Simultaneous frequency and phase corrections of single-shot MRS data using cross-correlation.

PURPOSE: The objective of this study was to propose a novel preprocessing approach to simultaneously correct for the frequency and phase drifts in MRS data using cross-correlation technique. METHODS: The performance of the proposed method was first investigated at different SNR levels using simulation. Random frequency and phase offsets were added to a previously acquired STEAM human data at 7 T, simulating two different noise levels with and without baseline artifacts. Alongside the proposed spectral cross-correlation (SC) method, three other simultaneous alignment approaches were evaluated. Validation was performed on human brain data at 3 T and mouse brain data at 16.4 T. RESULTS: The results showed that the SC technique effectively corrects for both small and large frequency and phase drifts, even at low SNR levels. Furthermore, the mean square measurement error of the SC algorithm was comparable to the other three methods used, with much faster processing time. The efficacy of the proposed technique was successfully demonstrated in both human brain MRS data and in a noisy MRS dataset acquired from a small volume-of-interest in the mouse brain. CONCLUSION: The study demonstrated the availability of a fast and robust technique that accurately corrects for both small and large frequency and phase shifts in MRS.

T1 and T2 measurements across multiple 0.55T MRI systems using open-source vendor-neutral sequences.

PURPOSE: To compare T1 and T2 measurements across commercial and prototype 0.55T MRI systems in both phantom and healthy participants using the same vendor-neutral pulse sequences, reconstruction, and analysis methods. METHODS: Standard spin echo measurements and abbreviated protocol measurements of T1, B1, and T2 were made on two prototype 0.55 T systems and two commercial 0.55T systems using an ISMRM/NIST system phantom. Additionally, five healthy participants were imaged at each system using the abbreviated protocol for T1, B1, and T2 measurement. The phantom measurements were compared to NMR-based reference measurements to determine accuracy, and both phantom and in vivo measurements were compared to assess reproducibility and differences between the prototype and commercial systems. RESULTS: Vendor-neutral sequences were implemented across all four systems, and the code for pulse sequences and reconstruction is freely available. For participants, there was no difference in the mean T1 and T2 relaxation times between the prototype and commercial systems. In the phantom, there were no significant differences between the prototype and commercial systems for T1 and T2 measurements using the abbreviated protocol. CONCLUSION: Quantitative T1 and T2 measurements at 0.55T in phantom and healthy participants are not statistically different across the prototype and commercial systems.

Evaluation of an Electrochemiluminescence Assay for the Rapid Detection of Ricin Toxin.

In this article, we detail a comprehensive laboratory evaluation of an immunoassay for the rapid detection of ricin using the Meso Scale Diagnostics Sector PR2 Model 1800. For the assay evaluation, we used inclusivity, exclusivity, and informational panels comprised of extracts of 35 near-neighbor plant cultivar-extracts, 66 lectins, 26 white powders, 16 closely related toxins and proteins/toxoids, and a pool of 30 BioWatch filter extracts. The results show that the Meso Scale Diagnostics ricin detection assay exhibits good sensitivity and specificity with a limit of detection of 1.2 ng/mL. However, the dynamic range of the assay for the quantitation of ricin was limited. We observed a hook effect at higher ricin concentrations, which can lead to potential false negative results. A modification of the assay protocol that incorporates extra wash steps can decrease the hook effect and the potential for false negative results.

Cell-free DNA end characteristics enable accurate and sensitive cancer diagnosis.

The fragmentation patterns of cell-free DNA (cfDNA) in plasma can potentially be utilized as diagnostic biomarkers in liquid biopsy. However, our knowledge of this biological process and the information encoded in fragmentation patterns remains preliminary. Here, we investigated the cfDNA fragmentomic characteristics against nucleosome positioning patterns in hematopoietic cells. cfDNA molecules with ends located within nucleosomes were relatively shorter with altered end motif patterns, demonstrating the feasibility of enriching tumor-derived cfDNA in patients with cancer through the selection of molecules possessing such ends. We then developed three cfDNA fragmentomic metrics after end selection, which showed significant alterations in patients with cancer and enabled cancer diagnosis. By incorporating machine learning, we further built high-performance diagnostic models, which achieved an overall area under the curve of 0.95 and 85.1% sensitivity at 95% specificity. Hence, our investigations explored the end characteristics of cfDNA fragmentomics and their merits in building accurate and sensitive cancer diagnostic models.

