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OBJECTIVES: Many hospitals use pneumatic tube systems (PTS) for transport of diagnostic samples. Continuous monitoring of PTS and evaluation prior to clinical use is recommended. Data loggers with specifically developed algorithms have been suggested as an additional tool in PTS evaluation. We compared two different data loggers. METHODS: Transport types - courier, conventional (cPTS) and innovative PTS (iPTS) - were monitored using two data loggers (MSR145® logger, CiK Solutions GmbH, Karlsruhe, Germany, and a prototype developed at the University Medicine Greifswald). Data loggers differ in algorithm, recording frequencies and limit of acceleration detection. Samples from apparently healthy volunteers were split among the transport types and results for 37 laboratory measurands were compared. RESULTS: For each logger specific arbitrary units were calculated. Area-under-the-curve (AUC)-values (MSR145®) were lowest for courier and highest for iPTS and increased with increasing recording frequencies. Stress (St)-values (prototype logger) were obtained in kmsu (1,000*mechanical stress unit) and were highest for iPTS as well. Statistical differences between laboratory measurement results of transport types were observed for three measurands sensitive for hemolysis. CONCLUSIONS: The statistical, but not clinical, differences in the results for hemolysis sensitive measurands may be regarded as an early sign of preanalytical impairment. Both data loggers record this important interval of beginning mechanical stress with a high resolution indicating their potential to facilitate early detection of preanalytical impairment. Further studies should identify suitable recording frequencies. Currently, evaluation and monitoring of diagnostic sample transport should not only rely on data loggers but also include diagnostic samples.
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Coleta de Amostras Sanguíneas , Hemólise , Humanos , Coleta de Amostras Sanguíneas/métodos , Estresse Mecânico , AlemanhaRESUMO
Due to the COVID-19 pandemic, many transport kits have been manufactured to preserve and transport nasopharyngeal swab samples (NPSs) from patients. However, there is no information on the performance of the different virus transport media (VTM) used in COVID-19 diagnosis in the population of Santiago de Chile. We compared the RT-qPCR amplification profile of five different viral transport kit mediums, including DNA/RNA Shield™, NAT, VTM-N, Ezmedlab™, and phosphate-buffered saline (PBS), for NPSs from Central Metropolitan Health Service, Santiago, Chile. The DNA/RNA Shield™ medium showed a better performance in terms of Cq and RFU values for the internal reference RNase P and viral ORF1ab probes. By contrast, the PBS transport medium registered higher Cq values for the viral and reference gene, compared to the other VTM. DNA/RNA Shield™ shows higher relative fluorescence units (RFUs) and lower Cq values for the reference gene. Collectively, our results suggest that the PBS medium could compromise the sample diagnosis because of its lower RT-qPCR performance. The NAT, Ezmedlab and VTM-N, and DNA/RNA Shield™ media show acceptable RT-qPCR parameters and, consequently, seem suitable for use in COVID-19 diagnosis.
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COVID-19 , COVID-19/diagnóstico , Teste para COVID-19 , Chile , Meios de Cultura , Humanos , Nasofaringe , Pandemias , RNA , RNA Viral/análise , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , SARS-CoV-2/genética , Manejo de Espécimes/métodosRESUMO
OBJECTIVES: Diagnostic samples are exposed to a spectrum of variables during transport to laboratories; therefore, the evaluation of a rather comprehensive stability profile of measurands is warranted. While appropriate testing standards have been established for pharmaceuticals and reagents, this is not the case for diagnostic samples. The aim of our work was to develop and evaluate a protocol applicable to diagnostic samples. METHODS: An isochronous approach with representation of temperature and exposure duration in a two-dimensional matrix was established. The deviations of the measurement results from the baseline associated with the exposure are evaluated with respect to the measurement uncertainty of the analytical measurement procedure applied. Variables of the experiment are documented in a standardized matrix. As a proof-of-concept, we profiled the stability patterns of a number of measurands at four temperature levels over up to 72 h in primary serum sample tubes. RESULTS: The protocol proved to be workable and allowed the description of a comprehensive stability profile of a considerable number of compounds based on 21 small-volume primary samples collected from each volunteer and exposed according to this protocol. CONCLUSIONS: A straightforward and feasible isochronous protocol can be used to investigate in detail the effects of different pre-processing conditions on the stability of measurands in primary samples during transport to diagnostic laboratories. This is of significance as pre-analytical logistics become increasingly important with the centralization of analytical services.
