT-qPCR kits and protocols to evaluate the best approach to be used omitting an RNA extraction step. We have investigated the sensitivity and performance of different commercially available RT-qPCR kits in detecting SARS-CoV-2 using 80 extracted RNA and NSS from COVID-19 diagnosed patients. We evaluated the ability of each kit to detect viral RNA from both kit-extracted or directly from a pre-boiled NSS observing that direct RNA detection is possible when Ct values are lower than 30 with the three kits tested. Since SARS-CoV-2 testing in most locations occurs once COVID-19 symptoms are evident and, therefore, viral loads are expected to be high, our protocol will be useful in supporting SARS-CoV-2 diagnosis, especially in America where COVID-19 cases have exploded in the recent weeks as well as in low- and middle-income countries, which would not have massive access to kit-based diagnosis. The information provided in this work paves the way for the development of more efficient SARS-CoV-2 detection approaches avoiding an RNA extraction step.


Asunto(s)
/métodos , /aislamiento & purificación , Manejo de Especímenes/métodos , /virología , Humanos , Nasofaringe/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Juego de Reactivos para Diagnóstico , Sensibilidad y Especificidad , Factores de Tiempo , Carga Viral
9.
Lab Med ; 52(1): e15-e16, 2021 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-33206195

RESUMEN

OBJECTIVE: During the current pandemic, COVID-19 has been detected in patients using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) that confirms the presence of SARS-CoV-2 RNA. The demand for increased testing, particularly for asymptomatic individuals required alternative approaches to single-patient RT-PCR testing, such as pooling. METHODS: This study explored the impact of dilution on the detectability of SARS-CoV-2 in asymptomatic patients using RT-PCR and demonstrated that pooling can be effective in low prevalence populations. RESULTS: The RT-PCR results for the 3:1, 5:1, and 7:1 aliquot samples showed little differences in CT values, confirming detection capability at these dilutions. CONCLUSION: Based on the results of the present study, a pooled approach with up to 5:1 sample aliquots and using the current RT-PCR methodology likely will detect SARS CoV2 RNA among asymptomatic patients.


Asunto(s)
/métodos , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , /genética , Enfermedades Asintomáticas , Tampones (Química) , Humanos , Manejo de Especímenes/métodos
10.
CMAJ ; 193(1): E1-E9, 2021 01 04.
Artículo en Inglés | MEDLINE | ID: mdl-33234533

RESUMEN

BACKGROUND: Research involving children with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has primarily focused on those presenting to emergency departments. We aimed to determine the symptoms most commonly associated with a positive result for a SARS-CoV-2 swab among community-based children. METHODS: We conducted an observational study among children tested and followed for SARS-CoV-2 infection using nasal, nasopharyngeal, throat or other (e.g., nasopharyngeal aspirate or tracheal secretions, or unknown) swabs between Apr. 13 and Sept. 30, 2020, in Alberta. We calculated positive likelihood ratios (LRs) for self-reported symptoms and a positive SARS-CoV-2 swab result in the entire cohort and in 3 sensitivity analyses: all children with at least 1 symptom, all children tested because of contact tracing whether they were symptomatic or not and all children 5 years of age or older. RESULTS: We analyzed results for 2463 children who underwent testing for SARS-CoV-2 infection; 1987 children had a positive result and 476 had a negative result. Of children with a positive test result for SARS-CoV-2, 714 (35.9%) reported being asymptomatic. Although cough (24.5%) and rhinorrhea (19.3%) were 2 of the most common symptoms among children with SARS-CoV-2 infection, they were also common among those with negative test results and were not predictive of a positive test (positive LR 0.96, 95% confidence interval [CI] 0.81-1.14, and 0.87, 95% CI 0.72-1.06, respectively). Anosmia/ageusia (positive LR 7.33, 95% CI 3.03-17.76), nausea/vomiting (positive LR 5.51, 95% CI 1.74-17.43), headache (positive LR 2.49, 95% CI 1.74- 3.57) and fever (positive LR 1.68, 95% CI 1.34-2.11) were the symptoms most predictive of a positive result for a SARS-CoV-2 swab. The positive LR for the combination of anosmia/ageusia, nausea/vomiting and headache was 65.92 (95% CI 49.48-91.92). INTERPRETATION: About two-thirds of the children who tested positive for SARS-CoV-2 infection reported symptoms. The symptoms most strongly associated with a positive SARS-CoV-2 swab result were anosmia/ageusia, nausea/vomiting, headache and fever.