Bladder Cancer detection by urinary methylation markers GHSR/MAL: a validation study.

PURPOSE: Although cystoscopy is a reliable tool for detecting bladder cancer, it poses a high burden on patients and entails high costs. This highlights the need for non-invasive and cost-effective alternatives. This study aimed to validate a previously developed urinary methylation marker panel containing GHSR and MAL. METHODS: We enrolled 134 patients who underwent cystoscopy because of hematuria, including 63 individuals with primary bladder cancer and 71 with non-malignant findings. Urine samples were self-collected at home and sent via regular mail. Subsequently, DNA was extracted and the hypermethylation of GHSR and MAL was evaluated using quantitative methylation-specific polymerase chain reaction. The performance of methylation markers was assessed using area-under-the-curve (AUC) analysis and sensitivity and specificity based on pre-established cut-off values. RESULTS: Validation of the marker panel GHSR/MAL resulted in an AUC of 0.87 at 79% sensitivity and 80% specificity. Sensitivity was comparable to the previous investigation (P > 0.9), though specificity was significantly lower (P = 0.026). Sensitivity was higher for high-grade tumors compared to low-grade tumors (94% vs. 60%, P = 0.002). CONCLUSION: Validation of the GHSR/MAL methylation marker panel on at home collected urine samples confirms its robust performance for bladder cancer detection in a hematuria population, and underscores the diagnostic potential for future clinical application.

Evaluation of the analytical performance of four different manufacturer's reagent red blood cells in antibody detection and identification.

OBJECTIVE: The detection/identification of clinically significant antibodies to red cell antigens form the foundation for safe transfusion practices. This study aimed to evaluate the diagnostic performance of commercially available 0.8% reagent red blood cells (RRBCs) in Australia. 166 patient-derived plasma samples with a positive indirect antiglobulin test (IAT) were tested using column agglutination technology (CAT) with Immulab, Bio-Rad, Grifols and QuidelOrtho screening and identification RRBCs with the respective manufacturer's proprietary CAT system. RESULTS: False-negative antibody screening and identification results were obtained with Bio-Rad (3/61), Grifols (14/68) and Quidel-Ortho (3/59) RRBCs when tested with the respective manufacturer's proprietary CAT system. Zero false-negative results were observed with Immulab RRBCs when tested with samples across all platforms. The sensitivity of the RRBCs used in this study were calculated to be 95.83% (95%CI 88.30-99.13%) for Bio-Rad RRBCs, 82.50% (95%CI 72.38-90.09%) for Grifols RRBCs and 95.65% (95%CI 87.82-99.09%) for QuidelOrtho RRBCs. The sensitivity of Immulab RRBCs were stratified based on performance in the 3 CAT platforms: Bio-Rad CAT (100%, 95%CI 95.01-100%), Grifols CAT (100%, 95%CI 95.49-100%) and QuidelOrtho CAT (100%, 95%CI 94.79-100%). CONCLUSIONS: RRBCs used in antibody detection and identification vary in diagnostic performance and should therefore be carefully considered before being implemented in routine patient testing.

Quantitative real time PCR for distinction between Pneumocystis jirovecii infection/colonization in hospitalized patients.