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Laboratórios , Humanos , Padrões de Referência , IncertezaRESUMO
OBJECTIVES: A carrier prototype by Aerocom® (Schwäbisch Gmünd, Germany) for pneumatic tube systems (PTS) is able to transport 9 blood tubes which are automatically fixed by closing the lid. In this study, we examined the influence of the transport on blood sample quality using the carrier prototype comparing to courier transport and a conventional carrier (AD160, Aerocom®). METHODS: Triplicate blood samples sets (1 lithium heparin, 1 EDTA, 1 sodium citrate) of 35 probands were split among the transportation methods: 1. courier, 2. conventional carrier, and 3. carrier prototype. After transport 51 measurands from clinical chemistry, hematology and coagulation were measured and compared. RESULTS: Overall, 49 of the investigated 51 measurands showed a good concordance among the three transport types, especially between the conventional carrier and the carrier prototype. Focusing on well-known hemolysis sensitive measurands, potassium showed no statistically significant differences. However, between courier and both carrier types lactate dehydrogenase (LDH) and free hemoglobin (fHb) showed statistically significant shifts, whereas the clinical impact of the identified differences was neglectable. The median concentration of fHb, for example, was 0.29 g/L (18 µmol/L), 0.31 g/L (19 µmol/L) and 0.32 g/L (20 µmol/L) for courier transport, conventional carrier and carrier prototype, respectively. These differences cannot be resolved analytically since the minimal difference (MD) for fHb is 0.052 g/L (3.23 µmol/L), at this concentration. CONCLUSIONS: The carrier prototype by Aerocom® is suitable for transportation of diagnostic blood samples. The overall workflow is improved by decreasing hands-on-time on the ward and laboratory while minimizing the risk of incorrectly packed carriers.
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Coleta de Amostras Sanguíneas , Hemólise , Coagulação Sanguínea , Hemoglobinas/análise , Humanos , L-Lactato Desidrogenase , PotássioRESUMO
The genetic characterization of measles viruses is an important tool for measles surveillance. Reverse cold chain requirements for the transportation of samples to reference laboratories are challenging in resource-limited settings. FTA cards facilitate the transport of virologic samples at ambient temperature as noninfectious material; however, the utility of FTA cards for the detection and genotyping of measles virus from clinical samples has not been evaluated. Throat swabs (TS) and oral fluid (OF) samples were collected from suspected measles cases in the Democratic Republic of the Congo. Virus detection (reverse transcription-quantitative real-time PCR [RT-qPCR]) and genotyping (endpoint RT-PCR) were compared for samples from 238 suspected cases; these samples were either transported using the reverse cold chain or at ambient temperature on FTA cards. Virus detection showed excellent positive agreement for OF samples compared to TS (95.3%; confidence interval [CI], 91.6 to 97.4), in contrast to 79.4% (CI, 73.5 to 84.3) for TS on FTA, and 85.5% (CI, 80.2 to 89.6) for OF on FTA compared to OF samples. Genotyping results obtained for a subset of samples indicated that 77.3% of all TS and 71.0% of OF samples would produce genotype information compared to 41.6% of TS and 41.3% of OF on FTA cards. Similar results were found for 16 measles-negative samples that were confirmed as rubella cases. Measles genotype B3 and rubella genotype 2B were detected. FTA cards have limited utility for virologic surveillance of sporadic cases of measles; however, they can be a useful tool for the expansion of virologic surveillance in countries where the reverse cold chain is not available.
