Diagnostic Setting Research Articles

Magnetic diagnostics layout design for CFETR plasma equilibrium reconstruction

Plasma equilibrium reconstruction provides essential information for tokamak operation and physical analysis. An extensive and reliable set of magnetic diagnostics is required to obtain accurate plasma equilibrium. This study designs and optimizes the magnetic diagnostics layout for the reconstruction of the equilibrium of the plasma according to the scientific objectives, engineering design parameters, and limitations of the Chinese Fusion Engineering Test Reactor (CFETR). Based on the CFETR discharge simulation, magnetic measurement data are employed to reconstruct consistent plasma equilibrium parameters, and magnetic diagnostics’ number and position are optimized by truncated Singular value decomposition, verifying the redundancy reliability of the magnetic diagnostics layout design. This provides a design solution for the layout of the magnetic diagnostics system required to control the plasma equilibrium of CFETR, and the developed design and optimization method can provide effective support to design magnetic diagnostics systems for future magnetic confinement fusion devices.

Next generation sequencing of high-grade adult-type diffuse glioma in the Netherlands: interlaboratory variation in the primary diagnostic and recurrent setting.

Next generation sequencing (NGS) is an important tool used in clinical practice to obtain the required molecular information for accurate diagnostics of high-grade adult-type diffuse glioma (HGG). Since individual centers use either in-house produced or standardized panels, interlaboratory variation could play a role in the practice of HGG diagnosis and treatment. This study aimed to investigate the current practice in NGS application for both primary and recurrent HGG. This nationwide Dutch survey used the expertise of (neuro)pathologists and clinical scientists in molecular pathology (CSMPs) by sending online questionnaires on clinical and technical aspects. Primary outcome was an overview of panel composition in the different centers for diagnostic practice of HGG. Secondary outcomes included practice for recurrent HGG and future perspectives. Out of twelve neuro-oncology centers, the survey was filled out by eleven (neuro)pathologists and seven CSMPs. The composition of the diagnostic NGS panels differed in each center with numbers of genes ranging from 12 to 523. Differences are more pronounced when tests are performed to find therapeutic targets in the case of recurrent disease: about half of the centers test for gene fusions (60%) and tumor mutational burden (40%). Current notable interlaboratory variations as illustrated in this study should be reduced in order to refine diagnostics and improve precision oncology. In-house developed tests, standardized panels and routine application of broad gene panels all have their own advantages and disadvantages. Future research would be of interest to study the clinical impact of variation in diagnostic approaches.

Challenges and approaches to calibrating patient phenotype as evidence for cancer gene variant classification under ACMG/AMP guidelines.

Since first publication of the American College of Medical Genetics and Genomics/Association for Medical Pathology (ACMG/AMP) variant classification guidelines, additional recommendations for application of certain criteria have been released (https://clinicalgenome.org/docs/), to improve their application in the diagnostic setting. However, none have addressed use of the PS4 and PP4 criteria, capturing patient presentation as evidence towards pathogenicity. Application of PS4 can be done through traditional case-control studies, or "proband counting" within or across clinical testing cohorts. Review of the existing PS4 and PP4 specifications for Hereditary Cancer Gene Variant Curation Expert Panels revealed substantial differences in the approach to defining specifications. Using BRCA1, BRCA2 and TP53 as exemplar genes, we calibrated different methods proposed for applying the "PS4 proband counting" criterion. For each approach, we considered limitations, non-independence with other ACMG/AMP criteria, broader applicability, and variability in results for different datasets. Our findings highlight inherent overlap of proband-counting methods with ACMG/AMP frequency codes, and the importance of calibration to derive dataset-specific code weights that can account for potential between-dataset differences in ascertainment and other factors. Our work emphasizes the advantages and generalizability of logistic regression analysis over simple proband-counting approaches to empirically determine the relative predictive capacity and weight of various personal clinical features in the context of multigene panel testing, for improved variant interpretation. We also provide a general protocol, including instructions for data formatting and a web-server for analysis of personal history parameters, to facilitate dataset-specific calibration analyses required to use such data for germline variant classification.