Asunto(s)
/diagnóstico , Técnicas de Laboratorio Clínico/métodos , Pandemias , Manejo de Especímenes/métodos , Adolescente , Alberta/epidemiología , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Masculino
11.
Eur J Clin Microbiol Infect Dis ; 40(1): 193-195, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32666481

RESUMEN

PURPOSE: PCR on a nasopharyngeal sample is the reference method for the detection of SARS-nCoV-2. However, combined throat/nasal sampling as a testing method has several advantages. We compared the combined throat/nasal sampling with nasopharyngeal sampling for detection of SARS-CoV-2 in healthcare workers suspected of COVID-19. METHODS: In 107 healthcare workers with symptoms of COVID-19, combined throat/nasal sampling and nasopharyngeal sampling was performed. Detection of SARS-CoV-2 was performed by RT-PCR targeting. RESULTS: A total of 80 healthcare workers (74.8%) tested negative with both sampling methods, and 25 healthcare workers (23.4%) tested positive with both sampling methods. There were two discrepant results with positive PCR in combined throat/nasal swabs and negative PCR in nasopharyngeal swabs (1.9%). The κ index for concordance between the 2 sampling methods was high (0.95). The median cycle threshold (Ct) value of PCR on nasopharyngeal samples was significantly lower than the Ct value of PCR on combined throat/nasal samples (19 (IQR 17-20) versus 21 (IQR 18-29) cycles, p value 0.01). CONCLUSION: Combined throat/nasal swabs yield a similar sensitivity to detect SARS-CoV-2 as nasopharyngeal swabs and are a good alternative sampling method, despite a lower Ct value for the nasopharyngeal samples.


Asunto(s)
/métodos , Nasofaringe/virología , Nariz/virología , Faringe/virología , Manejo de Especímenes/métodos , Adulto , Personal de Salud/estadística & datos numéricos , Humanos , Persona de Mediana Edad , ARN Viral/aislamiento & purificación , Adulto Joven
12.
J Med Virol ; 93(2): 719-725, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-32706393

RESUMEN

Testing is one of the commendable measures for curbing the spread of coronavirus disease (COVID-19). But, it should be done using the most appropriate specimen and an accurate diagnostic test such as real-time reverse transcription-polymerase chain reaction (qRT-PCR). Therefore, a systematic review was conducted to determine the positive detection rate of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in different clinical specimens using qRT-PCR. A total of 8136 pooled clinical specimens were analyzed to detect SARS-CoV-2, the majority were nasopharyngeal swabs (69.6%). A lower respiratory tract (LRT) specimens had a positive rate (PR) of 71.3% (95% confidence interval [CI]: 60.3%-82.3%) while no virus was detected in the urinogenital specimens. Bronchoalveolar lavage fluid (BLF) specimen had the PR of 91.8% (95% CI: 79.9%-103.7%), followed by rectal swabs; 87.8% (95% CI: 78.6%-96.9%) then sputum; 68.1% (95% CI: 56.9%-79.4%). A low PR was observed in oropharyngeal swabs; 7.6% (95% CI: 5.7%-9.6%) and blood samples; 1.0% (95% CI: -0.1%-2.1%) whereas no SARS-CoV-2 was detected in urine samples. Feces had a PR of 32.8% (95% CI:1 5.8%-49.8%). Nasopharyngeal swab, a widely used specimen had a PR of 45.5% (95% CI: 31.2%-59.7%). In this study, SARS-CoV-2 was highly detected in LRT specimens while no virus was detected in urinogenital specimens. BLF had the highest PR followed by rectal swab then sputum. Nasopharyngeal swab which is widely used had moderate PR. Low PR was recorded in oropharyngeal swab and blood samples while no virus was found in urine samples. Last, the virus was detected in feces, suggesting SARS-CoV-2 transmission by the fecal route.


Asunto(s)
/métodos , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/estadística & datos numéricos , /aislamiento & purificación , Líquido del Lavado Bronquioalveolar/virología , /orina , Heces/virología , Humanos , Nasofaringe/virología , Orofaringe/virología , Manejo de Especímenes/métodos , Esputo/virología
13.
Diagn Microbiol Infect Dis ; 99(1): 115205, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33190092

RESUMEN

Pooling of 1 positive sample with up to 5 negative samples prior to testing with the Cepheid GenXpert SARS-CoV-2 assay did not adversely impact detection of positive samples. At our current prevalence of 2%, it could save up to 70% of the test kits.