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Vírus do Sarampo/isolamento & purificação , Boca/virologia , Faringe/virologia , Vírus da Rubéola/isolamento & purificação , Manejo de Espécimes/métodos , República Democrática do Congo , Genótipo , Técnicas de Genotipagem , Humanos , Sarampo/diagnóstico , Sarampo/virologia , Vírus do Sarampo/genética , Técnicas de Diagnóstico Molecular , RNA Viral/genética , Refrigeração , Rubéola (Sarampo Alemão)/diagnóstico , Rubéola (Sarampo Alemão)/virologia , Vírus da Rubéola/genética , Saliva/virologia , Manejo de Espécimes/instrumentaçãoRESUMO
BACKGROUND: Maintaining the quality of clinical specimens for tuberculosis (TB) testing is a major challenge in many high TB burden-limited resources countries. Sample referral systems in low and middle income countries are often weak and the maintenance of the cold-chain challenging and very costly for TB programs. The development of transport media allowing the preservation of samples without refrigeration is critical for increasing access to TB diagnostic services and for reducing the costs related to testing. METHODS: We evaluated the performance of OMNIgene-SPUTUM (OM-S) reagent for the maintenance of Mycobacterium tuberculosis (MTB) viability in sputum samples in the absence of refrigeration and its capacity to stabilize nucleic acid for molecular testing. A total of 329 sputum specimens from presumptive TB cases collected at the National Reference Laboratory in Tirana, Albania, were either decontaminated by a conventional method or processed with OM-S reagent and stored at room temperature. Samples in OM-S were shipped to the Supranational Reference Laboratory in Milan, Italy, at various times and processed for liquid culture. RESULTS: Our data show that OM-S maintains MTB viability for at least three weeks in the absence of refrigeration and improves the quality of culture resulting in a contamination rate lower than 0.5%. However, a significant delay in the time to culture positivity was observed for samples stored for more than two weeks in OM-S. CONCLUSIONS: Overall, OM-S offers multiple benefits both at laboratory and TB national program level by increasing the availability to quality diagnostics, promoting access to health care services and strengthening TB patient care especially in hard to reach populations.
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Mycobacterium tuberculosis , Manejo de Espécimes/métodos , Escarro/microbiologia , Tuberculose/microbiologia , Humanos , Indicadores e Reagentes , Estudos Prospectivos , Sensibilidade e Especificidade , Temperatura , Tuberculose/diagnósticoRESUMO
BACKGROUND: Culture contamination with environmental bacteria is a major challenge in tuberculosis (TB) laboratories in hot and humid climate zones. We studied the effect of cetylpyridinium chloride (CPC) preservation on culture results and performance of Xpert MTB/RIF. METHODS: Consecutive sputum samples from microscopy smear-positive TB patients were collected. Two-hundred samples were equally split in two aliquots, one aliquot was treated with CPC and stored at ambient temperature for 7 days. The second aliquot was immediately processed. Samples were decontaminated for 20, 15 or 10 min, and subsequently cultured on Löwenstein-Jensen medium. Furthermore, 50 samples were stored for 7, 14 and 21 days, and 100 CPC-pretreated samples tested by Xpert MTB/RIF. RESULTS: CPC pretreated samples showed a higher culture yield compared to non-treated sputum samples across all decontamination times: 94% vs. 73% at 10 min (p = 0.01), 94% vs. 64% at 15 min (p = 0.004), and 90% vs. 52% at 20 min (p < 0.001). The quantitative culture grading was consistently higher in CPC treated compared to non-CPC treated samples. The proportion of contaminated cultures was lower in CPC pretreated samples across all decontamination times (range 2-6%) compared to non-CPC treated samples (15-16%). For storage times of CPC treated samples of 7, 14, and 21 days, 84, 86, and 84% of the respective cultures were positive. Of 91 CPC treated samples with a positive culture, 90 were also Xpert MTB/RIF positive. CONCLUSIONS: CPC increases culture yield, decreases the proportion of contamination, and does not alter the performance of Xpert MTB/RIF.
