Routine Diagnostics Research Articles

Identification of Cuproptosis-Associated Prognostic Gene Expression Signatures from 20 Tumor Types.

We investigated the mRNA expression of 124 cuproptosis-associated genes in 7489 biopsies from 20 different tumor types of The Cancer Genome Atlas (TCGA). The KM plotter algorithm has been used to calculate Kaplan-Meier statistics and false discovery rate (FDR) corrections. Interaction networks have been generated using Ingenuity Pathway Analysis (IPA). High mRNA expression of 63 out of 124 genes significantly correlated with shorter survival times of cancer patients across all 20 tumor types. IPA analyses revealed that their gene products were interconnected in canonical pathways (e.g., cancer, cell death, cell cycle, cell signaling). Four tumor entities showed a higher accumulation of genes than the other cancer types, i.e., renal clear cell carcinoma (n = 21), renal papillary carcinoma (n = 13), kidney hepatocellular carcinoma (n = 13), and lung adenocarcinoma (n = 9). These gene clusters may serve as prognostic signatures for patient survival. These signatures were also of prognostic value for tumors with high mutational rates and neoantigen loads. Cuproptosis is of prognostic significance for the survival of cancer patients. The identification of specific gene signatures deserves further exploration for their clinical utility in routine diagnostics.

A-177 Using the Building Research Establishment Environmental Assessment Method to maximise the Environmental Social Governance benefits of building a pathology hub laboratory

Abstract Background Organisations focusing on Environmental Social Governance (ESG) factors do not just benefit society and the planet, they also enjoy better performance across key profitability metrics, driven by higher growth and productivity, lower operational costs and enhanced return on investment. SYNLAB’s new Synnovis Pathology Hub, built in partnership with the National Health Service (NHS), is a high-tech purpose-built laboratory building with construction completing in Spring 2024. It will serve more than 1.7 million people in the southeast London region delivering both routine and special diagnostics in support of a 15-year NHS pathology contract. The redevelopment of the site presented an opportunity to closely align SYNLAB company strategy with the delivery of its ESG ambitions, as well as achieve financial and operational sustainability. Methods BREEAM (Building Research Establishment Environmental Assessment Method) is a standardised framework within which SYNLAB could collaborate with the building landlord and Hub delivery partners. BREEAM assessment categories are: energy, land use and ecology, water, health and wellbeing, pollution, transport, materials, waste and management. Applying the intent of SYNLAB’s ESG strategy to the hub build programme, setting this expectation with stakeholders, and using BREEAM as a platform on which to track and report ESG benefits ensured ESG and related deliverables were high on the agenda to be captured and delivered. Results The hub is on track to achieve the BREEAM Excellent rating which means it will rank within the top 10% of commercial buildings in the UK in terms of ESG benefits. Contributors to this achievement include: Recycling over 85% of materials during construction, saving more than 500 tonnes of materials from going to waste; Incorporation of changing rooms and 56 cycle rack spaces to encourage healthy and sustainable travel, helping to keep London’s air clean and reduce CO2e emissions; An Energy Management System within the Building Management System providing detailed area by area analysis and control. Along with water metering per floor, this helps SYNLAB to understand usage and focus process improvements; An energy efficient air handling system, a key area of opportunity for laboratory energy efficiency; Conformity with EU Taxonomy for Sustainable Activities, for example by limiting the flow rate of water to prevent wastage; Reuse of materials within the building, for example by relocating seating, private call areas, glass walls; Reuse of materials externally through donations to charitable organisations, for example televisions and fridges were given to a local care home, improving the residents’ quality of life; Working with local schools and wider the community. Conclusions Incorporating BREEAM ESG criteria from the early stages of planning in construction projects provide a higher likelihood of stakeholder buy-in, adoption and delivery. Pathology networks are considered key in delivering the national ambitions set out within the Long-Term Plan for the NHS and there is increased government and social pressure driving healthcare businesses to adopt ESG. There is the opportunity within Pathology networking and Pathology new builds to harness these benefits from the outset and lay the long term foundations for ESG within healthcare for future sustainability.