Pathological Diagnosis of Pulmonary Aspergillosis.

Pulmonary aspergillosis constitutes an increasingly prevalent and potentially fatal complex of mycotic diseases, caused by different species of Aspergillus. The broad spectrum of pathological manifestations associated with pulmonary aspergillosis necessitates a differentiation of commensalism from saprophytic colonization, hypersensitivity reactions, and true invasive infections, which highlights the importance of histopathology as a gold standard in a diagnostic setting. For the past decades, changes in terminology and contradicting contributions from different diagnostic disciplines have made the classification of pulmonary aspergillosis rather confusing. This review offers a categorization of aspergillosis lesions based on what can be histopathologically identified and distinguished, differentiating between acute invasive infection and forms of subacute, chronic, and allergic diseases and coinfections, and summarizes important manifestations of lesions associated with the different forms of pulmonary aspergillosis.

Synovial Fluid C-reactive Protein Clinical Decision Limit and Diagnostic Accuracy for Periprosthetic Joint Infection.

Introduction C-reactive protein (CRP) has long served as a prototypical biomarker for periprosthetic joint infection (PJI). Recently, synovial fluid (SF)-CRP has garnered interest as a diagnostic tool, with several studies demonstrating its diagnostic superiority over serum CRP for the diagnosis of PJI. Although previous studies have identified diagnostic thresholds for SF-CRP, they have been limited in scope and employed various CRP assays without formal validation for PJI diagnosis. This study aimed to conduct a formal single clinical laboratory validation to determine the optimal clinical decision limit of SF-CRP for the diagnosis of PJI. Methods A retrospective analysis of prospectively collected data was performed using receiver operating characteristic (ROC) and area under the curve (AUC) analyses. Synovial fluid samples from hip and knee arthroplasties, received from over 2,600 institutions,underwent clinical testing for PJI at a single clinical laboratory (CD Laboratories, Zimmer Biomet, Towson, MD) between 2017 and 2022. Samples were assayed for SF-CRP, alpha-defensin, white blood cell count, neutrophil percentage, and microbiological culture. After applying selection criteria, the samples were classified with the 2018 ICM PJI scoring system as "infected," "not infected," or "inconclusive." Data were divided into training and validation sets. The Youden Index was employed to optimize the clinical decision limit. Results A total of 96,061 samples formed the training (n = 67,242) and validation (n = 28,819) datasets. Analysis of the biomarker median values, culture positivity, anatomic distribution, and days from aspiration to testing revealed nearly identical specimen characteristics in both the training set and validation set. SF-CRP demonstrated an AUC of 0.929 (95% confidence interval (CI): 0.926-0.932) in the training set, with an optimal SF-CRP clinical decision limit for PJI diagnosis of 4.45 mg/L. Applying this cutoff to the validation dataset yielded a sensitivity of 86.1% (95% CI: 85.0-87.1%) and specificity of 87.1% (95% CI: 86.7-87.5%). No statistically significant difference in diagnostic performance was observed between the validation and training sets. Conclusion This study represents the largest single clinical laboratory evaluation of an SF-CRP assay for PJI diagnosis. The optimal CRP cutoff (4.45 mg/L) for PJI, which yielded a sensitivity of 86.1% and a specificity of 87.1%, is specific to the assay methodology and laboratory performing the assay. We propose that an SF-CRP test with a laboratory-validated optimal clinical decision limit for PJI may be preferable, in a clinical diagnostic setting, to serum CRP tests that do not have laboratory-validated clinical decision limits for PJI.