Asunto(s)
/métodos , /aislamiento & purificación , Manejo de Especímenes/métodos , Humanos , Juego de Reactivos para Diagnóstico/economía , Juego de Reactivos para Diagnóstico/provisión & distribución , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
14.
Clin Chim Acta ; 512: 58-62, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33285119

RESUMEN

OBJECTIVE: It is unclear if implementation of biosafety action plans in response to the COVID-19 pandemic has affected laboratory quality metrics. METHODS: This retrospective study used quality data, including turnaround time (TAT) and number/type of unacceptable specimens from a stat laboratory supporting an outpatient medical clinic serving predominantly elderly cancer patients. Four months of data from the height of the COVID-19 pandemic (March-June 2020) were compared to the same months in 2019. RESULTS: March-May 2020 test volumes were decreased compared to 2019. June 2020 test volume was slightly increased compared to 2019. TATs in 2020 were similar/ slightly improved compared to the same months in 2019, due to shortened collect to receive and receive to verify TATs. The number and types of unacceptable specimens were similar in 2020 and 2019. CONCLUSIONS: Despite the challenges to the system caused by the pandemic, laboratory quality metrics were maintained.


Asunto(s)
Laboratorios/estadística & datos numéricos , Anciano , Contención de Riesgos Biológicos/normas , Humanos , Laboratorios/normas , Neoplasias , Pandemias , Estudios Retrospectivos , Manejo de Especímenes/normas , Manejo de Especímenes/estadística & datos numéricos , Factores de Tiempo
15.
J Chromatogr A ; 1635: 461731, 2021 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-33285415

RESUMEN

Conventional sampling of biological fluids often involves a bulk quantity of samples that are tedious to collect, deliver and process. Miniaturized sampling approaches have emerged as promising tools for sample collection due to numerous advantages such as minute sample size, patient friendliness and ease of shipment. This article reviews the applications and advances of microsampling techniques in therapeutic drug monitoring (TDM), covering the period January 2015 - August 2020. As whole blood is the gold standard sampling matrix for TDM, this article comprehensively highlights the most historical microsampling technique, the dried blood spot (DBS), and its development. Advanced developments of DBS, ranging from various automation DBS, paper spray mass spectrometry (PS-MS), 3D dried blood spheroids and volumetric absorptive paper disc (VAPD) and mini-disc (VAPDmini) are discussed. The volumetric absorptive microsampling (VAMS) approach, which overcomes the hematocrit effect associated with the DBS sample, has been employed in recent TDM. The sample collection and sample preparation details in DBS and VAMS are outlined and summarized. This review also delineates the involvement of other biological fluids (plasma, urine, breast milk and saliva) and their miniaturized dried matrix forms in TDM. Specific features and challenges of each microsampling technique are identified and comparison studies are reviewed.


Asunto(s)
Líquidos Corporales/química , Monitoreo de Drogas/métodos , Monitoreo de Drogas/tendencias , Manejo de Especímenes/métodos , Manejo de Especímenes/tendencias , Humanos
16.
Emerg Infect Dis ; 27(1)2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33183494

RESUMEN

Pooled nucleic acid amplification tests for severe acute respiratory syndrome coronavirus 2 could increase availability of testing at decreased cost. However, the effect of dilution on analytical sensitivity through sample pooling has not been well characterized. We tested 1,648 prospectively pooled specimens by using 3 nucleic acid amplification tests for severe acute respiratory syndrome coronavirus 2: a laboratory-developed real-time reverse transcription PCR targeting the envelope gene, and 2 commercially available Panther System assays targeting open reading frame 1ab. Positive percent agreement (PPA) of pooled versus individual testing ranged from 71.7% to 82.6% for pools of 8 and from 82.9% to 100.0% for pools of 4. We developed and validated an independent stochastic simulation model to estimate effects of dilution on PPA and efficiency of a 2-stage pooled real-time reverse transcription PCR testing algorithm. PPA was dependent on the proportion of tests with positive results, cycle threshold distribution, and assay limit of detection.


Asunto(s)
/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , /aislamiento & purificación , /virología , Técnicas de Laboratorio Clínico/métodos , Reacciones Falso Negativas , Humanos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificación de Ácido Nucleico/normas , Estudios Prospectivos , Sensibilidad y Especificidad , Manejo de Especímenes , Procesos Estocásticos
20.
Clin Microbiol Infect ; 27(1): 128.e1-128.e7, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32979576

RESUMEN

OBJECTIVES: A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the current coronavirus disease 2019 global pandemic. Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires a biosafety level 3 laboratory. This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2. METHODS: We used an automated microscope based on high-content screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells. A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared with the classical isolation procedure. RESULTS: The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct. Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared with only 26% (8/30) with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture. CONCLUSIONS: This system allows the rapid and automated screening of clinical samples with minimal operator workload, which reduces the risk of contamination and paves the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing.


Asunto(s)
/métodos , Procesamiento de Imagen Asistido por Computador/estadística & datos numéricos , ARN Viral/análisis , /aislamiento & purificación , Animales , Automatización de Laboratorios , Biomarcadores/análisis , Chlorocebus aethiops , Hospitalización , Humanos , Microscopía/métodos , Nasofaringe/virología , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Manejo de Especímenes/métodos , Células Vero , Carga Viral
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