B-077 F-actin as a New Marker Antigen on an Established Multiparametric Immunoassay Profile for Liver Diseases

Abstract Background Autoimmune hepatitis (AIH) and primary biliary cholangitis (PBC) are severe diseases with heterogenous manifestations, which affect people of all populations and age groups. Diagnosis and differentiation from other liver diseases such as primary sclerosing cholangitis (PSC) remain a substantial challenge for hepatologists. Since AIH and PBC are caused by different sets of autoantibodies, we previously developed a line blot for the parallel detection of all relevant parameters. We now added the AIH-specific antigen filamentous actin (F-actin) to this established liver immunoassay profile. Methods Sera from 15 patients with AIH, 15 patients with overlap AIH/PBC or AIH/PSC, 32 patients with PSC, 53 patients with PBC, 4 patients with non-alcoholic fatty liver disease/non-alcoholic steatohepatitis (NAFLD/NASH), 30 patients with hepatitis B infection and 79 patients with hepatitis C infection as well as sera from 150 blood donors were examined using the EUROLINE line immunoassay system (EUROIMMUN autoimmune liver diseases 9 Ag plus F-actin (IgG); contains the PBC-relevant antigens AMA-M2, M2-3E (BPO), Sp100, PML and gp210, the AIH-associated antigens LKM-1, LC-1 and SLA/LP and F-actin as well as Ro52). The newly included F-actin antigen was polymerized from human globular actin (G-actin) and assessed by sedimentation analysis and dynamic light scattering. The scan software EUROLineScan (EUROIMMUN) was used to automatically determine presence and intensity of the resulting bands on the blot. Results Quality assessment revealed successful polymerization of F-actin. Autoantibodies against this newly developed F-actin antigen were detected in 53.3% of sera from AIH patients and 40% of AIH+PBC/AIH+PSC overlap patients. 93% of these positive sera did not contain other autoantibodies associated with AIH. Only one PSC patient (3.1%), two patients with hepatitis C (2.5%) and two blood donors (1.3%) also showed anti-F-actin positivity. All tested sera of PBC, NAFLD/NASH and hepatitis B patients were evaluated as anti-F-actin negative. Conclusions The addition of F-actin to the established multiparametric EUROLINE profile for the simultaneous and monospecific detection of AIH- and PBC-relevant autoantibodies is a great advance for routine diagnostics of patients with symptoms associated with various liver diseases.

Coexistence of seven different carbapenemase producers in a single hospital admission screening confirmed by whole-genome sequencing

ObjectiveTo molecularly characterize several extensively drug-resistant isolates from a single hospital admission screening of a war injured patient from the Ukraine. MethodsAdmission screening included swabs from skin, wounds, catheters, nasopharyngeum and rectum. Bacterial identification, antimicrobial susceptibility testing (AST) and rapid multiplex PCR assays targeting resistance genes were performed during routine diagnostics. Isolates positive by PCR had their genomes sequenced using short- and long read-platforms (MiSeq and MinION) to confirm species, identify resistance genes and plasmids and investigate clonality with core genome MLST (cgMLST). ResultsSeven Gram-negative pathogens (Acinetobacter baumannii (n=2; ST78, ST2), Klebsiella pneumoniae (n=2; ST395), Pseudomonas aeruginosa (n=1; ST1047), Escherichia coli (n=1; ST46), Enterobacter cloacae complex (n=1; ST231)) were molecularly confirmed non-identical. AST showed resistance to carbapenems (7/7 isolates) and last resort treatment options such as ceftazidime-avibactam (6/7 isolates) and cefiderocol (4/7 isolates). All isolates were colistin susceptible. Sequencing identified the E. cloacae complex as E. hormaechei subsp. xiangfangensis. Six acquired carbapenemase genes (blaIMP-1, blaNDM-1, blaOXA-48, blaNDM-5, blaOXA-23 and blaOXA-72) were detected. Both A. baumannii isolates differed in sequence type, carbapenemases and cefiderocol susceptibility. Both K. pneumoniae isolates shared sequence type and some resistance genes on an IncR plasmid but were different in cgMLST and carbapenemases (OXA-48 or NDM-1). One vancomycin-resistant Enterococcus faecium was also detected (VanA). ConclusionWar injured patients from Ukraine may carry different clones of multidrug-resistant pathogens with limited treatment options and diverse resistance genes at risk for dissemination. Infection control measures should include early molecular characterization of isolates for detection of routes of transmission.

