analytical specificity
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UNLABELLED: The aim of this study was to identify parameters influencing DNA extraction and PCR amplification efficiencies in an attempt to standardize Mucorales qPCR. The Fungal PCR Initiative Mucorales Laboratory Working Group distributed two panels of simulated samples to 26 laboratories: Panel A (six sera spiked with Mucorales DNA and one negative control serum) and Panel B (six Mucorales DNA extracts). Panel A underwent DNA extraction in each laboratory according to the local procedure and were sent to a central laboratory for testing using three different qPCR techniques: one in-house qPCR assay and two commercial assays (MucorGenius and Fungiplex). Panel B DNA extracts were PCR amplified in each laboratory using local procedures: nine in-house qPCR assays and two commercial kits (MucorGenius and MycoGENIE). All data were compiled and anonymously analyzed at the central laboratory. For Panel A, a total of six different automated platforms and five manual extraction methods were used. Positive rates were 64%, 70%, and 89%, for the MucorGenius, Fungiplex, and the in-house qPCR assay, respectively. Using a large volume of serum for DNA extraction provided the highest analytical sensitivity (82.5% for 1 mL compared with 62.7% for smaller volumes, P < 0.01). For Panel B, five in-house qPCR assays and two commercial kits had >78% positivity. Using larger PCR input volumes (≥7 μL) was associated with the highest sensitivity at 95.5% compared to 58.3% when lower input volumes were used (P < 0.01). Using larger sample volumes for nucleic acid extraction and DNA template volumes for PCR amplification significantly improves the performance of Mucorales qPCR when testing serum. IMPORTANCE: Mucormycosis is a life-threatening mold infection affecting immunosuppressed patients but also other patients with diabetes or trauma. Better survival is linked to shorter delays in diagnosis and treatment initiation. Detection of Mucorales-free DNA in serum or plasma using quantitative PCR allows a prompt diagnosis and earlier treatment. Several techniques and protocols of quantitative Mucorales PCR are used in Europe, and improving performance remains a common objective of laboratories participating in the fungal PCR Initiative Working Group. This study, which combined results from 26 laboratories in Europe, showed that the main parameters underpinning sensitivity are the preanalytical variables (volume of serum used for DNA extraction and DNA template volume), irrespective of the extraction platforms and qPCR assay/platform.
- MeSH
- diagnostické techniky molekulární normy metody MeSH
- DNA fungální * krev genetika MeSH
- kvantitativní polymerázová řetězová reakce * normy metody MeSH
- lidé MeSH
- Mucorales * genetika izolace a purifikace MeSH
- mukormykóza * diagnóza mikrobiologie krev MeSH
- senzitivita a specificita * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
OBJECTIVES: Nonpharmacologic interventions (NPIs) constitute an important part of treatment for older adults, cover a broad and diverse range of interventions, and have advantages over pharmacologic interventions (eg, limited adverse side effects). However, an unambiguous definition of NPIs is still lacking. Defining NPIs may facilitate research on this topic and enhance comparability of results between studies, and might help to face the challenges of recognition, acceptation, funding, and implementation. Therefore, the aim of this review was to provide an overview and comparison of the definitions of NPIs used in the current literature on older adults. DESIGN: A systematic review was performed to provide an overview of the definitions of NPIs that are used in the current literature on older populations and to organize the characteristics involved in the definitions. SETTING AND PARTICIPANTS: People ≥60 years of age were included, not limited to a specific setting. METHODS: A systematic search was performed in the following 5 databases: PubMed, Embase, Clarivate Analytics/Web of Science Core Collection, Cumulative Index to Nursing and Allied Health Literature, and Wiley/Cochrane Library. The time frame within the databases was from inception to December 4, 2023. Review articles, editorials and consensus papers were included. RESULTS: We included 28 articles. We organized the definitions of NPI according to 4 different aspects: types of interventions involved, target population, goals the interventions addressed, and requirements of the interventions. Definitions in the current literature can generally be divided into 2 groups: NPIs described as not involving medication, and more elaborated multidomain definitions. Based on the results, we formulated criteria for types of interventions that can be considered an NPI. CONCLUSIONS AND IMPLICATIONS: Using current descriptions and characteristics, elements for a new definition for NPIs were proposed. To improve research in this field, consensus needs to be reached regarding elements covered by a definition of NPIs.