Printing surface cuprous oxides featured liquid metal for non-enzymatic electrochemical glucose sensor

Electrochemical glucose sensors that rely on two-dimensional (2D) oxides have attracted significant attention owing to the strong sensing activity of 2D oxides, but their practical application is hindered by the complexity and high cost of fabrication of electrodes and integrated devices. Herein, a convenient and effective fabrication route that includes printing a Ga-based liquid metal (LM) as a current collection electrode, followed by growing electrochemically active 2D oxides directly on the surface of Ga-based LMs under mild conditions, is developed for non-enzyme-based electrochemical sensors. Specifically, 2D annealed Cu-Oxide (ACO) is successfully grown on a printed Ga electrode through a galvanic replacement reaction, resulting in the formation of a mechanically and electrically well-matched interface between the active sensing materials and the current collection substrate. Benefitting from the high quantity of 2D ACO and good charge transfer at the interface, the as-prepared ACO electrode exhibits attractive glucose sensing performance, with a wide linear range (1 μM-10 mM) of effective detection, low detection limit down to 1 μM, and high sensitivity of 0.87 μA·mM-1·cm-2. Our study highlights the potential of using LMs in bio-sensing applications and provides a non-enzyme-based electrochemical biosensor platform for effective glucose detection in diets and clinical diagnostic settings.

Diagnostic intervention improved health-related quality of life among teenagers with food allergy.

The aim was to examine if a diagnostic intervention set up to assess current food allergy to cow's milk, hen's egg, fish, or wheat among teenagers had an impact on generic and disease specific health-related quality of life (HRQL). The study compared HRQL scoring before and two years after the intervention, and in relation to age matched controls without reported food allergy. The study was performed within the Obstructive Lung Disease in Northern Sweden (OLIN) studies where a cohort study on asthma and allergic diseases among 8-year-old schoolchildren was initiated in 2006. At age 12 years, the 125/2612 (5%) children who reported allergy to cow's milk, hen's egg, fish, or wheat were invited to a diagnostic intervention including clinical examination, blood tests and evaluation by a pediatric allergist. Of 94 participants, 79 completed generic and disease specific HRQL questionnaires. Additionally, a random sample of 200 (62% of invited) children without food allergy from the OLIN cohort answered the generic HRQL questionnaire. The respondents of the HRQL questionnaires were re-examined two years later and 57 teenagers with and 154 without reported allergy participated. There were no significant differences in generic HRQL scores between teenagers with and without reported food allergy at study entry, or after the intervention. Among those with reported food allergy, we found a significant improvement in disease specific HRQL after the intervention (mean values: 3.41 vs 2.80, p<0.001). Teenagers with only food allergy had better disease specific HRQL compared to those with one, two or three concomitant allergic diseases, both before and after the intervention. Children with only food allergy significantly improved their HRQL after the intervention, 1.84 vs. 2.87 (p<0.001) but this association was not seen in children with one other allergic disorder (3.16 vs. 3.65, p = 0.121) or those with two or more allergic disorders (3.72 vs. 3.90, p = 0.148). The diagnostic intervention showed a long-term improvement of disease specific HRQL but not generic HRQL.

Detection of a novel gross deletion in the UNC13D gene ends the diagnostic odyssey for a family with familial hemophagocytic lymphohistiocytosis 3

BackgroundFamilial hemophagocytic lymphohistiocytosis (FHL) is an immunological disorder characterized by overactivation of macrophages and T lymphocytes. This autosomal recessive condition has been characterized into multiple types depending on the genetic etiology. FHL type 3 is associated with bi-allelic pathogenic variants in the UNC13D gene.Case presentationWe present a 12-year diagnostic odyssey for a family with FHL that signifies the advances of FHL genetic testing in a clinical genetic diagnostic laboratory setting. We describe the first case of a large UNC13D gross deletion in trans to a nonsense variant in a family with FHL3, which may have been mediated by Alu elements within introns 12 and 25 of the UNC13D gene.ConclusionsThis case highlights the importance of re-evaluating past genetic testing for a patient and family as test technology evolves in order to end a diagnostic odyssey.

Building flexible and robust analysis frameworks for molecular subtyping of cancers.