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Técnicas Bacteriológicas/métodos , Cetilpiridínio/química , Mycobacterium tuberculosis/genética , Manejo de Espécimes/métodos , Escarro/microbiologia , Países em Desenvolvimento , Humanos , Microscopia/métodos , Mycobacterium tuberculosis/patogenicidade , Técnicas de Amplificação de Ácido Nucleico/métodos , Escarro/química , Tanzânia , Tuberculose Pulmonar/microbiologiaRESUMO
BACKGROUND: Recent papers on LA-ICP-MS have reported that certain elements are transported in particulate form, others in gaseous form and still others in a combination of both upon ablation of C-based materials. These two phases display different transport behaviour characteristics, potentially causing smearing in elemental maps, and could be processed differently in the ICP which raises concerns as to accuracy of quantification and emphasizes the need for matrix-matching of external standards. This work aims at a better understanding of two-phase sample transport by evaluating the peak profile changes observed upon varying parameters such as laser energy density and wavelength. RESULTS: It is demonstrated that upon ablation of gelatin, elements are transported predominantly in particulate phase, but already at low laser energy density, a significant fraction of some elements is transported in the gaseous phase, which is even more expressed at higher energy density. This behaviour is element-specific since the ratio of the signal intensity for the analyte element transported in gas phase to the total signal intensity was 0 % for 23Na, 43 % for 66Zn and as high as 95 % for 13C using a 193 nm laser. The results also suggest an effect of the laser wavelength, as all elements show either the same or higher amount of gas phase formation upon ablating with 213 nm versus 193 nm. It was even established that elements that fully occur in particulate form upon ablation using 193 nm laser radiation are partly converted into gaseous phase when using 213 nm. SIGNIFICANCE: This work provides a thorough investigation of the underexposed phenomenon of two-phase sample transport upon ablation of biological samples upon via LA-ICP-MS. It is shown that for some elements a fraction of the ablated material is converted and transported in the gas phase, which can lead to significant smearing effects. As such, it is important to evaluate element-specific peak profiles on beforehand and, if necessary, adapt instrument settings and slow down data acquisition.
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Gelatina , Terapia a Laser , Gases , Análise Espectral , Espectrometria de MassasRESUMO
Background: The appropriate sample handling for human fecal microbiota studies is essential to prevent changes in bacterial composition and quantities that could lead to misinterpretation of the data. Methods: This study firstly identified the potential effect of aerobic and anaerobic fecal sample collection and transport materials on microbiota and quantitative microbiota in healthy and fat-metabolic disorder Thai adults aged 23-43 years. We employed metagenomics followed by 16S rRNA gene sequencing and 16S rRNA gene qPCR, to analyze taxonomic composition, alpha diversity, beta diversity, bacterial quantification, Pearson's correlation with clinical factors for fat-metabolic disorder, and the microbial community and species potential metabolic functions. Results: Our study successfully obtained microbiota results in percent and quantitative compositions. Each sample exhibited quality sequences with a >99% Good's coverage index, and a relatively plateau rarefaction curve. Alpha diversity indices showed no statistical difference in percent and quantitative microbiota OTU richness and evenness, between aerobic and anaerobic sample transport materials. Obligate and facultative anaerobic species were analyzed and no statistical difference was observed. Supportively, the beta diversity analysis by non-metric multidimensional scale (NMDS) constructed using various beta diversity coefficients showed resembling microbiota community structures between aerobic and anaerobic sample transport groups (P = 0.86). On the other hand, the beta diversity could distinguish microbiota community structures between healthy and fat-metabolic disorder groups (P = 0.02), along with Pearson's correlated clinical parameters (i.e., age, liver stiffness, GGT, BMI, and TC), the significantly associated bacterial species and their microbial metabolic functions. For example, genera such as Ruminococcus and Bifidobacterium in healthy human gut provide functions in metabolisms of cofactors and vitamins, biosynthesis of secondary metabolites against gut pathogens, energy metabolisms, digestive system, and carbohydrate metabolism. These microbial functional characteristics were also predicted as healthy individual biomarkers by LEfSe scores. In conclusion, this study demonstrated that aerobic sample collection and transport (<48 h) did not statistically affect the microbiota and quantitative microbiota analyses in alpha and beta diversity measurements. The study also showed that the short-term aerobic sample collection and transport still allowed fecal microbiota differentiation between healthy and fat-metabolic disorder subjects, similar to anaerobic sample collection and transport. The core microbiota were analyzed, and the findings were consistent. Moreover, the microbiota-related metabolic potentials and bacterial species biomarkers in healthy and fat-metabolic disorder were suggested with statistical bioinformatics (i.e., Bacteroides plebeius).
