High Diagnostic Specificity Research Articles

A Study on the Role of LPA and ATX in Breast Cancer in Iraq

Breast cancer is the most common malignancy among women and a leading cause of cancer-related mortality. Early detection is crucial for improving treatment outcomes. This study investigates the roles of lysophosphatidic acid (LPA) and autotaxin (ATX) in the early diagnosis of breast cancer among Iraqi women. A case-control study was conducted involving 75 women diagnosed with breast cancer and 75 healthy controls. Blood samples were collected, and biochemical parameters, including LPA and ATX levels, were measured using the ELISA technique. Statistical analyses were performed with SPSS version 24, and receiver operating characteristic (ROC) analysis was used to evaluate diagnostic capabilities. The majority of breast cancer patients were aged 50-59 years (33.3%). Histologically, invasive ductal carcinoma was the most prevalent subtype (82.6%). Biochemical analysis revealed significant differences in alanine aminotransferase, total serum bilirubin, and alkaline phosphatase levels between patients and controls. LPA levels were significantly elevated in the patient group (868.48±142.11 pg/ml) compared to controls (212.01±54.94 pg/ml), while ATX levels were also higher (2252.20±399.46 pg/ml vs. 951.40±209.21 pg/ml). ROC analysis indicated that both LPA and ATX exhibited high diagnostic sensitivity (98%) and specificity (100%). In Iraqi women, elevated serum levels of LPA and ATX may serve as potential diagnostic biomarkers for breast cancer, highlighting their role in disease progression. Further studies are warranted to explore their clinical applications in breast cancer diagnosis and management

A label-free gold nanoparticles functionalized peptide dendrimer biosensor for visual detection of breakthrough infections in COVID-19 vaccinated patients

Given the global implementation of effective COVID-19 vaccines, which do not confer complete immunity, it is crucial to monitor the occurrence of breakthrough infections, particularly against newly emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants. Hence, we developed a label-free colorimetric assay using gold nanoparticles (GNPs) functionalized with a peptide dendrimer incorporating highly reactive epitopes of the nucleocapsid (N) protein. This assay relies on the tween-20 induced colorimetric changes caused by the aggregation of peptide dendrimer-coated GNPs in the absence of anti- SARS-CoV-2 N antibodies, and vice versa. Transmission electron microscopy, dynamic light scattering, and circular dichroism spectroscopy analyses all showed the formation of a uniform and highly stable coating of the peptide dendrimer over GNPs. Surface plasmon resonance experiments have demonstrated a strong binding affinity for the peptide dendrimer and anti- SARS-CoV-2 N antibodies, with a KD value of 525 nM. To validate the proof-of-concept, we have tested this assay on seventy human serum samples, and receiver operating characteristic curve analysis demonstrated high diagnostic sensitivity (88.89 %) and specificity (100 %). This approach opens up new avenues for the development of simple and rapid diagnostic assays for identifying antibodies against viral infections and other pathogens.

Evaluation of the AMP rapid test ROTA/ADENOVIRUS for simultaneous detection of rotavirus and adenovirus in stool samples.

Rotavirus (RV) and Adenovirus (AdV) gastroenteritis affect children worldwide. The sensitivity and specificity of the AMP Rapid Test ROTA/ADENOVIRUS [AMP-RA] for the detection of RV and AdV were compared against qPCR Allplex™ GI-Virus Assay [Allplex-GI]. A cross-sectional hospital-based surveillance study was conducted using stool samples from 1,148 patients under the age of five years between July 2023 and May 2024 with gastroenteritis. All samples were tested using the AMP-RA and challenged with the Allplex-GI that detects rotavirus A, enteric adenovirus F, norovirus genogroups GI & GII, sapovirus, and astrovirus. RV samples testing positive by AMP-RA and/or Allplex-GI were subjected to genotyping. Of the 1,148 stool samples, 123 samples tested positive for RV on AMP-RA while 133 tested positive on the Allplex™ GI. The clinical sensitivity and positive predictive value (PPV) of the AMP-RA for the detection of RV were 92.4% and 100%, respectively, while, the clinical specificity and negative predictive value (NPV) of the AMP-RA kit were 100% and 99%, respectively. The AMP-RA was able to detect all RV genotypes that circulated (G1[P8], G2[P4], G9[P8], G9[P4], G9[P9]). The AMP-RA assay detected 48 positive AdV samples and failed to detect 28 samples. Thus, the clinical sensitivity and PPV were 63.1% and 24.6%, respectively. 147 samples were positive for AdV on AMP-RA and negative by Allplex-GI, indicating a clinical specificity and NPV of 86.2% and 97.1%, respectively. AMP-RA showed high diagnostic sensitivity and specificity for RV detection. The reliability of detecting AdV was insufficient, emphasizing the need for further adenovirus test improvement.