Stool Xpert® MTB/RIF Ultra for TB diagnosis in children: experience from a national scale-up programme.

We describe the programmatic scale-up of stool testing using Xpert® MTB/RIF Ultra (Ultra), as recommended by the WHO to improve childhood pulmonary TB (PTB) diagnosis. USAID's Alliance for Combating TB in Bangladesh Activity, in collaboration with the National TB Control Programme, is implementing a stool-based diagnostic approach at 51 healthcare facilities in Bangladesh to improve PTB detection. Specimens from children (<15 years) with presumptive TB were tested using 'stool Ultra' with routine TB diagnostics. Physicians confirmed TB diagnosis and provided treatment as per national guidelines. Between March 2022 and December 2023, 16,429 specimens were tested, 871 (5.3%) were positive, and 642 (73.7%) showed 'trace detected' results. Positivity was significantly higher among females, and children presented with 'only cough ≥2 weeks', 'cough ≥2 weeks + weight loss', or 'fever ≥2 weeks, weight loss, fatigue + contact history'. Positivity was higher among '10-14 years old' children; however, 'trace detected' was highest among '5-9 years', followed by children aged '>2-<5 years' and '0-2 years'. Testing stools using Ultra provides a more effective way of diagnosing bacteriologically positive PTB in children. However, positivity varies with presenting symptoms/criteria, highlighting the need for careful diagnostic evaluation to ensure optimum use of limited diagnostic resources.

Antimicrobial susceptibility testing of bone and joint pathogens using isothermal microcalorimetry.

The rise in osteomyelitis and periprosthetic joint infections, in combination with increasing life expectancy and the prevalence of diabetes, underscores the urgent need for rapid and accurate diagnostic tools. Conventional culture-based methods are often time-consuming and prone to false-negatives, leading to prolonged and inappropriate antibiotic treatments. This study aims to improve osteomyelitis diagnostics by decreasing the time to detection and the time to an antibiotic susceptibility result to enable a targeted treatment using isothermal microcalorimetry (IMC). IMC measures heat flow in real-time, providing insights into bacterial metabolism without the need for labeling. Using clinical isolates from bone infections, assessing their response to antibiotics through IMC, we demonstrated that IMC could detect bacteria within 4 h and determine antimicrobial susceptibility profiles within 2-22 h (median 4.85, range 1.28-21.78). This is significantly faster than traditional methods. A decision tree, based on antibiotic susceptibility, accurately categorized pathogens, achieving high accuracy (74-100%), sensitivity (100%), and specificity (65-100%). These findings suggest that IMC could redefine diagnostics of bone and joint infections and potentially infections in general, offering timely and precise treatment guidance, thereby improving patient outcomes and reducing health care burdens. Further optimization and clinical validation are needed to fully integrate IMC into routine diagnostics.

Immunopathology of Corneal Allograft Rejection and Donor-Specific Antibodies (DSAs) as Immunological Predictors of Corneal Transplant Failure.

Despite tremendous developments in the field of laboratory testing in transplantation, the rules of eligibility for corneal transplantation still do not include typing of human leukocyte antigens (HLAs) in the donor and recipient or detection of donor-specific antibodies (DSAs) in the patient. The standard use of diagnostic algorithms is due to the cornea belonging to immunologically privileged tissues, which usually determines the success of transplantation of this tissue. A medical problem is posed by patients at high risk of transplant rejection, in whom the immune privilege of the eye is abolished and the risk of transplant failure increases. Critical to the success of transplantation in patients at high risk of corneal rejection may be the selection of an HLA-matched donor and recipient, and the detection of existing and/or de novo emerging DSAs in the patient. Incorporating the assessment of these parameters into routine diagnostics may contribute to establishing immune risk stratification for transplant rejection and effective personalized therapy for patients.

DdPCR for the Detection and Absolute Quantification of Oropouche Virus.