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Fezes , Microbioma Gastrointestinal , RNA Ribossômico 16S , Humanos , Adulto , Microbioma Gastrointestinal/fisiologia , Microbioma Gastrointestinal/genética , Fezes/microbiologia , Tailândia , Masculino , RNA Ribossômico 16S/genética , Feminino , Adulto Jovem , Manejo de Espécimes/métodos , Anaerobiose/fisiologia , Aerobiose , Metagenômica , População do Sudeste AsiáticoRESUMO
OBJECTIVES: To determine optimal temperature profiles for 2 uniquely designed courier lockboxes (steel vs polymer) by standardizing daily ice (-20ºC) and cold (4-8ºC) pack placement. METHODS: Paired courier lockboxes were placed outside in direct sunlight. Ambient outdoor and lockbox temperatures were monitored during 2 4-day cycles, and temperature mean and range were determined daily (time frame, 4:00-10:00 pm). Control lockboxes without packs were compared with experimental paired lockboxes with either 2 cold packs placed at 4:00 pm or 4 ice packs placed at 8:00 am and replaced with 4 cold packs at 4:00 pm daily. RESULTS: Cycle 1 mean temperatures within control steel and polymer lockboxes were 31.8ºC (range, 18.4-44.1ºC) and 37.2ºC (range, 27.1-46.7ºC), respectively. The addition of 2 cold packs at 4:00 pm reduced mean temperatures to 29.1ºC (range, 19.1-37.2ºC) and 25.3ºC (range, 20.0-31.6ºC) in steel and polymer boxes, respectively. Cycle 2 mean temperatures within control steel and polymer lockboxes were 28.3ºC (range, 22.4-40.8ºC) and 31.6ºC (range, 23.8-41.0ºC), respectively. The addition of 4 ice packs at 8:00 am and replacement with 4 cold packs at 4:00 pm reduced mean temperatures to 24.3ºC (range, 17.4-27.9ºC) and 13.4ºC (range, 6.6-18.1ºC) in steel and polymer boxes, respectively. CONCLUSIONS: Standardizing instructions for ice and cold packs can decrease internal courier lockbox temperatures.
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Temperatura Alta , Gelo , Temperatura Baixa , Humanos , Polímeros , Aço , TemperaturaRESUMO
Epithelial tissue or vesicular fluid from an unruptured or recently ruptured vesicle is the sample of choice for confirmatory laboratory diagnosis of foot-and-mouth disease (FMD). However, in 'FMD-free' countries the transport and downstream processing of such samples from potentially infected animals present a biosafety risk, particularly during heightened surveillance, potentially involving decentralised testing in laboratories without adequate biocontainment facilities. In such circumstances, rapid inactivation of virus, if present, prior to transport becomes a necessity, while still maintaining the integrity of diagnostic analytes. Tongue epithelium collected from cattle infected with FMD virus (FMDV) of serotype O (O/ALG/3/2014 - Lineage O/ME-SA/Ind-2001d) or A (A/IRN/22/2015 - Lineage A/ASIA/G-VII) was incubated in the PAXGene Tissue System Fixative (pH 4) and Stabiliser (pH 6.5) components respectively, in McIlvaine's citrate-phosphate buffer (pH 2.6) or in phosphate-buffered saline (PBS, pH 7.4) at room temperature for 2, 6, 24 or 48â¯h. Following incubation, tissues were homogenised and tested by virus isolation and titration using LFBKαVß6 cells. The integrity of FMD viral RNA was assessed by RT-qPCR (3Dpol coding region), Sanger sequencing of the VP1 region and transfection of LFBKαVß6 cells to recover infectious virus. Viable virus could be recovered from samples incubated in PBS for at least 48â¯h. The PAXgene Tissue System Stabiliser component yielded variable results dependent on virus serotype, requiring at least 6â¯h of incubation to inactivate A/IRN/22/2015 in most samples, whereas the Fixative component required up to 2â¯h in some samples. McIlvaine's citrate-phosphate buffer rapidly inactivated both viruses within 2â¯h of incubation. There was no demonstrable degradation of FMD viral RNA resulting from incubation in any of the buffers for up to 48â¯h, as assessed by RT-qPCR, and 24â¯h by sequencing and transfection to recover infectious virus. McIlvaine's citrate-phosphate buffer (pH 2.6) is easy to prepare, inexpensive and inactivates serotype A and O FMDV in epithelial tissue within 2â¯h, while maintaining RNA integrity for downstream diagnostic processes and virus characterisation.