Background: Identification of the opportunistic fungus Pneumocystis jirovecii in respiratory specimens presents challenges, particularly in differentiating between colonization and active infection. The present study assessed a probe-based real time PCR (qPCR) diagnostic effectiveness in patients with diverse underlying conditions, particularly those with COVID-19 and pulmonary insufficiency. Methods: To set up the qPCR, clinical samples from 281 patients with respiratory ailments were tested. Subsequently, a descriptive study was conducted on 112 patients with pulmonary insufficiency with and without COVID-19 suspected of P. jirovecii infection. All specimens were subjected to DNA extraction followed by nested PCR and qPCR targeting the mitochondrial large subunit (mtLSU)-rRNA gene. Results: Based on nested PCR and qPCR, P. jirovecii was identified in 40 out of 281 patients, with slight variations in positive samples observed across dilutions. Three patients who tested positive in nested PCR yielded negative results in probe-based qPCR. Conversely, three patients who tested positive in probe-based qPCR yielded negative results in nested PCR. Considering nested PCR as the golden standard, probe-based qPCR demonstrated good diagnostic performance, with 92.5% sensitivity and 98.7% specificity. Based on cycle threshold (Ct) values, the positive cases were categorized: ≤32 as infection, >35 as colonization, and a grey zone between these values (32 < X ≤ 35). The analysis of 112 PCP-suspected patients revealed a prevalence ranging from 6.25% (nested PCR) to 7% (probe-based qPCR). Conclusions: This study suggested Ct values to differentiate Pneumocystis pneumonia/colonization in immunocompromised patients. To further augment the diagnostic sensitivity, it is recommended to integrate qPCR results with clinical parameters and biomarkers to offer a more precise understanding of Pneumocystis-related conditions.

Liver Injury in People With HIV on Antituberculosis and/or Antiretroviral Therapy-Assessing Causality Using the Updated Roussel Uclaf Causality Assessment Method.

PURPOSE: We compared performance of the Roussel Uclaf Causality Assessment Method (RUCAM) with multidisciplinary expert panel review in identifying a drug-induced liver injury (DILI) due to antituberculosis therapy (ATT) and/or antiretroviral therapy (ART). METHODS: Cases were drawn from a prospective registry of hospitalised adults with suspected DILI due to ATT and/or ART in Cape Town, South Africa. Participants had to fulfil American Thoracic Society criteria for ATT interruption (alanine transaminase [ALT] ≥5 times upper limit of normal [ULN]/ALT ≥3 times [ULN] and symptomatic). Causality assessment by expert panel review served as reference standard. The panel ranked potentially implicated drugs as certain, probable, possible or unlikely causes guided by World Health Organization Uppsala Monitoring Centre criteria. The RUCAM was performed for each potentially implicated drug. We calculated sensitivity and specificity of the RUCAM in identifying a probable/certain drug cause for liver injury. RESULTS: We included 48 participants. All were people with HIV (PWH). Twenty-seven were on concomitant ART and ATT, with a median of six potentially hepatotoxic drugs per case. Sensitivity and specificity of the RUCAM in identifying a probable/certain drug cause of liver injury compared with expert panel review was 7% and 100% respectively. Implicated drugs (times ranked probable/certain by panel) were isoniazid (18/0), pyrazinamide (17/0), rifampicin (15/1), efavirenz (6/4) and lopinavir/ritonavir (1/0). CONCLUSIONS: PWH with liver injury received multiple potentially implicated drugs, which may increase liver injury risk and complicate causality assessment. Compared with expert panel review, the RUCAM had low sensitivity in detecting probable or certain drug causes of liver injury.

MRI-based Zero Echo Time and Black Bone Pseudo-CT Compared with Whole-Body CT to Detect Osteolytic Lesions in Multiple Myeloma.