Molecular subtyping is essential to infer tumor aggressiveness and predict prognosis. In practice, tumor profiling requires in-depth knowledge of bioinformatics tools involved in the processing and analysis of the generated data. Additionally, data incompatibility (e.g., microarray versus RNA sequencing data) and technical and uncharacterized biological variance between training and test data can pose challenges in classifying individual samples. In this article, we provide a roadmap for implementing bioinformatics frameworks for molecular profiling of human cancers in a clinical diagnostic setting. We describe a framework for integrating several methods for quality control, normalization, batch correction, classification and reporting, and develop a use case of the framework in breast cancer.

Amplification-free miRNA detection with CRISPR/Cas12a system based on fragment complementary activation strategy.

CRISPR/Cas12a systems have been repurposed as powerful tools for developing next-generation molecular diagnostics due to their trans-cleavage ability. However, it was long considered that the CRISPR/Cas12a system could only recognize DNA targets. Herein, we systematically investigated the intrinsic trans-cleavage activity of the CRISPR/Cas12a system (LbCas12a) and found that it could be activated through fragmented ssDNA activators. Remarkably, we discovered that the single-stranded DNA (ssDNA) activators in the complementary crRNA-distal domain could be replaced by target miRNA sequences without the need for pre-amplification or specialized recognition mechanisms. Based on these findings, we proposed the "Fragment Complementary Activation Strategy" (FCAS) and designed reverse fluorescence-enhanced lateral flow test strips (rFLTS) for the direct detection of miRNA-10b, achieving a limit of detection (LOD) of 5.53 fM and quantifying the miRNA-10b biomarker in clinical serum samples from glioma patients. Moreover, for the first time, we have developed the FCAS-based CRISPR/Cas12a system for miRNA in situ imaging, effectively recognizing tumor cells. The FCAS not only broadens the scope of CRISPR/Cas12a system target identification but also unlocks the potential for in-depth studies of CRISPR technology in many diagnostic settings.

IMPAIRMENT OF AUDITORY FUNCTION IN PERSONS WITH TYPE 2 DIABETES MELLITUS DEPENDING ON THE LEVEL OF PERIPHERAL MYELIN PROTEIN

Introduction: One of the priority tasks of modern medicine is early diagnosis and treatment of micro- and macrovascular complications of diabetes mellitus (DM). Variability of carbohydrate metabolism indicators, metabolic dysfunction are leading factors in the development and progression of complications of type 2 DM with hearing loss due to degenerative changes in nerve fibers. An important aspect of diagnostic and treatment algorithms of SNHL in patients with type 2 DM is early detection of demyelinating processes to prevent neuronal structures damage. The purpose of the study is to determine the relationship between the level of peripheral myelin protein 22 (PMP 22) and the condition of the auditory analyzer according to the data of examination in patients with type 2 DM combined with hearing impairment. Material and methods: There were examined 30 patients with type 2 DM (15 men and 15 women aged 56,28±4,54 years) associated with degree I–II sensorineural disorders; 15 persons with signs of sensorineural hearing loss (SNHL) (8 men, 7 women aged 59,33±2,65 years) without diabetes and 15 controls. During the examination the studied patients were divided into groups: the 1st group (n=15) without or with isolated microvascular complications of type 2 DM, the 2nd group (n = 15) with neuro-, nephro- and retinopathy. All patients underwent a set of laboratory tests, clinical and instrumental diagnostics of auditory function. The obtained data subjected to statistical processing and correlations identified. Results: The analysis revealed that in patients both groups with type 2 DM and SNHL the level of PMP22 significantly increased compared to controls and the comparison group and more significantly in those with microangiopathies. Conclusions: A significant increase was revealed in the content of PMP 22 and manifestations of SNHL in patients with type 2 DM associated with microangiopathies that indicate demyelinating processes in the neural structures of the auditory analyzer.

Effects of aripiprazole on prolactin levels and differences in effectiveness in patients with schizophrenia: a post-hoc analysis of the real-world data of a multicenter study.