Diagnostic Performance of Shear-Wave Elastography in Autoimmune Hepatitis: Evaluating Baseline Characteristics and Accuracy Across Fibrosis Stages.

Background Autoimmune hepatitis (AIH) is a chronic inflammatory condition affecting the liver, ultimately leading to fibrosis, followed by cirrhosis and liver failure. Although liver biopsy is necessary to confirm the disease and outline different stages of liver fibrosis, it is associated with several risks due to its invasiveness. Shear-wave elastography (SWE) has recently emerged as a non-invasive imaging modality, representing liver stiffness and providing additional fibrosis assessment. This study aims to validate SWE for diagnosing and discriminating liver fibrosis stage in AIH patients. Methodology This retrospective cohort study was conducted at the Sindh Institute of Urology and Transplantation, Karachi, with the enrollment of 162 patients diagnosed with AIH between March 2022 and December 2023. All participants underwent SWE and liver biopsy. The stages of fibrosis were assessed using the Metavir scoring system. Demographic, biochemical, and serological data were analyzed, including autoantibody and serum IgG levels. Diagnostic accuracy, sensitivity, and specificity of SWE were compared with histopathological findings. Results Of the 162 patients, 71 (43.8%) were males, and 91 (56.2%) were females, with a mean age of 35.8 ± 16.6 years. SWE had an overall diagnostic accuracy of 92.11%, sensitivity of 82.86%, and specificity of 98.7% in predicting fibrosis stages. SWE was effective in differentiating early fibrosis (F1-F2), while it showed reduced sensitivity for advanced fibrosis (F3-F4). Female gender (p = 0.009), elevated bilirubin (p = 0.009), autoantibody titers, and serum IgG levels (p = 0.043) were significant factors related to advanced fibrosis. Conclusions SWE has shown high diagnostic accuracy and specificity for the estimation of liver fibrosis in AIH, especially in the early stages. Sensitivity is lowered in advanced fibrosis. SWE is a very valuable non-invasive alternative to liver biopsy. Its use may decrease procedural risks, accelerate diagnosis, and improve outcomes in AIH. Further multicenter studies are recommended for confirmation and to explore the comparative performance of SWE against other non-invasive approaches.

Diagnostic Accuracy of Computational Fluid Dynamics-Based Fractional Flow Reserve Derived From Coronary Angiography: The ACCURATE Study.

Although fractional flow reserve (FFR) is the contemporary standard to detect hemodynamically significant coronary stenosis, it remains underused for the need of pressure wire and hyperemic stimulus. Coronary angiography-derived FFR could break through these barriers. The aim of this study was to assess the feasibility and performance of a novel diagnostic modality deriving FFR from invasive coronary angiography (AccuFFRangio) for coronary physiological assessment. The ACCURATE (Angiography-Derived Fractional Flow Reserve for Functional Evaluation of Coronary Artery Disease) study was a prospective, multicenter study conducted at 5 centers. Patients who had at least 1 lesion with a diameter stenosis of 30% to 90% were eligible. AccuFFRangio was measured on site in real time and compared with invasive FFR measurements in a blinded fashion. Primary end point was the diagnostic accuracy of AccuFFRangio in identifying functional relevant lesions. Between November 2020 and June 2021, pairwise analyses of AccuFFRangio and FFR were performed in 304 coronary arteries. AccuFFRangio showed good correlation (r=0.89; P<0.001) and agreement (mean difference: 0.01±0.06) with FFR. The diagnostic accuracy was 95.07% (95% CI, 91.99%-97.21%), which were significantly exceeded the prespecified target value (P<0.001). The sensitivity, specificity, and area under the receiver operating characteristic curve of 95.83% (95% CI, 89.67%-98.85%), 94.71% (95% CI, 90.73%-97.33%), and 0.972 (95% CI, 0.947-0.988), respectively. AccuFFRangio derived from coronary angiography alone has high diagnostic accuracy, sensitivity, and specificity compared with FFR. AccuFFRangio bears the potential for increasing the adoption of functional assessment of coronary artery stenosis and improving the use of physiological guided decision-making. URL: https://www.clinicaltrials.gov; Unique identifier: NCT04814550.

A triple protein-based ELISA for differential detection of ASFV antibodies.