Background MRI is highly sensitive for assessing bone marrow involvement in multiple myeloma (MM) but does not enable detection of osteolysis. Purpose To assess the diagnostic accuracy, repeatability, and reproducibility of pseudo-CT MRI sequences (zero echo time [ZTE], gradient-echo black bone [BB]) in detecting osteolytic lesions in MM using whole-body CT as the reference standard. Materials and Methods In this prospective study, consecutive patients were enrolled in our academic hospital between June 2021 and December 2022. Inclusion criteria were newly diagnosed MM, monoclonal gammopathy of undetermined significance at high risk for MM, or suspicion of progressive MM. Participants underwent ZTE and BB sequences covering the lumbar spine, pelvis, and proximal femurs as part of 3-T whole-body MRI examinations, as well as clinically indicated fluorine 18 fluorodeoxyglucose PET/CT examination within 1 month that included optimized whole-body CT. Ten bone regions and two scores (categorical score = presence/absence of osteolytic lesion; semiquantitative score = osteolytic lesion count) were assessed by three radiologists (two experienced and one unfamiliar with pseudo-CT reading) on the ZTE, BB, and whole-body CT images. The accuracy, repeatability, and reproducibility of categorical scores (according to Gwet agreement coefficients AC1 and AC2) and differences in semiquantitative scores were assessed at the per-sequence, per-region, and per-patient levels. Results A total of 47 participants (mean age, 67 years ± 11 [SD]; 27 male) were included. In experienced readers, BB and ZTE had the same high accuracy (98%) in the per-patient analysis, while BB accuracy ranged 83%-100% and ZTE accuracy ranged 74%-94% in the per-region analysis. An increase of false-negative (FN) findings in the spine ranging from +17% up to +23%, according to the lumbar vertebra, was observed using ZTE (P < .013). Regardless of the region (except coxal bones), differences in the BB score minus the ZTE score were positively skewed (P < .021). Regardless of the sequence or region, repeatability was very good (AC1 ≥0.87 for all), while reproducibility was at least good (AC2 ≥0.63 for all). Conclusion Both MRI-based ZTE and BB pseudo-CT sequences of the lumbar spine, pelvis, and femurs demonstrated high diagnostic accuracy in detecting osteolytic lesions in MM. Compared with BB, the ZTE sequence yielded more FN findings in the spine. ClinicalTrials.gov Identifier: NCT05381077 Published under a CC BY 4.0 license. Supplemental material is available for this article.

Performance Assessment of the New EUROLINE Neurologic Syndrome 15 Ag (IgG) for the Determination of Autoantibodies Associated with Neurological Disorders.

OBJECTIVE: Here, we assess the performance of the new EUROLINE Neurologic Syndrome 15 Ag (IgG) which expands the EUROLINE Paraneoplastic Neurologic Syndrome 12 Ag by adding CDR2L (together with CDR2 targeted by anti-Yo), AK5, and Neurochondrin (NCDN). BACKGROUND: Many paraneoplastic as well as non-paraneoplastic autoantibodies (AAbs) have been described in neurological disorders in the last decade. By integrating the associated antigens into existing assays, the diagnostic work-up of patients is being improved and diagnostic gaps reduced. DESIGN/METHODS: Sensitivity of each AAb was analyzed using a total of 194 clinically and diagnostically pre-characterized samples (Table 1). Specificity of each AAb was investigated using a minimum of 100 sera from healthy blood donors. RESULTS: Using the EUROLINE Neurologic Syndrome 15 Ag, autoantibody positivity was confirmed in 89-100% of samples. In particular, all samples for which clinical and tissue-based indirect immunofluorescence assay pre-characterization indicated anti-Yo positivity were anti-CDR2 and -CDR2L double positive. Anti-AK5 was determined in serum and cerebrospinal fluid (CSF) with a sensitivity of 90 and 100%, respectively, and anti-NCDN with a sensitivity of 100%. The individual specificities were ≥99%. CONCLUSIONS: This kit is a tool for the qualitative in vitro determination of AAbs against a large panel of 15 different neuronal autoantigens to support the diagnosis of neurologic syndromes. The parallel detection of anti-CDR2 and anti-CDR2L (both anti-Yo) increases the diagnostic significance, as double positivity is strongly related to paraneoplastic cerebellar degeneration.[Table: see text].

Can Immature Granulocyte Counting be Utilized in the Selection of Samples for Flow Cytometric Enumeration of Stem Cells?

BACKGROUND: This study aimed to compare the IMG counting by an auto hematology analyzer with the flow cytometric enumeration of CD34+ cells. METHODS: All data from 124 samples submitted to the hematology laboratory for CD34+ cell counting in 2019 and 2020 were retrospectively evaluated. Whole blood samples were taken into EDTA tubes and assayed within 2 hours. The numbers and percentages of WBC and IMG were determined by using Mindray BC6200 hematology analyzer, while the same were determined for CD45 and CD34+ cells by using Beckman Coulter Navios flow cytometer. The performance of the new method was determined by the receiver operating characteristic (ROC) analysis. RESULTS: Out of the 124 samples, 94 were collected from healthy individuals and 30 were collected from patients. A significant correlation was found between IMG and CD34+ cell counts in all patients (r = 0.71, p = 0.000) at the cutoff values of > 20/µL and > 50/µL. The Bland-Altman analysis of all patients showed that there was an agreement between the two methods. When the cutoff value of 20/µL for CD34+ cells was used in the ROC analysis, the sensitivity and specificity were calculated as 100 (96.1 - 100) and 96.77 (83.3 - 99.9), respectively, for the IMG count of > 900/µL. CONCLUSIONS: An IMG count of 900/µL can be used as the cutoff value in the analysis with the Mindray BC6200. The IMG counting cannot replace the flow cytometric CD34+ cell enumeration but can be used in the selection of the samples for stem cell enumeration by flow cytometry.