Oropouche virus (OROV) is a segmented RNA virus belonging to the genus Orthobunyavirus in the family Peribunyaviridae. Herein, an in-house droplet digital PCR (ddPCR) assay was used for the detection and quantification of OROV. The ddPCR reaction was assessed as duplex assay using the human housekeeping gene RPP30. Limit of detection (LoD) analysis was performed in whole blood, serum, and urine. The assay was executed on a total of 28 clinical samples (whole blood n = 9, serum n = 11, and urine n = 8), of which 16 specimens were tested positive at the routine molecular diagnostics (endpoint and real-time PCRs). The LoD of the ddPCR performed using 10-fold serial dilution of OROV detected up to 1 cp/µL in all the biological matrices. Compared to the routine molecular diagnostics, the ddPCR assay showed 100% sensitivity for whole blood and serum and 75% for urine, highlighting higher positive rate of ddPCR. We have established a quantitative RNA detection method of OROV with high sensitivity and specificity based on ddPCR. This test is capable of quantitatively monitoring the viral load of OROV and can contribute, in addition to laboratory diagnosis, to shed light on the pathogenesis, filling in the knowledge gaps of this neglected disease and to the vector control programs.

High Prevalence of Chromosomal Rearrangements and LINE Retrotranspositions Detected in Formalin-Fixed, Paraffin-Embedded Colorectal Cancer Tissue

Structural variants (SVs) caused by chromosomal rearrangements in common fragile sites (CFS) or LINE retrotranspositions are highly prevalent in colorectal cancer (CRC). However, methodology for targeted detection of these SVs is lacking. We here report the use of formalin-fixed paraffin-embedded targeted locus capture (FFPE-TLC) sequencing as a novel technology for targeted detection of tumor-specific SVs. Analysis of 29 FFPE colorectal tumor and 8 matched normal samples revealed tumor-specific SVs in 24 patients (83%) with a median of 2 SVs per patient (range 1-21). 104 SVs were found in the CFS-associated genes MACROD2, PRKN, FHIT and WWOX in 18 patients (62%) and 39 SVs caused by three LINE transposable elements were found in 15 patients (52%). Tumor-specificity of SVs was independently verified by droplet digital PCR of tumor tissue DNA and their applicability as plasma circulating tumor DNA biomarkers was demonstrated. We conclude that FFPE-TLC sequencing enables the detection of tumor-specific SVs caused by chromosomal rearrangements and LINE retrotranspositions in FFPE tissue. Therefore, FFPE-TLC sequencing facilitates the investigation of the biological and clinical impact of SVs using FFPE material from (retrospective) cohorts of cancer patients and has potential clinical applicability to detect SV biomarkers in the routine molecular diagnostics setting.

The Role of Voluntary Counselling and Testing Points (VCTs) in HIV Diagnostics – Analysis of VCTs Activity in Poland in the Context of Local and European Epidemiology

Abstract Screening tests are now readily accessible, quick and highly sensitive. Diagnostics of HIV infections involves serological testing followed by confirmation tests by molecular biology methods (NAAT). Fast, accurate, anonymous and free HIV testing has been provided by Voluntary Counselling and Testing Points (VCTs) for over 20 years to any adult in Poland, regardless of nationality. Additionally, the educational activities are conducted aimed at informing people from various social backgrounds about HIV prevention and diagnostic possibilities, as well as other sexually transmitted infections (STIs). The aim of the article was to present the procedures for the operation of VCTs in Poland, their location, testing scheme and a summary of the activities that promote such points in the medical community. Between 2019 and 2023, 157,833 people registered for HIV testing in VCTs in Poland, of which 10,177 (6.45%) were foreigners. In the analyzed period, the number of tests among foreigners visiting VCTs in Poland almost doubled, from 1,552 in 2019 to 2,827 in 2023. In addition, a higher percentage of positive results was recorded among foreigners – 5.08% compared to Poles – 1.24%. The frequency analysis of positive results detection reported in VCTs vs. National Institute of Public Health NIH – National Research Institute showed that, on average 26% (2,342/8,891) of positive results in Poland from 2019–2023 were detected in the framework of tests carried out in VCTs. The operation of VCT points in Poland enables the reaching of a larger group of people, especially key adult populations. It is a valuable complement to routine diagnostics outside the health care system.

Corynebacterium pseudotuberculosis: Whole genome sequencing reveals unforeseen and relevant genetic diversity in this pathogen.