African swine fever (ASF) caused by the ASF virus (ASFV) is a severe and highly contagious viral disease that poses a significant threat to the global pig industry. As no vaccines or effective drugs are available to aid prevention and control, early detection is crucial. The emergence of the low-virulence ASFV strain not expressing CD2v/MGFs (ASFVΔCD2v/ΔMGFs) has been identified domestically and internationally and has even become an epidemic in China, resulting in a complex epidemic. The commercialized ASFV ELISA kits available can detect the presence of ASFV infection in pigs, but they are unable to distinguish wild-type ASFV from gene-deleted strains. The current published ELISA assays can distinguish between the wild-type and CD2v gene-deleted ASFV but cannot differentiate wild-type and MGF505 gene-deleted ASFV or CD2v and MGF505 double-gene deleted ASFV infection, posing new challenges for an effective prevention and control of ASFV. In this study, the ASFV-p30, ASFV-CD2v, and ASFV-MGF505 proteins were expressed using a prokaryotic expression system, and a triple protein-based ELISA antibody detection method based on these proteins was successfully established to effectively differentiate between wild-type ASFV and ASFVΔCD2v and/or ASFVΔMGF505 virus infection. This triple protein-based ELISA showed good analytical specificity without cross-reactivity with antibodies against PRRSV, CSFV, PRV, and PCV2. Moreover, it demonstrates remarkable analytical sensitivity by allowing the identification of clinical samples even at dilutions as high as 1:800. The coefficient of variation the intra-assay and inter-assay were below 5%, indicating strong repeatability and reproducibility. To evaluate the performance of the triple protein-based ELISA, a total of 59 clinical serum samples were detected using the triple protein-based ELISA. The results showed that 22 samples were positive for ASFV, of which 19 were ASFV wild-type, one was ASFVΔCD2v, and two were ASFVΔMGF505. Compared with the commercialized triplex qPCR kit, the triple protein-based ELISA exhibited high diagnostic sensitivity and diagnostic specificity. The test accuracy with the commercialized triplex qPCR kit was 98.31% (58/59), and the test accuracy with the commercialized ELISA kit was 96.61% (57/59). These results indicated that the developed triple protein-based ELISA performs well in detection and differentiation. Therefore, it will be useful for the ASFV serological differential diagnosis and epidemiology study.

Diagnostic Accuracy of Active MMP-8 Point-of-Care Test in Peri-Implantitis.

This study aimed to evaluate the diagnostic sensitivity and specificity of the active matrix metalloproteinase-8 (aMMP-8) quantitative chairside point-of-care (PoC) lateral flow immunotest for peri-implant diseases, and it sought to correlate aMMP-8 levels with clinical parameters to determine its effectiveness as a biomarker for peri-implantitis. A cross-sectional study was conducted at the Department of Periodontology and Implant Biology, Aristotle University of Thessaloniki, Greece. Participants included systemically healthy individuals with at least one implant loaded for more than 1 year, who had not received periodontal treatment or antibiotics in the preceding 6 months. Exclusion criteria included diabetes and immune-compromising conditions. Peri-implant sulcular fluid (PISF) samples were collected from the mesiobuccal or distobuccal site of the implant. The quantitative chairside PoC lateral flow immunotest for peri-implant diseases (ImplantSafe test) and ORALyzer digital reader were used to analyze PISF, with results expressed in ng/mL. Clinical parameters such as bleeding on probing (BOP), probing depth (PD), recession (REC), and clinical attachment level (CAL) were measured at six sites per implant using a 15-mm scale periodontal probe. No significant differences were found in age, gender distribution, or smoking status between the healthy/mucositis and peri-implantitis groups. The peri-implantitis group showed significantly higher mean percentages of BOP (57.58 ± 31.73 vs. 18.79 ± 24.17), PD (4.59 ± 1.22 mm vs. 2.94 ± 0.78 mm), and CAL (5.21 ± 1.72 mm vs. 3.05 ± 0.81 mm). aMMP-8 levels were also significantly higher in the peri-implantitis group (53.39 ± 49.70 vs. 22.03 ± 32.87). The diagnostic test demonstrated a sensitivity of 81.25% and specificity of 74.07%, with an area under the curve of 79.6%, indicating overall good accuracy in distinguishing between positive and negative conditions. The aMMP-8 is a promising biomarker for peri-implantitis, showing elevated levels in affected patients. The aMMP-8 chairside test demonstrates high diagnostic sensitivity and specificity, supporting its use in early detection and monitoring of peri-implant diseases. Further research is needed to establish standardized protocols for its clinical application and to explore its long-term predictive value in implant care.