Vitamin B12, cobalamin, is indispensable for humans owing to its participation in two biochemical reactions: the conversion of l-methylmalonyl coenzyme A to succinyl coenzyme A, and the formation of methionine by methylation of homocysteine. Eukaryotes, encompassing plants, fungi, animals and humans, do not synthesise vitamin B12, in contrast to prokaryotes. Humans must consume it in their diet. The most important sources include meat, milk and dairy products, fish, shellfish and eggs. Due to this, vegetarians are at risk to develop a vitamin B12 deficiency and it is recommended that they consume fortified food. Vitamin B12 behaves differently to most vitamins of the B complex in several aspects, e.g. it is more stable, has a very specific mechanism of absorption and is stored in large amounts in the organism. This review summarises all its biological aspects (including its structure and natural sources as well as its stability in food, pharmacokinetics and physiological function) as well as causes, symptoms, diagnosis (with a summary of analytical methods for its measurement), prevention and treatment of its deficiency, and its pharmacological use and potential toxicity.
- MeSH
- dieta vegetariánská MeSH
- dieta MeSH
- fortifikované potraviny MeSH
- lidé MeSH
- nedostatek vitaminu B12 * diagnóza prevence a kontrola farmakoterapie etiologie MeSH
- vitamin B 12 * farmakokinetika chemie metabolismus terapeutické užití fyziologie škodlivé účinky aplikace a dávkování farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Práce se zabývá využitím algoritmů umělé inteligence (artificial intelligence – AI) v diagnostice karcinomu prsu, plic a prostaty. Popisuje historický vývoj digitalizace patologických procesů, implementaci umělé inteligence a její současné aplikace v patologii. Zaměřuje se na strojové a hluboké učení, počítačové vidění a digitální patologii, které přispívají k automatizaci a zpřesnění diagnostiky. Důraz je kladen na konkrétní nástroje, jako jsou systémy uPath od Roche a IBEX Medical Analytics, které umožňují analýzu histopatologických snímků, klasifikaci nádorových buněk a hodnocení biomarkerů. Práce také reflektuje výhody využití AI, včetně zvýšení přesnosti diagnostiky a efektivity laboratorních procesů, ale zároveň upozorňuje na výzvy spojené s její implementací, jako jsou etické a právní aspekty, ochrana osobních údajů a odpovědnost za chyby. Cílem práce je poskytnout komplexní přehled o možnostech využití AI v digitální patologii a její roli v moderní onkologické diagnostice.
The study focuses on the utilization of artificial intelligence (AI) algorithms in the diagnosis of breast, lung, and prostate cancer. It describes the historical development of the digitalization of pathological processes, the implementation of artificial intelligence, and its current applications in pathology. The study emphasizes machine learning, deep learning, computer vision, and digital pathology, which contribute to the automation and refinement of diagnostics. Special attention is given to specific tools such as the uPath systems from Roche and IBEX Medical Analytics, which enable the analysis of histopathological images, tumor cell classification, and biomarker evaluation. The study also highlights the benefits of AI utilization, including increased diagnostic accuracy and efficiency in laboratory processes, while simultaneously addressing the challenges associated with its implementation, such as ethical and legal considerations, data protection, and liability for errors. The aim of this study is to provide a comprehensive overview of the potential applications of AI in digital pathology and its role in modern oncological diagnostics.
Správný odběr biologického materiálu je důležitým předpokladem pro získání spolehlivých laboratorních výsledků. Laboratorní vyšetření zahrnuje tři hlavní fáze (preanalytickou, analytickou a postanalytickou), přičemž zejména preanalytická fáze je nejčastějším zdrojem chyb. Zásadní význam má zejména řádné poučení pacienta, přesná identifikace vzorků a požadovaných vyšetření, volba vhodných odběrových nádob, samotný postup při odběru biologického materiálu a podmínky uchování a transportu odebraného materiálu. Tento článek shrnuje klíčové aspekty správného odběru různých typů biologického materiálu a upozorňuje na nejčastější chyby, které mohou vést ke znemožnění vyšetření, zkreslení výsledků a mít tak negativní vliv na správnou interpretaci laboratorního nálezu.
Proper collection of biological material is essential for obtaining reliable laboratory results. Laboratory testing includes three main phases: pre-analytical, analytical, and post-analytical. The pre-analytical phase is the most frequent source of error. Thorough patient instruction, accurate sample and test identification, appropriate container selection, proper collection procedures, and adherence to the collected material's specific storage and transport conditions are all of particular importance. This article summarises the crucial aspects of the proper collection of various types of biological material (e.g. blood, urine, and sputum) and highlights the most common errors that can result in failed tests, inaccurate results, or a negative impact on the interpretation of results.