Corynebacterium pseudotuberculosis (CPS) is an important bacterial animal pathogen. CPS causes chronic, debilitating and currently incurable infectious diseases affecting a wide range of livestock and wild herbivores including camelids worldwide. Belonging to the Corynebacterium diphtheriae complex, this pathogen can also infect humans. The classical characterization of CPS is typically based on the testing of nitrate reductase activity, separating the two biovars Equi and Ovis. However, more refined resolutions are required to unravel routes of infection. This was realized in our study by generating and analyzing whole genome sequencing (WGS) data. Using newly created core genome multilocus sequence typing (cgMLST) profiles we were the first to discover isolates grouping in a cluster adjacent to clusters formed by CPS biovar Equi isolates. This novel cluster includes CPS isolates from alpacas, llamas, camels and dromedaries, which are characterized by a lack of nitrate reductase activity as encountered in biovar Ovis. This is of special interest for molecular epidemiology. Nevertheless, these isolates bear the genes of the nitrate locus, which are characteristic of biovar Equi isolates. However, sequence analysis of the genes narG and narH of the nitrate locus revealed indels leading to frameshifts and inactivity of the enzymes involved in nitrate reduction. Interestingly, one CPS isolate originating from another lama with an insertion in the MFS transporter (narT) is adjacent to a cluster formed by ovine CPS isolates biovar Equi. Based on this knowledge, the combination of biochemical and PCR based molecular biological nitrate reductase detection can be used for a fast and uncomplicated classification of isolates in routine diagnostics in order to check the origin of camelid CPS isolates. Further analysis revealed that partial sequencing of the ABC transporter substrate binding protein (CP258_RS07935) is a powerful tool to assign the biovars and the novel genomovar.

Identification and phylogenetic analysis of bovine herpesvirus 4 isolated from aborted bovine foetuses in Türkiye.

Bovine herpesvirus 4 (BoHV-4) is responsible for reproductive disorders and abortions in cattle. The aim of this study was to investigate the potential role of BoHV-4 in abortions in dairy cows when the possible aetiological cause could not be determined in Türkiye. The study also provided the molecular characterization of BoHV-4 strains based on the gB gene. In this study, foetuses (n = 188) and serum (n = 50) were retrospectively used for PCR and indirect ELISA assays, respectively. In addition, foetuses with BoHV-4 were tested for the isolation of the BoHV-4 strain in MDBK cell culture. The BoHV-4 gB gene was detected in the brain, liver, lung, kidney, and spleen tissues of three foetuses out of 188 aborted foetuses. The BoHV-4 strain was isolated from the kidney of one foetus in MDBK cell culture. The three aborted foetuses were from different cows in the same herd in 2021. On this dairy farm, the BoHV-4 seroprevalence rate in cattle with a history of abortion was 96% (48/50). Phylogenetic analysis revealed that the detected strains belong to genotype II. The present study shows that the impact of BoHV-4 on abortions in cattle should not be ignored. It can therefore be informative to focus on BoHV-4 in routine laboratory diagnostics for abortions without a proven aetiology or in cattle herds that have previously been infected with BoHV-4.

Comparison of clinical selection-based genetic testing with phenotype-agnostic extensive germline sequencing to diagnose genetic predisposition in children with cancer: a prospective diagnostic study

Germline data have become widely available in paediatric oncology since the introduction of paired tumour-germline sequencing. To guide best practice in cancer predisposition syndrome (CPS) diagnostics, we aimed to assess the diagnostic yield of extensive germline analysis compared with clinical selection-based genetic testing among all children with cancer. In this prospective diagnostic study, all children (aged 0-19 years) with newly diagnosed neoplasms treated in the Netherlands national centre, the Princess Máxima Center for Pediatric Oncology (Utrecht, Netherlands), between June 1, 2020, and July 31, 2022, were offered two approaches to identify CPSs. In a phenotype-driven approach, paediatric oncologists used the McGill Interactive Pediatric OncoGenetic Guidelines tool to select children for referral to a clinical geneticist, and for genetic testing. In a phenotype-agnostic approach, CPS gene panel sequencing (143 genes) was offered to all children. In children declining the research CPS gene panel, 49 CPS genes were still analysed as part of routine diagnostics by the pathologist. Children with a causative CPS identified before neoplasm diagnosis were excluded. The primary objective was to compare the number and type of patients diagnosed with a CPS between the two approaches. 1052 children were eligible for this study, of whom 733 (70%) completed both the phenotype-driven approach and received phenotype-agnostic CPS gene panel sequencing (143 genes n=600; 49 genes n=133). In 53 children, a CPS was identified: 14 (26%) were diagnosed by the phenotype-driven approach only, 22 (42%) by CPS gene sequencing only, and 17 (32%) by both approaches. In 27 (51%) of the 53 children, the identified CPS was considered causative for the child's neoplasm. Only one (4%) of the 27 causative CPSs was missed by the phenotype-driven approach and was identified solely by phenotype-agnostic CPS gene sequencing. In 26 (49%) children, a CPS with uncertain causality was identified, including 14 adult-onset CPSs. The CPSs with uncertain causality were mainly detected by the phenotype-agnostic approach (21 [81%] of 26). Phenotype-driven genetic testing and phenotype-agnostic CPS gene panel sequencing were complementary. The phenotype-driven approach identified the most causative CPSs. CPS gene panel sequencing identified additional CPSs, many of those with uncertain causality, but some with clinical utility. We advise clinical evaluation for CPSs in all children with neoplasms. Phenotype-agnostic testing of all CPS genes is preferably conducted only in research settings and should be paired with counseling. Stichting Kinderen Kankervrij.