Role of the Lymphocyte Profile in Mediastinal Lymph Nodes in the Differential Diagnosis of Sarcoidosis and Tuberculous Lymphadenitis Patients Undergoing EBUS-TBNA.

The value of lymphocyte profiling (LP) in mediastinal lymph nodes for the differential diagnosis of sarcoidosis has not been extensively studied, and existing literature presents mixed results. This was a prospective study of patients with intrathoracic lymphadenopathy who underwent endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). LP in lymph node puncture fluid (LNPF) was evaluated using flow cytometry. The results of LP in sarcoidosis patients were compared with tuberculous lymphadenitis (TBLA) patients. Receiver operating characteristic (ROC) analysis was performed to determine the optimal cut-offs of the statistically significant parameters for screening for sarcoidosis. Based on the optimal cut-offs and the final diagnosis of sarcoidosis and TBLA, the sensitivity, specificity, and accuracy of every statistically significant parameter and different combinations of the above three parameters were calculated for the diagnosis of sarcoidosis. Forty-five cases of sarcoidosis and 33 cases of TBLA were enrolled in this study. Compared with the LP in TBLA patients, in sarcoidosis patients, the proportion of CD4 T cells and CD4/CD8 ratio increased, and the proportion of CD8 T cells and natural killer (NK) cells decreased. Among all single parameters, the CD4/CD8 ratio had high diagnostic sensitivity (84.4%), specificity (81.8%), and accuracy (83.3%) for sarcoidosis. Among all the combinations of three parameters, the combination of CD4, CD8, and NKT/NK ratio had high diagnostic sensitivity (91.1%), specificity (84.8%), and accuracy (87.2%) for sarcoidosis. Assessment of LP in LNPF may improve the differential diagnostic accuracy of sarcoidosis from TBLA and further strengthen the importance of LP in LNPF in the diagnostic workup of sarcoidosis.

Application of multiple inflammatory markers combined with PIVKA-II in differential diagnosis of AFP-NHCC

Alpha-fetoprotein (AFP) is a proven blood biomarker widely used in clinical detection of liver cancer, and its concentration increases immediately after liver injury, with high diagnostic specificity and low sensitivity. However, in patients with AFP-negative hepatocellular carcinoma (AFP-NHCC), also known as small liver cancer, their early stage of AFP expression is usually low or close to the normal range. Compared with AFP-positive HCC, AFP-NHCC is more likely to be subjected to missed diagnosis due to solely dependence on the measurement of blood AFP. Therefore, it is necessary to find a reliable, effective, and economical detection method for the early diagnosis of AFP-NHCC. PIVKA-II is closely related to the occurrence, development, invasion, and metastasis of liver cancer, and is also a new hematological marker widely used in the diagnosis of liver cancer in recent years. PIVKA-II effectively makes up for the limitation of negative AFP. 63.2% &amp;ndash; 76.3% of patients with negative AFP showed positive PIVKA-II, and if only PIVKA-II detection was relied on, there would still be missed diagnosis in early patients with AFP-NHCC. In this retrospective study, we selected several commonly used inflammatory indicators to explore the diagnostic efficacy of PIVKA-II, an abnormal form of prothrombin, combined with inflammatory indicators in patients with early-stage AFP-NHCC and analyzed the relationship between some clinical features and hematological indicators in patients with AFP-NHCC. Serum levels of high-sensitivity C-reactive protein (hs-CRP), prealbumin (PA), neutrophil/lymphocyte ratio (NLR), and PIVKA-II were compared among three groups (AFP-NHCC group, benign lesion group, and healthy subjects). The diagnostic efficacy of PIVKA-II alone for AFP-NHCC and the diagnostic efficacy of three inflammatory indicators combined with PIVKA-II for AFP-NHCC were calculated, and their diagnostic specificity and sensitivity were compared. Compared with the other two groups, the AFP-NHCC group showed significant changes in three inflammatory markers and PIVKA-II, which may be related to the inflammatory progression of the tumor. Therefore, we recommend the establishment of a laboratory-based detection approach for diagnosing early-stage AFP-NHCC by combining PIVKA-II with inflammatory indicators hs-CRP, PA, and NLR, to facilitate diagnosis, treatment, and prognosis of AFP-NHCC.