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Vírus da Febre Aftosa , Febre Aftosa , Animais , Bovinos , Citratos , Epitélio , Fixadores , Vírus da Febre Aftosa/genética , Fosfatos , RNA Viral/genética , Sorogrupo , LínguaRESUMO
Safe sample transport is of great importance for infectious diseases diagnostics. Various treatments and buffers are used to inactivate pathogens in diagnostic samples. At the same time, adequate sample preservation, particularly of nucleic acids, is essential to allow an accurate laboratory diagnosis. For viruses with single-stranded RNA genomes of positive polarity, such as foot-and-mouth disease virus (FMDV), however, naked full-length viral RNA can itself be infectious. In order to assess the risk of infection from inactivated FMDV samples, two animal experiments were performed. In the first trial, six cattle were injected with FMDV RNA (isolate A22/IRQ/24/64) into the tongue epithelium. All animals developed clinical disease within two days and FMDV was reisolated from serum and saliva samples. In the second trial, another group of six cattle was exposed to FMDV RNA by instilling it on the tongue and spraying it into the nose. The animals were observed for 10 days after exposure. All animals remained clinically unremarkable and virus isolation as well as FMDV genome detection in serum and saliva were negative. No transfection reagent was used for any of the animal inoculations. In conclusion, cattle can be infected by injection with naked FMDV RNA, but not by non-invasive exposure to the RNA. Inactivated FMDV samples that contain full-length viral RNA carry only a negligible risk of infecting animals.
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Doenças dos Bovinos , Vírus da Febre Aftosa , Febre Aftosa , Animais , Bovinos , Vírus da Febre Aftosa/genética , Genômica , RNA Viral/genéticaRESUMO
OBJECTIVES: To determine the impact of short-term (<4-hour) exposure of summer-like temperatures on lithium heparin (uncentrifuged and centrifuged) samples stored in outdoor courier lockboxes in the Mid-Atlantic United States. METHODS: Healthy adults (nâ =â 8) were recruited to investigate the impact of the short-term exposure of lithium heparin samples (centrifuged and uncentrifuged) inside 2 LabLocker-KF300 courier lockboxes placed outside in direct sunlight during summer. Each courier lockbox was monitored every 5 minutes with a temperature data logger and contained either the standard number (nâ =â 2) of cold packs (cold) or no standard cold packs (warm). Acceptable tolerance limits were defined for each analyte by significant change limit (SCL) analysis (Pâ <â .05), as previously described. RESULTS: Significant changes were identified in each study condition for warm and cold lockbox conditions. Aspartate aminotransferase, glucose, lactate dehydrogenase, and potassium commonly crossed SCLs from mean baseline (t0) in the majority of conditions. CONCLUSIONS: Outdoor courier lockboxes are an underrecognized source of preanalytical error.