Point-of-care testing for COVID-19: a simple two-step molecular diagnostic development and validation during the SARS-CoV-2 pandemic.

BACKGROUND: During the coronavirus disease 19 (COVID-19) pandemic, diagnostic testing of the general population proved challenging due to limitations of the gold-standard diagnostic procedure using reverse transcription real-time polymerase chain reaction (RT-qPCR) for large-scale testing on the centralised model, especially in low-resource areas. OBJECTIVES: To address this, a point-of-care (PoC) diagnostic protocol for COVID-19 was developed, providing fast, reliable, and affordable testing, particularly for low-mid develop areas. METHODS: The PoC diagnostic process combines a simple paper-based RNA extraction method housed within a 3D-printed plastic device with a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. Nasopharyngeal/oropharyngeal swabs (NOS) and saliva samples were tested between 2020 and 2021, with the assistance of Santa Catarina's State Health Secretary, Brazil. FINDINGS: The developed diagnostic protocol showed a limit of detection of 9,900 copies and an overall diagnostic specificity of 98% for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from 1,348 clinical analysed samples. The diagnostic sensitivity was 95% for NOS samples, 85% for early morning saliva, and 69% for indiscriminate saliva. MAIN CONCLUSIONS: In conclusion, the developed device successfully extracted SARS-CoV-2 viral RNA from swabs and saliva clinical samples. When combined with colorimetric RT-LAMP, it provides results within 45 min using minimal resources, thus delivering a diagnostic kit protocol that is applicable in large-scale sampling.

Utility of Sonoelastography and Magnetic Resonance Imaging in Characterization of Thyroid Nodules.

AIM: To evaluate thyroid nodules with sonoelastography and magnetic resonance imaging (MRI). MATERIALS AND METHODS: The study included 28 patients with 40 thyroid nodules. Clearance was obtained from the institute's ethical clearance committee. Patients with pure cystic nodules or nodules with eggshell calcification, diffuse thyroid pathology (such as Graves' disease, Hashimoto's thyroiditis, De Quervain thyroiditis, and Riedel's thyroiditis), inaccessible nodules via fine needle aspiration cytology (FNAC), or patients with a history of thyroid gland surgery were excluded from the study. Strain elastography was performed on a Phillips iU22 machine, producing qualitative color-coded strain maps (graded using the Rago 5-point system) and semiquantitative strain ratios. MRI was performed on a Phillips ACHIEVA 1.5T magnet with a head and neck coil. RESULTS: Rago scores statistically correlated (χ2 = 18.052, p < 0.001) with malignant nodules, and using the receiver operating characteristic (ROC) curve, the area under the ROC curve (AUROC) for the mean strain ratio predicting malignant outcomes was 0.88 [95% confidence interval (CI): 0.767-0.992], which was also statistically significant (p < 0.001). A cutoff of mean strain ratio ≥2.48 predicted malignant outcomes with 100% specificity. T2 signal intensity ratio (SIR) and apparent diffusion coefficient (ADC) values were not statistically significant in predicting malignant outcomes. Kinetic curves were statistically significant for Rago scores (χ2 = 11.356, p = 0.045); however, no significant difference was found in predicting malignant outcomes. CONCLUSION AND CLINICAL SIGNIFICANCE: We concluded that sonoelastography, along with grayscale ultrasound, is a useful noninvasive technique for predicting histological outcomes. However, MRI should largely be reserved as a problem-solving tool rather than a standalone imaging modality. The kinetic curves show some degree of overlap between histologically distinct diseases, and thus large-scale multicenter trials are needed for further standardization.