Deep learning-based automated measurement of hip key angles and auxiliary diagnosis of developmental dysplasia of the hip

ObjectivesAnteroposterior pelvic radiographs remains the most widely employed method for diagnosing developmental dysplasia of the hip. This study aims to evaluate the accuracy of an artificial intelligence model in measuring angles in pelvic radiographs of the hip. The assessment seeks to demonstrate the efficacy of the artificial intelligence model in diagnosing both developmental dysplasia of the hip and borderline developmental dysplasia of the hip through the analysis of pelvic radiographs.MethodsA total of 1,029 patients, including 273 men and 757 women, were retrospectively included in this study. The anteroposterior pelvic radiographs were randomly divided into three sets: the training set (720 cases), the validation set (103 cases), and the test set (206 cases). Key anatomical points on the anteroposterior pelvic radiographs were identified. The Sharp, Tönnis, and Center Edge angles were calculated automatically based on the corresponding criteria. The hip development status was compared between measurements obtained from the artificial intelligence model and those defined manually by two radiologists. The area under the receiver operating characteristic curve was utilized to assess the diagnostic performance of the artificial intelligence model.ResultsThe results obtained from both manual measurements and the artificial intelligence model demonstrated no significant differences in the Sharp, Tönnis, and Center edge angles (all p > 0.05). The intra-class correlation coefficients and correlation coefficient r values exceeded 0.75, indicating that both the artificial intelligence model and manual measurements exhibited good repeatability and a positive correlation. Notably, the artificial intelligence model provided measurements more faster than those conducted by radiologists (p = 0.001). The artificial intelligence model also demonstrated high diagnostic accuracy, sensitivity, and specificity for developmental dysplasia of the hip. The performance of the artificial intelligence model in diagnosing developmental dysplasia of the hip was robust. Additionally, the results from the artificial intelligence model and manual measurements were largely consistent with clinical diagnosis results (p = 0.01). The artificial intelligence model can effectively evaluate hip conditions by measuring the Sharp, Tönnis, and Center edge angles, which are consistent closely with clinical diagnosis results.ConclusionsThe results of the artificial intelligence model measurements demonstrate a high degree of consistency with those obtained through manual measurements. The angles of Sharp, Tönnis, and Center edge, as evaluated by the deep learning-based convolutional neural network model, exhibit robust diagnostic performance in identifying both developmental dysplasia of the hip and borderline developmental dysplasia of the hip. Consequently, the artificial intelligence model has the potential to fully replace manual measurements of these critical hip angles, providing a more efficient and precise alternative for diagnosing both conditions of the hip.

Анализ возможностей метода оценки кривой «поток–объем» по изменению ее формы при обструкции бронхов

In case of obstructive disorders, the flow–volume curve has a concave shape, but this feature is not given due attention. Тhe analysis of the velocity indicators of the respiratory function (such as the peak expiratory flow (PEF) and forced expiratory flows (FEFs)) will significantly expand the diagnostic capabilities of the spirometry method. This paper aims to perform a comparative analysis of the diagnostic strength of the methods of the flow-volume curve assessment by the changes in its shape in patients with obstructive airway diseases to determine the most reliable one. The respiratory function of 540 patients was tested (234 are men (57 [36; 67] years) and 306 are women (59 [44; 69] years)), with the ratio of areas under the actual curve and the predicted curve calculated for each one, as well as the angle formed by the curve; the ratio of the actual FEF (henceforth referred to as FEF) to the predicted FEF, cut-off points to differentiate between obstructive diseases and health. On the basis of these results, we concluded whether the patient’s bronchi were blocked. The results were then compared to the Knudson reference equations, with the test’s operational characteristics calculated compared to the standard. The methods of assessing the angle β and the total concavity of the flow-volume curve have high diagnostic sensitivity (87.8% and 95.6% respectively). The assessment of the area under the curve (AEX-FV) has high diagnostic specificity (88.6%). The results obtained show sufficient diagnostic efficiency of the methods of flow-volume curve estimation by the changes in its shape. However, the use of these methods in isolation from the reference equations does not currently seem reasonable for clinical practice. It appears reasonable to use the reference equations and one of the methods of curve shape assessment together.

Application of the Morphological Uterine Scoring Assessment in Ultrasound for Abnormal Uterine Bleeding.