To investigate the effect of aripiprazole on prolactin levels in patients with schizophrenia and analyze whether varying baseline prolactin levels affect the effectiveness and safety of aripiprazole, in a real-life diagnostic and therapeutic setting in a post-hoc analysis. In this post-hoc analysis, patients with schizophrenia in the acute phase were divided into an elevated-prolactin group and a normal-prolactin group. After 8 weeks of aripiprazole treatment, changes in the proportion of patients with an abnormal prolactin level were analyzed in both groups, and the efficacy and safety of aripiprazole were compared between the two groups. The elevated-prolactin group had more women, a longer duration of disease, and lower Positive and Negative Syndrome Scale (PANSS) total and subscale scores at baseline compared with the normal-prolactin group (all P < 0.05), and there was no significant difference in the proportion of patients with prior use of antipsychotic medication between the two groups. Regardless of previous antipsychotic use, patients in both groups developed hyperprolactinemia (23/168 [13.7%] in those who had taken antipsychotics vs. 43/375 [11.4%] in those who had not). After 8 weeks of aripiprazole treatment, the proportion of patients with abnormal prolactin in the elevated-prolactin group significantly decreased with prolonged treatment (P < 0.001), and aripiprazole had no significant effect on the normal-prolactin group (P = 0.250). Normal-prolactin group showed better efficacy than the elevated-prolactin group, and the differences in efficacy between the two groups was observed from week 4 to the endpoint (all p<0.05). In total, 87.2% (68/78) patients experienced mild to moderate adverse events in the elevated-prolactin group, which was significantly more frequent compared with the normal-prolactin group 71.0% (365/514). In this real-world study, for patients with acute schizophrenia, aripiprazole was effective in lowering the proportion of patients with abnormal prolactin levels, while it had no significant effect on patients with normal baseline prolactin. After adjusting for factors such as sex, age, prior antipsychotic drugs use history and disease severity, effectiveness and safety of aripiprazole in patients with normal baseline prolactin was significantly better than that in patients with elevated baseline prolactin.

Prevalence and factors associated with sarcopenia among older adults in a post-acute hospital in Singapore.

Sarcopenia is common in older adults worldwide, but its prevalence varies widely owing to differences in diagnostic criteria, population sampled, and care setting. We aimed to determine the prevalence and factors associated with sarcopenia in patients aged 65 and above admitted to a post-acute hospital in Singapore. This was a cross-sectional study of 400 patients recruited from a community hospital in Singapore. Data including socio-demographics, physical activity, nutritional status, cognition, clinical and functional status, as well as anthropometric measurements were collected. Sarcopenia was defined using the Asian Working Group for Sarcopenia 2019 criteria [AWGS2019]. Of the 383 patients with complete datasets, overall prevalence of sarcopenia was 54% while prevalence of severe sarcopenia was 38.9%. Participants with increased age, male gender and a low physical activity level were more likely to be sarcopenic, while those with higher hip circumference and higher BMI of ≥27.5m/kg2 were less likely to be sarcopenic. Other than the above-mentioned variables, cognitive impairment was also associated with severe sarcopenia. More than 1 in 2 older adults admitted to a post-acute hospital in Singapore are sarcopenic. There is an urgent need to address this important clinical syndrome burden and to identify patients at risk of sarcopenia in post-acute settings in Singapore for early intervention.

Application of a Specific and Sensitive NQO1 Turn-On Near-Infrared Fluorescence Probe for Live Cancer Cell and Xenografted Tumor Imaging in Nude Mice.

Sensitive activity stains for enzymes selectively expressed in human cancers offer valuable tools for imaging with wide applications in experimental, diagnostic, and therapeutic settings. The scant expression of the antioxidant enzyme NQO1 in normal tissues and its great abundance in malignant counterparts due to the increased redox stress and hypoxia is one such example. Previously, we described a potent nontoxic probe that remains nonfluorescent but releases an intense fluorogenic compound after intracellular cleavage by NQO1 catalysis. This infrared probe with a 644nm emission has excellent tissue penetrating ability and low background absorption. Described here are methods (fluorescence microscopy, flow cytometry, and in vivo animal imaging) to rapidly image NQO1 activity in hypoxic and non-hypoxic cancer cells and tumors developed in live mouse xenograft models. The specificity of the dye for NQO1 in all three procedures was verified, and the methods should be useful for both in vitro and in vivo studies.