Molecular Investigation of Small Ruminant Abortions Using a 10-Plex HRM-qPCR Technique: A Novel Approach in Routine Diagnostics.

The objective of this study was to apply and preliminarily evaluate a High-Resolution Melting (HRM) analysis technique coupled with qPCR, that allows the simultaneous detection of 10 different ruminant abortogenic pathogens, for investigating abortions in sheep and goats throughout Greece. A total of 264 ovine and caprine vaginal swabs were obtained the week following the abortion from aborted females and analyzed using a commercially available kit (ID Gene™ Ruminant Abortion Multiplex HRM, Innovative Diagnostics). Results indicated a high prevalence of Coxiella burnetii and Chlamydophila spp., which were detected in 48.9% and 42.4% of the vaginal swabs, respectively. Results for these most commonly detected pathogens were compared with those of a well-established commercial qPCR kit, with near-perfect agreement. Toxoplasma gondii, Salmonella spp., Brucella spp., Anaplasma phagocytophilum, Campylobacter fetus, and Neospora caninum were also identified, the two latter reported for the first time in the country in small ruminants. Mixed infections occurred in 35.6% of the animals examined. This technique allows for the simultaneous detection of many abortogenic pathogens in an accurate and cost-effective assay. Detection of uncommon or not previously reported pathogens in various cases indicates that their role in ovine and caprine abortions may be underestimated.

Genomic insights into prenatal diagnosis of congenital heart defects: value of CNV-seq and WES in clinical practice.

This study aimed to assess the efficiency of CNV-seq and WES in detecting genetic cause of congenital heart disease (CHDs) in prenatal diagnoses and to compare CNV detection rate between isolated and non-isolated CHD cases. We conducted a retrospective study of 118 Chinese fetuses diagnosed with CHD by prenatal ultrasound. Participants underwent CNV-seq and, if necessary, WES to detect chromosomal and single nucleotide variations. The overall detection rate for pathogenic or likely pathogenic chromosomal abnormalities was 16.9%, including 7.6% aneuploidies and 9.3% pathogenic/likely pathogenic copy number variations (CNVs), predominantly 22q11.2 deletion syndrome (54.4%). The sensitivity and specificity of CNV-Seq for detecting P/Lp CNVs were 95% and 100%, respectively. CNV-Seq offered a 6.7% improvement in detecting chromosomal abnormalities over karyotyping. WES further identified significant single nucleotide and small indel variations contributing to CHD in genes such as TMEM67, PLD1, ANKRD11, and PNKP, enhancing diagnostic yield by 14.8% in cases negative for CNVs. Non-isolated CHD cases exhibited higher rates of detectable chromosomal abnormalities compared to isolated cases (32.4% vs. 9.9%, p = 0.005), underlining the genetic complexity of these conditions. The combined use of CNV-seq and WES provides a comprehensive approach to prenatal genetic testing for CHDs, unveiling significant genetic cause that could impact clinical management and parental decision-making. This study supports the integration of these advanced genomic technologies in routine prenatal diagnostics to increase detection diagnostic yields of causal genetic variants associated with CHDs.