- MeSH
- klinické laboratorní techniky MeSH
- lidé MeSH
- odběr biologického vzorku * MeSH
- preanalytická fáze MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Growing evidence suggests that specific volatile organic compound (VOC) profiles may reflect key pathophysiological processes in Parkinson's disease (PD), including alterations in the microbiome, metabolism, and oxidative stress. Identifying reliable VOC biomarkers could enable non-invasive tests for early diagnosis, disease monitoring, and therapy evaluation. This review examines VOC analysis in biological matrices such as breath, skin, and stool, outlining current research and future applications in PD. We evaluate analytical techniques based on sensitivity, specificity, and clinical applicability. Additionally, we classify VOCs identified in previous studies alongside their proposed biological origins. Special attention is given to short-chain fatty acids, produced by the gut microbiome, a novel target in PD research. Our findings highlight the need for larger cohort studies and standardized protocols to advance VOC-based diagnostics in PD. Understanding the interplay between VOCs and PD may facilitate biomarker discovery, enhancing non-invasive diagnostic strategies and personalized disease management.
- Publikační typ
- časopisecké články MeSH
OBJECTIVE: Preeclampsia is a major pregnancy complication that results in significant maternal and infant mortality and morbidity, yet difficulties remain in the diagnosis of preeclampsia based on clinical parameters alone. The objective was to assess the performance of a hand-held point-of-care (POC) immunoassay in a clinical environment for glycosylated fibronectin (GlyFn) for the prediction of preeclampsia within 4 weeks of sampling. METHODS: Multinational European prospective observational pilot study of predominantly high-risk patients in the second half of pregnancy to assess a point-of-care immunoassay for GlyFn in predicting preeclampsia within 4 weeks of sampling. GlyFn was measured using a second generation hand held POC immunoassay. Results were considered normal for GlyFn concentrations of < 350 μg/mL, positive for GlyFn concentrations of 351-600 μg/mL, and high-positive for GlyFn concentrations > 600 μg/mL. RESULTS: Preeclampsia developed in 16 (19%) of 84 subjects and was associated with a shorter gestational age at delivery 35.3 weeks vs. 37.3 weeks for non-preeclamptics, n = 82; p = 0.001), a higher risk of fetal growth restriction (FGR; 31.2% vs. 10.3% for non-preeclamptics, p = 0.046), and an increased risk of preterm birth < 37 weeks gestation (83.3% vs. 33.3% for non-preeclamptics, (n = 78; p = 0.003). GlyFn positive or high positive was seen in 13/16 (81%) and in 35/68 (51.5%), yielding a sensitivity of 81%, a specificity of 49%, a positive predictive value of 27%, and a negative predictive value of 92%. GlyFn positive or high positive was also associated with preterm birth < 37 weeks in singleton pregnancy non-preeclamptic patients. Preterm birth occurred in 4.8% of those with normal GlyFn, in 26.7% with positive GlyFn, and in 50% of those with high GlyFn in singleton gestations without preeclampsia (p = 0.008). CONCLUSION: The ability to use this test in a POC format provides a method for practitioners to quickly determine risk for preeclampsia in their pregnant patients and offers an affordable alternative, as a single analyte to other diagnostic or screening tests that require laboratory-based testing or ultrasound equipment. Independent of preeclampsia, an elevated GlyFn was also correlated with preterm delivery and requires further study.
- MeSH
- biologické markery analýza MeSH
- dospělí MeSH
- fibronektiny * analýza MeSH
- gestační stáří MeSH
- glykované proteiny MeSH
- imunoanalýza metody MeSH
- lidé MeSH
- pilotní projekty MeSH
- point of care testing MeSH
- prediktivní hodnota testů MeSH
- preeklampsie * diagnóza MeSH
- prospektivní studie MeSH
- těhotenství MeSH
- vyšetření u lůžka MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- pozorovací studie MeSH
OBJECTIVES: Illicit drug use presents a significant challenge to global health and public safety, requiring innovative and effective monitoring strategies. This study aimed to evaluate the current landscape of wastewater-based epidemiology (WBE) for monitoring illicit drugs in Europe, focusing on collaboration, current practices, and barriers, while identifying opportunities for improvement. STUDY DESIGN: Cross-sectional survey-based study. METHODS: Coordinated by the Sewage Analysis CORe Group Europe (SCORE) and the European Union Drugs Agency (EUDA), two surveys were conducted in 2023 targeting researchers and stakeholders using WBE for illicit drugs. Data were analysed to identify trends, gaps, and opportunities for improving WBE implementation. RESULTS: The findings indicate a robust research infrastructure and diverse analytical methods among European institutions. Two-thirds of the participating countries reported using WBE data to inform policy. However, challenges persist, particularly in securing funding and coordination, as well as generating national estimates from multiple locations and addressing specific local policy needs. CONCLUSIONS: WBE has proven to be a valuable tool for monitoring illicit drug trends and informing drug policies. To unlock its full potential, sustained funding, methodological standardization, and enhanced cooperation are essential. This study provides critical insights into the European WBE landscape, offering a roadmap for strengthening the integration of actionable WBE data into public health and policy frameworks.