[Neonatal screening for cystic fibrosis in the Liège region : first evaluation after 4 years]. / Dépistage néonatal de la mucoviscidose en région liégeoise : première évaluation après 4 ans.

Since January 2020, neonatal screening for cystic fibrosis (CF-NBS) has been implemented in the Wallonia-Brussels Federation. It's based on the immunoreactive trypsin (IRT1) assay between day 2 and day 4, associated with a 12 CFTR pathogenic variants analysis and with an IRT control on day 21. The aim of this study is to evaluate the performance of our CF-NBS in Liège according to the quality criteria defined by the European Cystic Fibrosis Society's working group on neonatal screening. After four years, 58.762 newborns have been screened. Nineteen children with cystic fibrosis were diagnosed : 14 by NBS, 3 following a meconium ileus, 1 by family history and 1 false negative diagnosed on clinical basis. Furthermore, 39 healthy carriers and 2 uncertain diagnosis (CFSPID) were identified. The sensitivity of CF NBS is 93,3 % (target ≥ 95 %), the positive predictive value (PPV) 17,7 % (target ≥ 30 %). Increasing the TIR1 threshold by 0,1 in 0,1 from P99 to P99,5, would be associated with a lower sensitivity and a non-significant improvement of PPV. A national assessment of CF NBS needs to be carried out.

Depuis janvier 2020, un dépistage néonatal de la mucoviscidose a été officiellement implémenté en Fédération Wallonie-Bruxelles. Ce dépistage se base sur le dosage de la trypsine immunoréactive (TIR1) entre le 2ème et le 4ème jour de vie, associé à la recherche de 12 variants pathogènes du gène de la mucoviscidose (CFTR) et à un filet de sécurité consistant en un contrôle éventuel du dosage de TIR au jour 21. Le but de cette étude est d'évaluer la performance du dépistage en région liégeoise selon les critères de qualité définis par le groupe de travail sur le dépistage néonatal de la Société Européenne de la Mucoviscidose. Au terme de quatre années de dépistage officiel, 58.762 nouveau-nés ont été testés. Dix-neuf enfants atteints de mucoviscidose ont été diagnostiqués : 14 via le dépistage néonatal, 3 dans un contexte d'iléus méconial, 1 en raison d'une histoire familiale suggestive, 1 faux-négatif du dépistage diagnostiqué sur base clinique. De plus, 39 porteurs sains et 2 bébés avec un diagnostic incertain (CFSPID) ont été également dépistés. Le taux de sensibilité du dépistage est de 93,33 % (cible ≥ 95 %), la valeur prédictive positive (VPP) de 17,7 % (cible ≥ 30 %). L'élévation du seuil de TIR1 de 0,1 en 0,1, en allant du P99 au P99,5, est associée à une moins bonne sensibilité et à une amélioration non significative de VPP. Une évaluation du dépistage néonatal de la mucoviscidose à l'échelon national est à réaliser.

Analytical and clinical validation of a NGS panel in detecting targetable variants from ctDNA of metastatic NSCLC patients.

BACKGROUND: Circulating tumor DNA (ctDNA) has emerged as a promising biomarker for noninvasive cancer diagnostics, particularly in the context of metastatic non-small-cell lung cancer (NSCLC). Detecting targetable variants through ctDNA analysis offers the potential to guide treatment decisions, especially in cases where tissue samples are insufficient or unavailable. METHOD: In this study, we developed and validated a next-generation sequencing panel targeting 101 cancer-related genes (101-test) to detect somatic variants in ctDNA from a large cohort of Chinese patients with metastatic NSCLC. The performance of the 101-test was assessed by evaluating its limit of detection (LOD), accuracy, and precision in identifying molecular variants. Additionally, the concordance between ctDNA and tissue samples for detecting targetable variants was analyzed in 904 patients. RESULTS: The 101-test demonstrated a LOD of 0.38% for single-nucleotide variants (SNVs), 0.33% for insertions and deletions (InDels), and 0.33% for fusions. Sensitivity was 98.3% for SNVs, 100% for InDels, and 100% for fusions when compared to digital droplet PCR (ddPCR)/breakpoint PCR reference methods. The by-variant sensitivity for somatic variants was 97.5%, with a specificity of 99.9% between tumor-only and tumor-normal analyses. In a real-world cohort, the concordance between ctDNA and tissue samples for identifying targetable variants was 72.2% (457/633). Notably, the EGFR S768I variant, recently recommended by clinical guidelines, achieved an 80% concordance rate. Furthermore, 4.3% of patients (27/633) with targetable variants were identified exclusively through ctDNA testing. CONCLUSION: The ctDNA-based 101-test is a highly sensitive and specific tool for detecting targetable variants in metastatic NSCLC, particularly in cases with insufficient tissue samples. The findings support the use of ctDNA testing as a reliable and complementary method to traditional tissue-based molecular analysis, enhancing the precision of treatment strategies for NSCLC patients.