Demographic and socioeconomic obstacles to access to malaria services for Myanmar migrants in Thailand

BackgroundTypically mobile and vulnerable, migrants face significant barriers to access to routine malaria prevention, diagnostics and treatment, which leads to unchecked malaria transmission, particularly in border regions with a high population displacement. This study aimed to investigate the demographic and socioeconomic obstacles to access to malaria services among Myanmar migrants residing in the Thailand–Myanmar border areas.MethodsA cross-sectional study was conducted in early 2024 across three districts near the Thailand–Myanmar border. Quantitative data were collected from Myanmar migrants using standardized questionnaires through structured surveys. Data analysis included descriptive statistics and simple and multiple logistic regression models.ResultsOut of 300 participants, approximately a quarter (27.3%) reported adequate access to comprehensive malaria services, including prevention, diagnostics, treatment and malaria-related health information. In multiple logistic regression models, factors associated with inadequate access included Myanmar migrants aged over 60 years (aOR: 7.63, 95% CI 1.74–20.58), accompanied by one to three family members (aOR: 3.33, 95% CI 1.06–8.45), earning monthly incomes below 3000 THB (aOR: 5.13, 95% CI 1.38–19.09) and 3000 to 6000 THB (aOR: 3.64, 95% CI 1.06–12.51), belonging to the Karen ethnicity (aOR: 2.13, 95% CI 1.02–3.84), with poor perception toward malaria (aOR: 2.03, 95% CI 1.03–4.01) and with poor preventive and health-seeking practices (aOR: 5.83, 95% CI 2.71–9.55).ConclusionsA significant proportion of Myanmar migrants encounter demographic and socioeconomic barriers to access to routine malaria services in Thailand. Tailored interventions are required to expand such access, including the recruitment of worksite health volunteers, strengthening the role of ethnic health organizations across the border and collaboration with private sector stakeholders (e.g. farm/company owners) to distribute preventive tools and ensure timely referral of suspected malaria cases to health facilities.

Hypomania-Checklist-33: risk stratification and factor structure in a mixed psychiatric adolescent sample

BackgroundThe 33-item Hypomania Checklist (HCL-33) has been shown to distinguish between adolescent bipolar disorder (BD) and unipolar depression. To investigate the utility of the HCL-33 as a screening tool in routine diagnostics, the frequency and psychopathological characteristics of detected individuals in a mixed psychiatric sample necessitate more examination.MethodsThe HCL-33, Children’s Depression Inventory, Beck’s Anxiety Inventory, and Strengths and Difficulties Questionnaire were completed by 285 children and adolescents (12–18 years) in a mixed psychiatric sample. Applying the proposed HCL-33 cut-off score of ≥ 18, individuals with depressive symptoms were divided into at-risk or not at-risk for BD groups. The factorial structure, sum and factor score correlations with psychopathology, and impact on daily functioning were assessed.Results20.6% of the sample met at-risk criteria for BD. These individuals (n = 55) were older, more anxious, and showed more conduct problems vs the not at-risk group (n = 107). A two- and a three-factor model were pursued with the same Factor 1 (“active-elated”). Factor 2 (“risk-taking/irritable”) was separated into 2a (“irritable-erratic”) and 2b (“outgoing-disinhibited”) in the three-factor model. Whereas higher Factor 2 and 2a scores correlated with a broad range of more severe symptomatology (i.e., depression, anxiety, hyperactivity), higher Factor 1 and 2b scores correlated with more emotional and conduct problems, respectively. 51.7% of the sample reported a negative impact from hypomanic symptoms on daily functioning.LimitationsCross-sectional design and data collection in a single mental health service.ConclusionsThe HCL-33 may be a useful tool to improve diagnostics, especially in adolescents with depressive symptoms additionally presenting with anxious symptoms and conduct problems.

Modification of carbapenemase inhibition test and comparison of its performance with NG-Test CARBA 5 for detection of carbapenemase-producing enterobacterales.