Screening and assessment of malnutrition in patients with liver cirrhosis.

The development and advancement of malnutrition is associated not only with the progression of hepatic dysfunction, but also with cirrhosis-related complications. However, the prevalence of malnutrition reported in different studies varies widely due to differences in diagnostic methods and patient investigation settings. Therefore, we need to identify malnourished patients promptly and accurately. The purpose of this review was to compare the validity and reliability of nutritional screening tools and to select the most appropriate nutritional risk screening for patients with cirrhosis. We compared nutritional risk screening tools such as the Nutritional Risk Screening 2002 (NRS-2002), Malnutrition Universal Screening Tool (MUST), Royal Free Hospital-Nutritional Prioritizing Tool (RFH-NPT) and Liver Disease Undernutrition Screening Tool (LDUST). Royal Free Hospital-Nutritional Prioritizing Tool (RFH-NPT) is more feasible to screen cirrhotic patients for nutritional risk, and is highly reproducible, considering the impact of sodium and water retention; so it is practical to screen cirrhotic patients via RFH-NPT for nutritional risk, subsequently, to evaluate the nutritional status of patients with nutritional risk via the Global Leadership Initiative on Malnutrition (GLIM) diagnostic criteria. L3-SMI (third lumbar-skeletal muscle index) can accurately define sarcopenia in cirrhotic patients and also be used for clinical nutritional status assessment.

Combining mass spectrometry, i2PEPICO, and FTIR spectroscopy: Comprehensive speciation in DMM/NO oxidation

Oxymethylene ethers (OMEs) are potential alternative fuels that can be produced in a sustainable way based on CO2 and electricity. Additionally, they exhibit low tendencies to emit NOx or soot during practical combustion. Dimethoxy methane (DMM, CH3OCH2OCH3) has received increasing attention as the smallest OME with a O-CH2-O moiety. In the face of a possible application in exhaust gas recirculation (EGR) scenarios, understanding the influence of NOx on DMM oxidation is of importance. In this work, eleven mixtures of DMM/NO/O2/Ar were investigated in an atmospheric laminar flow reactor employing an extensive set of diagnostics. Molecular-beam mass spectrometry (MBMS), Fourier-transform infrared (FTIR) spectroscopy, NOx chemiluminescence detection, and double imaging photoelectron photoion coincidence (i2PEPICO) spectroscopy were combined to obtain a reliable speciation. All in all, species concentrations from different instruments showed a good agreement and one technique could counterbalance the physical limitations of another technique in many cases. In this manner, accurate mole fraction profiles of intermediates like e.g. CH4, C2H4, NO2, and methyl formate (C2H4O2) were gained. Based on i2PEPICO results, nitromethane (CH3NO2) and trans-HONO could additionally be identified as crucial intermediates in the NO-assisted oxidation of DMM. The present data set therefore provides an excellent basis to enhance future model development.

Porous Silicon modified by Molecularly Imprinted Polymers for Interleukin-6 (IL-6) detection

In this study, we present an innovative optical biosensor designed for the precise detection of Interleukin-6 (IL-6), a crucial cytokine associated with various pathological conditions. Our biosensor is based on silicon porous material meticulously modified with a molecularly imprinted polymer (MIP), ensuring specific and sensitive recognition of IL-6 molecules. Fabrication process involves the electrochemical etching of silicon porous chips followed by the electrodeposition of MIP, tailored to selectively bind IL-6 targets. Through rigorous testing across a range of IL-6 concentrations, our sensor exhibits remarkable sensitivity, showcasing discernible optical responses proportional to the varying analyte concentrations. Furthermore, we assessed the sensor's performance using bovine serum, a complex biological matrix, to simulate real-world sample conditions. Encouragingly, the sensor maintains its selectivity and optical response in the presence of serum components, affirming its robustness and applicability in practical diagnostic settings.