Evaluating diagnostic accuracy and agreement of TI-RADS scoring in thyroid nodules: A comparative analysis between sonographers and radiologists.

OBJECTIVE: The Thyroid Imaging Reporting and Data System (TI-RADS) is an essential tool for assessing thyroid nodules, primarily used by radiologists. This study aimed to compare the agreement of TI-RADS scores between sonographers and radiologists and to assess the diagnostic performance of these scores against histological findings in suspicious thyroid nodules. METHODS: In a retrospective analysis, 168 patients with suspicious thyroid nodules classified as TR3 and above by the radiologists were included. Both sonographers and radiologists independently assigned the American College of Radiologists (ACR) TI-RADS scores, which were then compared for inter-reader agreement using Cohen's Kappa statistic. The scores were also evaluated for diagnostic performance against histological results based on the Bethesda system. RESULTS: The study revealed a moderate overall agreement between sonographers and radiologists in TI-RADS scoring (κ = 0.504; 95% CI: 0.409-0.599), with poor agreement noted specifically for nodule margin scores (κ = 0.102; 95% CI: -1.430-0.301). In terms of diagnostic performance against histological outcomes, sonographers' TI-RADS scores showed a sensitivity of 100% and a specificity of 44.6%, while radiologists' scores showed a sensitivity of 100% but a lower specificity of 29.3%. CONCLUSION: The findings indicate moderate agreement in TI-RADS scoring between sonographers and radiologists, with reproducibility challenges especially in scoring nodule margins. The marginally superior diagnostic performance of sonographers' scores suggests potential efficiency benefits in involving sonographers in preliminary assessments. Future research should aim to encompass a wider range of TI-RADS categories and focus on minimizing scoring variability to enhance the system's clinical utility.

Optimizing antigen preparation for oxalyl-CoA decarboxylase enzyme diagnostic kit and ELISA system cutoff determination.

The prevalence of kidney stone disease is increasing globally, with calcium oxalate stones being the most common type. Oxalyl-CoA decarboxylase (OXC), an enzyme produced by the gut bacterium Oxalobacter formigenes, plays a crucial role in oxalate metabolism. Deficiencies in OXC activity can lead to the accumulation of oxalate, contributing to kidney stone formation. This study aimed to develop a reliable diagnostic assay for OXC by optimizing antigen production and establishing a cutoff value for an enzyme-linked immunosorbent assay (ELISA). We cloned, expressed, and purified recombinant OXC protein in Escherichia coli BL21(DE3), and generated specific polyclonal antibodies in rabbits. The ELISA system was optimized and validated using serum samples from 40 healthy individuals and 6 patients with oxalate-related disorders. The cutoff value was determined using the formula (M + 2SD), where (M) is the mean and (SD) is the standard deviation of the healthy sample results. The calculated cutoff value of 0.656750 effectively distinguished between healthy and affected individuals, with a sensitivity of 97.5% and a specificity of 83.3%. These findings provide a valuable tool for the early detection and management of oxalate-related disorders, with significant implications for clinical practice.

Investigation of serum Metrnl levels in pregnant women with gestational diabetes mellitus: a prospective non-interventional cohort study.