Adequately and accurately identifying carbapenemase-producing Enterobacterales (CPE) is vital for selecting appropriate antimicrobial therapy and implementing effective infection control measures. This study aims to optimize the phenotypic detection method of carbapenemase for routine diagnostics in clinical microbiology laboratories. Carbapenemase genes in 2665 non-duplicate carbapenem-resistant Enterobacterales (CRE) clinical strains collected from various regions of China were confirmed through whole-genome sequencing (WGS). The carbapenemase inhibition test (CIT) was conducted and interpreted using different methods and breakpoints, then compared with the NG-Test CARBA 5 for carbapenemase detection. The diagnostic performance of the CIT method was optimal when the carbapenemase types were determined by comparing the inhibition zone diameters of the imipenem disc with 3-aminophenylboronic acid (APB) plus ethylenediaminetetraacetic acid (EDTA) to those of the imipenem disc with either APB or EDTA alone, with a breakpoint of 4mm. The overall sensitivities of the current CIT, the modified CIT and NG-Test CARBA 5 were 91.4%, 94.9% and 99.9%, respectively. For detecting isolates co-producing Klebsiella pneumoniae carbapenemase (KPC) and metallo-β-lactamases (MBLs), the modified CIT method had higher sensitivity than the current method (70.0%vs. 53.3%), though this difference was not statistically significant (p=0.063). The NG-Test CARBA 5 showed excellent performance for multi-carbapenemases diagnosis, with sensitivity and specificity of 97.1% and 100%, respectively. Optimizing and standardizing the CIT method for clinical use is necessary. It has certain advantages in diagnosing multi-carbapenemase and rare carbapenemase production. However, for identifying common carbapenemase types, the NG-Test CARBA 5 demonstrated superior performance.

Clinical and Biological Significance of HER2-Low in Ductal Carcinoma In Situ of the Breast

BackgroundDuctal carcinoma in situ (DCIS) is the most common form of preinvasive breast cancer, with 5-10% of cases progressing into invasive disease. Herein, we investigated the association between HER2-low and clinico-pathological characteristics in DCIS and subsequent ipsilateral loco-regional relapse (LRR). Materials and methodsWe accessed our prospectively maintained institutional database. HER2 status was determined by immunohistochemistry and classified as null (score 0), over-expressed (3+), and low (1+ or 2+); in situ hybridization was not considered since it is not used for routine DCIS diagnostics. ResultsAmong 375 patients with DCIS, median age was 54 (27-88) years, with a primary tumor size < 2.5 cm in 63%, grade III in 33%, and positive hormone receptor status (HR) in 81% of cases; 71% underwent breast-conserving surgery, 34% received adjuvant endocrine and 39% radiotherapy. A total of 197 (52%) had tumors with low HER2 expression, which resulted significantly associated with grade I/II (P < .001), Ki67< 20% (P < .001), and HR-positive status (P < .001). HER2-low distribution varied from 19.61% and 50% in ER negative and ER-low (<10%) to 60% and 69% in ER high (50%-95%) and very high tumors (> 95%) (P < .001). After a median 39-month follow-up (IQR 16-65), cumulative incidences of LRR was 0.054. Among 17 patients with paired primary tumor and LRR, 5 had discordant HER2 status, with an even distribution of increased and decreased HER2 expression. ConclusionsLow HER2 expression in DCIS is associated with features of reduced aggressiveness. Importantly, changes in HER2 expression may occur prompting retesting in recurrent cases, in line with observations in invasive breast cancer.

Multiplex PCR for gastrointestinal parasites in stool: Benchmarking against direct microscopy and simplex PCR

PurposeTo develop and validate a multiplex conventional PCR assay to simultaneously detect Cryptosporidium spp., Entamoeba histolytica, and Giardia lamblia in diarrheal samples as a rapid, cost-effective, and sensitive diagnostic tool for prevalent co-infections for improved diagnostic accuracy and efficiency in resource-limited settings. MethodsStool samples collected from patients with gastrointestinal symptoms after taking written consent, processed via wet mount, iodine mount, and PCR assays. Cohen's kappa statistical analysis was done to test agreement. ResultAmong 240 patients, 28.75% showed intestinal protozoa via Microscopy; Single-plex and multiplex PCR demonstrated 100% concordance, detecting 27.9%; confirmed by sequencing. Highest parasite positivity was observed in transplant and immunocompromised patients, with moderate to almost perfect agreement between microscopy and molecular methods. ConclusionMultiplex-conventional PCR offers superior sensitivity and specificity over microscopy and 100% concordance with single-plex PCR, enabling rapid, cost-effective diagnosis of multiple parasites from single stool sample. Its adoption could revolutionize parasitic infection management in routine diagnostics.