Abnormal uterine bleeding (AUB) is a common gynecological condition that disrupts women's health due to irregularities in menstrual frequency, duration, and volume, often resulting in a significant impact on daily life and productivity. Accurate diagnosis of AUB is critical but complicated by its varied etiologies and presentations. Recent advancements in imaging techniques, particularly the Morphological Uterus Sonographic Assessment (MUSA), have enhanced the diagnostic precision of uterine pathologies such as fibroids and adenomyosis. MUSA combines gray-scale sonography, color Doppler, and three-dimensional ultrasound to evaluate uterine abnormalities with standardized terminology, ensuring diagnostic consistency. This study aimed to assess the efficacy of MUSA in diagnosing and managing AUB. A descriptive observational study was conducted on 50 patients at Dr. D. Y. Patil Medical College and Hospital, focusing on pre- and post-menopausal women with clinically symptomatic AUB. Patients underwent detailed ultrasonography, including both transabdominal and transvaginal scans, to evaluate uterine structures and correlate findings with histopathology. Results showed that 62% of patients had adenomyosis, while 38% had fibroids. MUSA effectively differentiated between the two conditions based on key ultrasound characteristics such as serosal contour, junctional zone, myometrial wall symmetry, and echogenicity. Adenomyosis cases showed significantly higher rates of heterogeneous echogenicity and asymmetrical myometrial walls compared to fibroids. Statistical analyses revealed high diagnostic sensitivity and specificity for both conditions, with an overall accuracy of 88.9% for adenomyosis and 94.1% for fibroids. The findings confirm the utility of MUSA in improving diagnostic accuracy and informing management strategies for AUB, particularly in complex cases. The study highlights MUSA as an indispensable tool for clinicians, facilitating enhanced patient outcomes through precise evaluation and treatment of uterine pathologies.

Detection of a Water-Soluble Hypericin Formulation in Glioblastoma Tissue with Fluorescence Lifetime and Intensity Using a Dual-Tap CMOS Camera System.

High hypericin-loaded polyvinylpyrrolidone (HHL-PVP) constitutes a novel approach to utilize the promising characteristics of hypericin for photodynamic diagnosis (PDD) and therapy (PDT) of brain tumors in an orally bioavailable formulation. The aim of this study was to investigate the ability of a Complementary Metal-Oxide-Semiconductor (CMOS) camera-based fluorescence imaging system to selectively visualize HHL-PVP in glioblastoma tissue even in the presence of 5-Aminolvevulinic acid (5-ALA) induced fluorescence, which is widely utilized in brain tumor surgery. We applied a previously established system with a non-hypericin specific filter for 5-ALA fluorescence visualization and a newly introduced hypericin-specific filter at 575-615 nm that transmits the spectrum of hypericin, but not 5-ALA fluorescence. Glioblastoma specimens obtained from 12 patients (11 with preoperative 5-ALA intake) were ex vivo incubated with HHL-PVP. Subsequently, fluorescence intensity and lifetime changes using both the non-hypericin specific filter and hypericin-specific filter were measured before and after HHL-PVP incubation and after subsequent rinsing. While no significant differences in fluorescence signal were observed using the non-hypericin specific filter, statistically significant increases in fluorescence intensity (p = 0.001) and lifetime (p = 0.028) after HHL-PVP incubation were demonstrated using the hypericin-specific filter. In consequence, specimens treated with HHL-PVP could be identified according to the fluorescence signal with high diagnostic sensitivity (87.5%) and specificity (100%). Our CMOS camera-based system with a hypericin-specific filter is capable of selectively visualizing hypericin fluorescence in glioblastoma tissue after ex vivo HHL-PVP incubation. In the future, this technique could facilitate clinical investigations of HHL-PVP for PDD and PDT while maintaining the current standard of care with 5-ALA guidance.

Microarray-based evaluation of selected recombinant timothy grass allergens expressed in E. Coli and N. Benthamiana

BackgroundTimothy grass (Phleum pratense) is a significant source of allergens, and recombinant allergens are increasingly used for diagnostic purposes. However, the performance of different recombinant allergen production systems in diagnostic assays needs further investigation to optimize their use in clinical settings.ObjectiveThe main objective of this study was to analyze and compare the diagnostic performance of recombinant timothy grass allergens produced in E. coli and N. benthamiana using a custom-made microarray chip.MethodsRecombinant timothy grass allergens Phl p 1, Phl p 2, Phl p 5, Phl p 6, Phl p 11, and Phl p 12 were produced in E. coli and/or N. benthamiana. A total of 113 patient serum samples were tested to evaluate the diagnostic sensitivity, specificity, inter-assay variability, and correlation of allergen-specific IgE detection compared to commercial multiplex tests (ALEX and ISAC). Additionally, the prevalence of sIgE to these allergens was assessed.ResultsPhl p 1, Phl p 2, Phl p 5, Phl p 6 and Phl p 11 showed high or very high positive correlation in immunoreactivity with other commercial multiplex tests. Notably, Phl p 11 fused with maltose-binding protein (MBP) demonstrated high diagnostic specificity and sensitivity, with a 0.3 arbitrary cut-off value. However, a high intra-assay variation was observed. The study also assessed specific IgE prevalence to timothy grass allergens within the tested patient cohort.ConclusionsRecombinant allergens from both E. coli and N. benthamiana demonstrated strong diagnostic potential on the microarray platform, with Phl p 11 (MBP-fused) showing particularly high performance. High intra-assay variation highlights the need for further optimization in allergen formulation and microarray storage conditions. These results highlight the potential of recombinant allergens for diagnostic applications, despite challenges with allergen stability in microarray formats. Specific IgE prevalence to timothy allergens revealed a sensitization profile consistent with findings from multiple studies.Graphical