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Análise Química do Sangue , Temperatura Alta , Manejo de Espécimes , HumanosRESUMO
Ambitions to improve the connectivity of devices to enable automation and digital representation of processes have been around for some time. Nevertheless, some companies, especially in life science and analytical science, tend to adopt these developments rather slowly. In the field of microbial analysis of drinking and process water, for example, a large part of the work is still carried out manually, although the high number of samples per day and the low fluctuation in work processes would predestine water analysis for a higher degree of automation. Obstacles such as the risk of bottlenecks and possible downtimes after machine failure, the spatial conditions together with the low flexibility of the automated system, a lack of trained personnel, and the high acquisition costs hinder this development, however.To lower these barriers, we have developed a system for the generation of flexibly expandable automated process lines, which handles sample handling and sample transport as a decisive step in the networking of several devices. The system allows the connection of devices that are distributed over the entire laboratory or close to each other, as well as those with a combination of both spatial situations. A functional or throughput expansion of the process can be realized by adding additional devices or storage areas to the network.With this concept, we have established a system for the automatic processing of defined steps of a routine Legionella pneumophila screening in drinking water testing. From this starting point, the process can be extended to cover further steps, such as concentrating or plating, up to the full analytical workflow.
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Laboratórios , Água , Automação , Testes Diagnósticos de Rotina , Fluxo de TrabalhoRESUMO
During a foot-and-mouth disease (FMD) outbreak, transport and testing of potentially infectious samples, including epithelium from suspect lesions, presents a biosafety risk, particularly in FMD-free countries. Therefore, treatment to inactivate virus prior to transport is important. Tongue epithelium from cattle infected with FMD virus (FMDV) serotype O (O ALG/3/2014 - Lineage O/ME-SA/Ind-2001d) or A (A IRN/22/2015 - Lineage A/ASIA/G-VII) was incubated in RNAlater, RNA Shield or phosphate-buffered saline (pH 7.4) at room temperature for 2, 6, 24 or 48 h. After incubation, tissues were homogenised and tested by virus titration. Viral RNA in the homogenate was quantified by RT-qPCR, used for sequencing, and transfected into LFBKαVß6 cells to recover infectious virus. RNAlater reduced A IRN/22/2015 titres by 4 log10 after 24 h, and completely after 48 h incubation. While O ALG/3/2014 was detected by VI after 2, 6 and 24 h, titration yielded no infectious virus, likely as a result of freeze-thawing. RNA Shield was cytotoxic at high concentrations but was effective at inactivating both strains after 24 h. Regardless of reagent or inactivation period, RT-qPCR, VP1 sequencing, and transfection of RNA to recover infectious virus were possible. RNA Shield appears a better choice for FMDV inactivation in tissues, however 24 h incubation is recommended.
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Epitélio/virologia , Vírus da Febre Aftosa/fisiologia , Manejo de Espécimes/métodos , Inativação de Vírus , Animais , Bovinos , Contenção de Riscos Biológicos , Vírus da Febre Aftosa/efeitos dos fármacos , Língua/citologia , Língua/virologia , Meios de TransporteRESUMO
BACKGROUND: Drones or unmanned aerial vehicles are autonomous or remotely controlled multipurpose aerial vehicles driven by aerodynamic forces and capable of carrying a payload. Whereas initially used exclusively for military purposes, the use of drones has gradually spread into other areas. Given their great flexibility and favourable costs, the use of drones has also been piloted in various healthcare settings. OBJECTIVES: We briefly summarize current knowledge regarding the use of drones in healthcare, focusing on infectious diseases and/or microbiology when applicable. SOURCES: Information was sought through PubMed and extracted from peer-reviewed literature published between January 2010 and August 2019 and from reliable online news sources. The search terms 'drones', 'unmanned aerial vehicles', 'microbiology' and 'medicine' were used. CONTENT: Peer-reviewed literature on the use of drones in healthcare has steadily increased in recent years. Drones have been successfully evaluated in various pilot programmes and are already implemented in some settings for transporting samples and delivering blood, vaccines, medicines, organs, life-saving medical supplies and equipment. In addition, a promising proof-of-concept 'lab-on-a-drone' was recently presented, as well as several pilot studies showing the benefits of drone use in surveillance and epidemiology of infectious diseases. IMPLICATIONS: The potential for drone use in clinical microbiology, infectious diseases and epidemiology is vast. Drones may help to increase access to healthcare for individuals that might otherwise not benefit from appropriate care due to remoteness and lack of infrastructure or funds. However, factors such as national airspace legislation and legal medical issues, differences in topography and climates, cost-effectiveness, and community attitudes and acceptance in different cultures and societies currently impede the widespread use of drones. Significant cost savings compared with ground transportation, speed and convenience of delivery, and the booming drone sector will probably drive drone implementation in various areas of medicine in the next 5 years.