- MeSH
- epidemiologie odpadních vod MeSH
- lidé MeSH
- odpadní voda * MeSH
- poruchy spojené s užíváním psychoaktivních látek epidemiologie MeSH
- průřezové studie MeSH
- průzkumy a dotazníky MeSH
- rozhodování MeSH
- zakázané drogy * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
BACKGROUND: Observational data on composite scores often comes with missing component information. When a complete-case (CC) analysis of composite scores is unbiased, preferable approaches of dealing with missing component information should also be unbiased and provide a more precise estimate. We assessed the performance of several methods compared to CC analysis in estimating the means of common composite scores used in axial spondyloarthritis research. METHODS: Individual mean imputation (IMI), the modified formula method (MF), overall mean imputation (OMI), and multiple imputation of missing component values (MI) were assessed either analytically or by means of simulations from available data collected across Europe. Their performance in estimating the means of the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), the Bath Ankylosing Spondylitis Functional Index (BASFI), and the Ankylosing Spondylitis Disease Activity Score based on C-reactive protein (ASDAS-CRP) in cases where component information was set missing completely at random was compared to the CC approach based on bias, variance, and coverage. RESULTS: Like the MF method, IMI uses a modified formula for observations with missing components resulting in modified composite scores. In the case of an unbiased CC approach, these two methods yielded representative samples of the distribution arising from a mixture of the original and modified composite scores, which, however, could not be considered the same as the distribution of the original score. The IMI and MF method are, thus, intrinsically biased. OMI provided an unbiased mean but displayed a complex dependence structure among observations that, if not accounted for, resulted in severe coverage issues. MI improved precision compared to CC and gave unbiased means and proper coverage as long as the extent of missingness was not too large. CONCLUSIONS: MI of missing component values was the only method found successful in retaining CC's unbiasedness and in providing increased precision for estimating the means of BASDAI, BASFI, and ASDAS-CRP. However, since MI is susceptible to incorrect implementation and its performance may become questionable with increasing missingness, we consider the implementation of an error-free CC approach a valid and valuable option. TRIAL REGISTRATION: Not applicable as study uses data from patient registries.
- MeSH
- ankylózující spondylitida MeSH
- axiální spondyloartritida * MeSH
- C-reaktivní protein analýza MeSH
- interpretace statistických dat MeSH
- lidé MeSH
- stupeň závažnosti nemoci MeSH
- výzkumný projekt MeSH
- zkreslení výsledků (epidemiologie) MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
We present new developments for an ab-initio model of the neutron relative biological effectiveness (RBE) in inducing specific classes of DNA damage. RBE is evaluated as a function of the incident neutron energy and of the depth inside a human-sized reference spherical phantom. The adopted mechanistic approach traces neutron RBE back to its origin, i.e. neutron physical interactions with biological tissues. To this aim, we combined the simulation of radiation transport through biological matter, performed with the Monte Carlo code PHITS, and the prediction of DNA damage using analytical formulas, which ground on a large database of biophysical radiation track structure simulations performed with the code PARTRAC. In particular, two classes of DNA damage were considered: sites and clusters of double-strand breaks (DSBs), which are known to be correlated with cell fate following radiation exposure. Within a coherent modelling framework, this approach tackles the variation of neutron RBE in a wide energy range, from thermal neutrons to neutrons of hundreds of GeV, and reproduces effects related to depth in the human-sized receptor, as well as to the receptor size itself. Besides providing a better mechanistic understanding of neutron biological effectiveness, the new model can support better-informed decisions for radiation protection: indeed, current neutron weighting (ICRP)/quality (U.S. NRC) factors might be insufficient for use in some radiation protection applications, because they do not account for depth. RBE predictions obtained with the reported model were successfully compared to the currently adopted radiation protection standards when the depth information is not relevant (at the shallowest depth in the phantom or for very high energy neutrons). However, our results demonstrate that great care is needed when applying weighting factors as a function of incident neutron energy only, not explicitly considering RBE variation in the target. Finally, to facilitate the use of our results, we propose look-up RBE tables, explicitly considering the depth variable, and an analytical representation of the maximal RBE vs. neutron energy.