A rapid economical multiplex PCR-RFLP method for molecular detection and genotyping of Giardia duodenalis clinical isolates

Giardia duodenalis is a common cause of diarrheal illness in regions with limited resources. The demand for rapid and cost-effective detection and genotyping methods in large-scale epidemiological studies and clinical diagnostics is imperative. Hence, we developed a multiplex PCR-RFLP technique targeting the tpi gene of G. duodenalis. The assay successfully screened G. duodenalis positive clinical samples (6.33 %; 36/565). It was also able to categorize the isolates into assemblages A (41.66 %; 13/36) and B (58.33 %; 23/36), as well as into subassemblages: AI (13.8 %; 5/36), AII (27.77 %; 10/36), BIII (36.11 %; 15/36) and BIV (22.22 %; 8/36). High diagnostic sensitivity (94.2 %), specificity (100 %) and accuracy (97.1 %) of the PCR assay were obtained, indicating its reliability for diagnosing giardiasis. Notably, the assay demonstrated close concordance with microscopy (κ=0.85) and reference PCR (κ=0.98) results. The optimized method offers a cost-effective and rapid approach for G. duodenalis detection and genotyping, convenient for epidemiological studies and clinical diagnostics.

Pulmonary embolism detection without intravenous contrast using electron density and Z-effective maps from dual-energy CT

Abstract Purpose This study aims to evaluate the feasibility of using electron density (ED) maps combined with Z-effective (Zeff) images obtained from unenhanced dual-layer dual-energy CT (dl-DECT) scans of the chest for the detection of pulmonary embolism (PE). Materials and methods A retrospective analysis was conducted on consecutive patients who underwent for contrast-enhanced chest CT (CECT) clinically suspected of PE or acute aortic syndrome. These scans were performed on a single dl-DECT scanner between October 2021 and November 2023. To distinguish emboli from circulating blood, color-coded maps were generated from the ED dataset superimposed on Zeff images, which were acquired from the unenhanced phase. Two radiologists with different levels of expertise independently assessed the presence of PE in the generated ED-Zeff maps, blinded to CECT results, which served as the reference standard. Diagnostic accuracy of ED-Zeff maps was assessed for each reader. Results The final study cohort comprised 150 patients, with 92 males (mean age: 68 ± 10 years, range: 47-93 years) and 58 females (mean age: 66 ± 15 years, range 38-89 years). ED-Zeff maps demonstrated high diagnostic performance, yielding accuracy, sensitivity, and specificity, respectively, of 86.67% (113/150, 95% CI, 80.16%-91.66%), 85% (17/20, 95% CI, 79.89%-92.19%), and 86.92% (113/130, 95% CI, 79.89%-92.19%). Ed-Zeff maps were able to identify PE in 85% of positive cases. Cohen’s kappa coefficient indicated excellent intra- and interobserver agreement (κ ≥ 0.9). Conclusion ED maps combined with Zeff images from unenhanced dl-DECT scans represent a feasible tool for detecting PE and may prove useful in evaluating patients with contraindications to iodinated contrast.

ThP8.7 - Giant Cell Arteries: Diagnostics

Abstract Aim Giant cell arteritis (GCA) poses a risk of severe ischaemic complications such as irreversible blindness which mandates early recognition of condition. We aimed to evaluate effectiveness of ultrasound doppler (USD) in diagnosing GCA compared to temporal artery biopsy (TAB) and American College of Rheumatology (ACR) score. Method This retrospective study included patients with suspected GCA who had consecutive TAB and USD examinations performed over a course of three years (2017-2022) which was correlated with ACR score. The data was also analysed to assess further management. Results There were 64 patients over this period who had both studies performed (22 were excluded as they did not have TAB) with 64% having an ACR score ≥3. Twenty males and forty-four females with a median age of 77 years (IQR 84-67). Overall, 81% patients had a negative USD result. Comparing USD results with TAB gold standard, there was sensitivity of 63% and specificity of 84%. Comparing USD results with ACR as gold standard, there was sensitivity of 20% and specificity of 83%. Forty percent patients with negative doppler result were continued on treatment mainly due to improvement of symptoms in response to steroids. Conclusion US Doppler has high diagnostic specificity and low sensitivity which makes it time-efficient, cost-effective and allows for the implementation of fast-track management pathway for majority patients. In certain circumstances, as cases of low clinical suspicion and positive doppler result, an additional test (TAB) would be required to confirm the diagnosis and cases where clinical suspicion is high with negative doppler, TAB should be considered.