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Técnicas de Laboratório Clínico/instrumentação , Atenção à Saúde/métodos , Medicina Militar/instrumentação , Doenças Transmissíveis/diagnóstico , Custos e Análise de Custo , Atenção à Saúde/normas , Serviços Médicos de Emergência/métodos , Humanos , Técnicas Microbiológicas/instrumentação , Medicina Militar/métodosRESUMO
Shipping of serum samples that were taken from pigs infected with classical swine fever (CSF) virus is frequently requested with the objective of serological analyses, not only for diagnostic purposes but also for exchange of reference materials that are used as control material of diagnostic assays. On the basis of the fact that an outbreak with CSF is associated with enormous economic losses, biological safety during the exchange of reference material is of great importance. The present study aimed to establish a pragmatic approach for reliable CSF virus (CSFV) inactivation in serum without impairing antibody detection. Considering the fact that complement inactivation through heating is routinely applied, the basic idea was to combine heat treatment with the dilution of serum in a detergent containing buffer in order to facilitate the inactivation process. The results show that treatment of serum samples with phosphate buffered saline-Tween20 (final concentration = 0.15%) along with incubation at 56 °C for 30 min inactivated CSFV and such treatment with ≤ 0.25% PBS-Tween20 does not impair subsequent antibody detection by ELISA or virus neutralization test. This minimizes the risk of virus contamination and represents a valuable contribution to a safer CSF diagnosis on a national and international level.
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Abstract@#Transport of filter paper-dried blood spot samples is a critical procedure during the screening of neonatal inherited metabolic diseases, which is of great significance for the screening accuracy. In order to ensure the timing and safety of sample transport, the cold chain positioning system was initiated by Zhejiang Provincial Center for Quality Control of Neonatal Disease Screening since March 2015. Based on the framework of neonatal disease screening information management system, the function of the logistics transport management system was included in this positioning system, with aims to achieve the monitoring and tracking of sample transport processes through real-time positioning of the sample transport box via China Unicom 4G logistics card and global positioning system/BeiDou Navigation Satellite System. The samples are maintained in a transport environment at 2 to 8 °C via the temperature-controlled box made of 5 °C phase-changed cold-stored materials and general packet radio service (GPRS) temperature recorders. The mean pretest turnover duration reduced from 8.44 days to 5.03 days following introduction of the cold chain positioning system, and the percentage of timely sample delivery increased from 31.69% to 77.90%, while the withdrawal rate of unqualified samples reduced from 0.12% to 0.08%. The cold chain positioning system meets the requirements of transport of filter paper-dried blood spot samples, which has a high potential in screening of neonatal inherited metabolic diseases.
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To comply with the pre-analytical requirements of ISO EN 15189, we investigated the stability of potassium, a very critical and sensitive analyte. We took into count effects of duration, temperature and transport after 10 hours storage of human whole blood in serum and plasma. Blood of 12 healthy subjects was analyzed after 4, 6, 8 and 10 hours of storage. Three study groups were designed: samples stored in laboratory at room temperature, transported by car during 4 hours at a temperature of 21±1ÌC, with or without previous thermal shock (20 min at 4±1 ÌC) before transportation. Variations in concentration were expressed as mean bias from baseline using the analytical change limit (ACL) and the reference change value (RCV). Using RCV, we considered that potassium was biologically stable during 10 hours whatever our study groups. Considering ACL, potassium in serum was not stable after the thermal shock. We conclude that whole blood in lithium-heparin tubes may be used for routine potassium analysis even if long car transportation and previous thermal shock is involved. It confirms that potassium analysis can be still performed in locations distant from a medical laboratory.