OBJECTIVE: The objective of this study was to investigate serum Metrnl levels in pregnant women with gestational diabetes mellitus and compare them with pregnant women without gestational diabetes mellitus. METHODS: The gestational diabetes mellitus group consisted of 87 pregnant women diagnosed with gestational diabetes mellitus, and the control group consisted of 93 healthy pregnant women without gestational diabetes mellitus. Serum Metrnl levels were determined by the enzyme-linked immunosorbent assay method. RESULTS: The two groups were similar in terms of demographic features. The median serum Metrnl level was found to be 1.16 ng/mL in the gestational diabetes mellitus group, while it was determined as 2.2 ng/mL in the control group (p=0.001). The two groups were divided into two subgroups based on participants' body mass index, normal weight and overweight. The lowest median Metrnl level was detected in the normal weight gestational diabetes mellitus group, followed by the overweight gestational diabetes mellitus group, normal weight control group, and overweight control group (1.1, 1.2, 2, and 2.4 ng/mL, respectively). Receiver operating curve analysis was performed to determine the value of the serum Metrnl level in terms of predicting gestational diabetes mellitus. The area under the curve analysis of serum Metrnl for gestational diabetes mellitus estimation was 0.768 (p=0.000, 95%CI 0.698-0.839). The optimal cutoff value for serum Metrnl level was determined as 1.53 ng/mL with 69% sensitivity and 70% specificity. CONCLUSION: Serum Metrnl levels in pregnant women with gestational diabetes mellitus were found to be significantly lower than in pregnant women without gestational diabetes mellitus. The mechanisms underlying the decrease in serum Metrnl levels in gestational diabetes mellitus remain unclear for now, and future studies will reveal the role of Metrnl in the pathophysiology of gestational diabetes mellitus.

Amplification-free detection of Ascochyta blight in chickpea using a simple molecular beacon assay.

Ascochyta blight is a major biotic stress that limits chickpea production globally. Fungicide application remains one of the effective control measures for the endemic spread. Due to the serious threat that synthetic fungicides pose to crop quality, early diagnosis of the pathogen is imperative. Whilst there have previously been several conventional lab-based diagnostic methods developed for early detection of Ascochyta rabiei, they require long assay times, specialised equipment and facilities, and trained personnel to process the samples. To overcome this challenge, a rapid amplification-free detection assay using a molecular beacon probe was developed. The method consists of a simple assembly assay that accurately detects pathogen within 30 min. The developed assay is species-specific and has a similar sensitivity level as conventional amplification-based methods. Although it is still a lab-based technique, considering the simplicity of the assay, it has a great potential to be developed further as a reliable in-field diagnostic device for early detection and quantification of fungal pathogen spores.

Highly sensitive and specific electrochemical biosensor for direct detection of hepatitis C virus RNA in clinical samples using DNA strand displacement.

Hepatitis C virus (HCV) is a common blood-borne infection that can lead to long-term illnesses such as hepatocellular cancer and liver cirrhosis. Early diagnosis is crucial for effective management, as no vaccine is available for preventing HCV infection. However, the high cost and complexity of current molecular diagnostic tools hinder efforts to achieve early diagnosis and prevent transmission, particularly in resource-limited settings. We developed a novel electrochemical biosensor for point-of-care testing (POCT) of HCV RNA. The sensor utilizes a strand displacement method, where the target RNA displaces a gold nanoparticle-labeled reporter probe (AuRP) from a pre-hybridized duplex with a magnetic nanoparticle (MNP)-labeled capture probe. The amount of displaced AuRP, detected using differential pulse anodic stripping voltammetry (DPASV), is directly proportional to the target RNA concentration. The biosensor exhibited excellent analytical performance, with a detection limit of 4 fM for synthetic targets and 43 ng/µL for RT-PCR products. Importantly, it successfully detected HCV RNA directly in clinical plasma samples without the need for RNA extraction or amplification. The sensor was used to analyze 30 RNA samples from HCV-positive patients, 20 cDNA samples from viral RNA, 30 HCV-positive plasma samples, and 22 HCV-negative plasma samples. The sensor results show good concordance with the RT-PCR results, demonstrating the sensor's potential for detecting HCV in clinical samples.