Intestinal Anti-Endomysium Antibodies Are a Useful Tool for Diagnosing Celiac Disease in Pediatric and Adult Patients.

Intestinal anti-endomysium antibodies are a specific marker of celiac disease. The diagnostic accuracy of this marker seems high in pediatric patients and has not yet been investigated in adults, so the aim of this prospective multicentric study was to evaluate the specificity and sensitivity of this marker in childhood and adulthood. Pediatric and adult patients undergoing intestinal endoscopy for any intestinal condition were enrolled. Serological celiac disease markers and HLA type were evaluated in all patients. Intestinal biopsies were analyzed for standard histology and for intestinal anti-endomysium antibodies with biopsy culture assay. In this study, 291 patients (145 adults and 146 children) were included. In the adult population, 34 were diagnosed with celiac disease, 105 were controls, and, in 6, celiac disease was not confirmed. In the pediatric population, 77 were diagnosed with celiac disease, 57 were controls, and, in 12, celiac disease was not confirmed. High diagnostic sensitivity and specificity of intestinal anti-endomysium antibodies were confirmed in children and additionally proven in adults. To conclude, we can affirm that intestinal anti-endomysium antibodies can be detected with high diagnostic accuracy in both children and adults. The implementation of this marker in the diagnostic work-up would help clinicians to correctly identify celiac disease.

A rapid LAMP assay for the diagnosis of oak wilt with the naked eye

BackgroundOak wilt disease, caused by Bretziella fagacearum is a significant threat to oak (Quercus spp.) tree health in the United States and Eastern Canada. The disease may cause dramatic damage to natural and urban ecosystems without management. Early and accurate diagnosis followed by timely treatment increases the level of disease control success.ResultsA rapid assay based on loop mediated isothermal amplification (LAMP) was first developed with fluorescence detection of B. fagacearum after 30-minute reaction time. Six different primers were designed to specifically bind and amplify the pathogen’s DNA. To simplify the use of this assay in the field, gold nanoparticles (AuNPs) were designed to bind to the DNA amplicon obtained from the LAMP reaction. Upon inducing precipitation, the AuNP-amplicons settle as a red pellet visible to the naked eye, indicative of pathogen presence. Both infected and healthy red oak samples were tested using this visualization method. The assay was found to have high diagnostic sensitivity and specificity for the B. fagacearum isolate studied. Moreover, the developed assay was able to detect the pathogen in crude DNA extracts of diseased oak wood samples, which further reduced the time required to process samples.ConclusionsIn summary, the LAMP assay coupled with oligonucleotide-conjugated gold nanoparticle visualization is a promising method for accurate and rapid molecular-based diagnosis of B. fagacearum in field settings. The new method can be adapted to other forest and plant diseases by simply designing new primers.

'Positive' inter-ictal clinical signs of functional neurological disorders are found in patients with functional dissociative seizures.

Prior studies highlighted the high diagnostic specificity (ranging from 92% to 100%) of clinical signs observed in functional neurological disorders (FNDs). However, these signs are rarely looked for by epileptologists when trying to distinguish between functional dissociative seizure (FDS) and epileptic seizure. The aim of this study was to determine the prevalence of inter-ictal clinical signs of FND in a cohort of patients with probable FDS. The secondary objective was to compare the prevalence of inter-ictal FND clinical signs in FDS patients with age- and gender-matched epileptic patients without FDS. Patients diagnosed with FDS seen at two tertiary care centres and epileptic outpatients were included in the study. Each patient underwent a physical examination, searching for inter-ictal clinical signs of FND. In the FDS group, 79% of patients presented at least one sign of FND, compared to 16.6% of patients with epilepsy (p < 0.001). Moreover, 66.6% of FDS patients presented three or more FND signs, whereas only 4.1% of epileptic patients did (p < 0.001). The median number of FND clinical signs in the FDS group was four (SD 1.7; 5.5). Using the threshold of three signs or more, the specificity of detecting three or more FND signs was 83.3%, with a sensitivity of 79.2%. Inter-ictal clinical signs of FND are present in patients with FDS and should be looked for during neurological examination.