Can internal medicine specialists diagnose functional somatic disorders (FSDs)? Training and comparison with FSD specialists.

Functional somatic disorders (FSD) are a common problem across medical settings and remain challenging to diagnose and treat. Many patients with FSD undergo sequential and unnecessary extensive diagnostic work-up, which is costly for society and stressful for patients. Previous studies have shown that the empirically based FSD diagnostic entities are interrater reliable and stable over time. The aim of this study was to investigate whether internists who have received adequate training and with sufficient time per patient could diagnose FSD. This was a prospective diagnostic accuracy study. The study was conducted from May 2020 to April 2022. The study included 27 consecutive patients referred by their general practitioner to a non-psychiatric diagnostic clinic for assessment of physical symptoms on suspicion of FSD. The internists received a 30-hour training course in the use of a tailored version of the SCAN interview. The main outcome measure was the agreement between the diagnoses of the internists and the reference diagnoses made by specialists in FSD on the basis of the full SCAN interview. The interrater agreement between the internists and the FSD experts was substantial for any FSD (kappa = 0.63) as well as multi-organ vs. single-organ FSD (kappa = 0.73), indicating good diagnostic agreement. Internists with proper training and sufficient time (3-4 hours) per patient can proficiently diagnose FSD employing a tailored version of the SCAN interview for use in a non-psychiatric diagnostic setting.

Species and genetic diversity of nontuberculous mycobacteria in suspected tuberculosis cases in East Azerbaijan, Iran: a cross-sectional analysis.

The incidence of nontuberculous mycobacterial (NTM) infections has increased worldwide, attracting attention in routine diagnostic settings, particularly among patients with suspected tuberculosis. This study aimed to acquire knowledge of NTM infections in patients with suspected tuberculosis and to evaluate the genetic diversity of the strains. In this study, 230 clinical specimens were collected from suspected tuberculosis patients. Following decontamination with N-Acetyl-L-cysteine-sodium hydroxide (NALC-NaOH), the sediments of specimens were inoculated onto Löwenstein-Jensen medium and then incubated at 37°C for 8 weeks. The samples that yielded positive cultures were evaluated through the sequencing of conserved fragments of IS6110 and hsp65. For those samples that were not identified as part of the M. tuberculosis complex (MTC) by IS6110 PCR, further analysis was conducted via PCR to detect fragments of the hsp65 gene. Twenty-one NTM species were isolated from 230 clinical specimens (14 NTM from pulmonary specimens and 7 from extrapulmonary specimens). Among these, 12 (57.14%) were rapid-growing mycobacteria (RGM), and 9 (42.85%) were slow-growing mycobacteria (SGM). No M. avium complex (MAC) was identified in any of the specimens. Notably, the M. kansasii, M. gordonae, and M. abscessus strains presented significant genetic diversity. The prevalence of infections attributed to nontuberculous species surpasses that attributed to tuberculosis. These findings underscore the importance of exploring NTM species in individuals suspected of having TB.

Validating Clinical Exome Sequencing (CES) Test for Copy Number Variation (CNV) Analysis in Patients with Autism Spectrum Disorder

A routine methodologies to detect germline copy number variants (CNVs) are ArrayCGH, and SNParray, while clinical exome sequencing (CES) it is not a standard methods used in clinical settings. This study aims to investigate the efficiency of CNVs identification by CES in the diagnosis of Autism spectrum disorder (ASD) based on a Italian cohort. In this cohort study, 50 patients with ASD diagnosis were recruited for ASD screening. Simultaneous CNVs and single nucleotide variation (SNV) analysis was conducted to identify the anomalies that can cause ASD phenotype. A total of 13 patients were identified with a causative CNVs among 15 (86.67%) SNP array positive patients. Showing that CES data can be used for detecting clinically relevant CNVs with high sensitivity for use in clinical diagnostic settings.