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[This corrects the article DOI: 10.3389/fcimb.2025.1645965.].

Loop-mediated isothermal amplification (LAMP) method is useful for rapidly detecting Edwardsiella ictalurid infection, especially enteric septicemia of catfish (ESC). This study aims to investigate the LAMP method for E. ictaluri detection in ESC. This research was an experimental study using a total of 55 catfish, consisting of 5 negative controls and 50 catfish injected intraperitoneally with 0.1 mL of E. ictaluri at a concentration of 105 CFU/mL. The kidneys of three fishes were randomly selected 6, 12, 18, 24, 30, 36, 42, 48, and 54 hours after infection with E. ictaluri. In addition, samples were also collected on days 3, 5, and 7. Bacterial analysis was determined by conventional biochemistry (genus and species test), whole-genome sequencing of catfish, and LAMP amplification. All types of data obtained in this study were analyzed by descriptive statistics to compare infected and healthy catfish. The catfish were infected with E. ictaluri after 12 hours of infection according to the LAMP amplification procedure which revealed hemorrhages throughout the body, fins, protruding eyes, necrosis, inflammation of the spleen, liver, intestine, pale gills, gut, abdomen swelling, and tissue necrosis in the upper part of the head. It can be concluded that the LAMP method is more effective than the PCR method for detecting infection with E. ictaluri in catfish.

Human Pseudoterranova decipiens larval infections were diagnosed by molecular analysis of mitochondrial cox1 and nd1 genes in 12 health check-up patients in South Korea during 2002-2020. Based on high genetic identity (99.3%-100% for cox1 and 96.7%-98.0% for nd1), we identified all 12 larvae as P. decipiens sensu stricto.

Periprosthetic joint infection (PJI) is a catastrophic complication following joint replacement surgery, posing significant challenges to orthopedic surgeons. Due to the lack of a definitive diagnostic gold standard, timely treatment initiation is problematic, resulting in substantial economic burdens on patients and society. In this review, we thoroughly analyze the complexities of PJI and emphasize the importance of accurate diagnosis and effective treatment. The article specifically focuses on the advancements in diagnostic techniques, ranging from traditional pathogen culture to advanced molecular diagnostics, and discusses their role in enhancing diagnostic accuracy. Additionally, we review the latest surgical management strategies, including everything from debridement to revision surgeries. Our summary aims to provide practical information for the diagnosis and treatment of PJI and encourages further research to improve diagnostic accuracy and treatment outcomes.

ObjectiveBacterial peritonitis (BP) is a serious complication commonly associated with cirrhosis and ascites, often leading to high mortality rates. Although these effects could be reduced with timely and appropriate antibiotics, traditional BP diagnosis relies on culture, often delaying targeted treatment. Therefore, the use of fast molecular assays holds the potential to enhance laboratory diagnosis. In this study, we assessed the diagnostic performaance of Molecular Culture ID, a broad PCR-based assay targeting the 16S-23S interspace rDNA region in the scope of BP diagnosis.MethodsThe residual material from 247 peritoneal fluid samples submitted for routine diagnostics was analyzed using Molecular Culture ID and compared alongside the standard of care (SOC) results.ResultsSample positivity and species identification outcomes of Molecular Culture ID were compared to those of SOC. Molecular Culture ID yielded 1.6x more positive samples than SOC. Percent positive agreement (PPA) between Molecular Culture ID and SOC at the sample level was 90.1% (IC 95%, 81.0% to 95.1%), and negative percent agreement (NPA) was 70.5% (IC 95%, 63.3% to 76.7%). At the species level, the PPA was 75.2% (95% CI 67.2% to 81.8%). Molecular Culture ID yielded 289 extra bacterial identifications, mainly anaerobic species. High leukocyte counts, indicative of infection, were concordant with Molecular Culture ID positivity.ConclusionMolecular Culture ID demonstrated enhanced BP diagnostic capabilities compared to SOC, with higher positivity rates, more comprehensive species identification for difficult to culture species and a high correlation with leukocyte counts.

Efficient, wide-scale testing for SARS-CoV-2 is crucial for monitoring the incidence of the infection in the community. The gold standard for COVID-19 diagnosis is the molecular analysis of epithelial secretions from the upper respiratory system captured by nasopharyngeal (NP) or oropharyngeal swabs. Given the ease of collection, saliva has been proposed as a possible substitute to support testing at the population level. Here, we used a novel saliva collection device designed to favour the safe and correct acquisition of the sample, as well as the processivity of the downstream molecular analysis. We tested 1003 nasopharyngeal swabs and paired saliva samples self-collected by individuals recruited at a public drive-through testing facility. An overall moderate concordance (68%) between the two tests was found, with evidence that neither system can diagnose the infection in 100% of the cases. While the two methods performed equally well in symptomatic individuals, their discordance was mainly restricted to samples from convalescent subjects. The saliva test was at least as effective as NP swabs in asymptomatic individuals recruited for contact tracing. Our study describes a testing strategy of self-collected saliva samples, which is reliable for wide-scale COVID-19 screening in the community and is particularly effective for contact tracing.

We present a case of acute clonal bone marrow 98% infiltration of atypical myeloid cells with borderline hypogranular/agranular promyelocytes/myelocytes and occasional blast cells maturity, which also formed extramedullary tumours in the chest wall, with isolated trisomy of chromosome 6 and pathogenic variant U2AF1 (S34F) that escapes established acute myeloid leukaemia (AML) diagnostic criteria according to the World Health Organization (WHO) classification. Following standard daunorubicin and cytarabine induction therapy, the disease progressed with the appearance of a previously undetected clone of leukaemic cells with a distinct immunophenotype demonstrating monocytoid differentiation and clonal evolution to a hypo‐tetraploid karyotype with an average number of 84 chromosomes and new pathogenic NRAS and ZRSR2 mutations. The patient reactivated refractory disseminated intravascular coagulation (DIC) leading to a progressive supratentorial hematoma and finally cardiac arrest. In conclusion, our report shows that atypical clonal myelocytes can massively infiltrate the bone marrow and form extramedullary tumours, justifying the diagnosis and treatment of acute leukaemia, although they did not fit the current classification.

Improved resolution of molecular diagnostic technologies enabled detection of smaller sized exonic level copy-number variants (CNVs). The contribution of CNVs to autosomal recessive (AR) conditions may be better recognized using a large clinical cohort. We retrospectively investigated the CNVs’ contribution to AR conditions in cases subjected to chromosomal microarray analysis (CMA, N=~70,000) and/or clinical exome sequencing (ES, N=~12,000) at Baylor Genetics; most had pediatric onset neurodevelopmental disorders. CNVs contributed to biallelic variations in 87 cases, including 81 singletons and three affected sibling pairs. Seventy cases had CNVs affecting both alleles, and 17 had a CNV and a single-nucleotide variant (SNV)/indel in trans. In total, 94.3% of AR-CNVs affected one gene; among these 41.4% were single-exon and 35.0% were multiexon partial-gene events. Sixty-nine percent of homozygous AR-CNVs were embedded in homozygous genomic intervals. Five cases had large deletions unmasking an SNV/indel on the intact allele for a recessive condition, resulting in multiple molecular diagnoses. AR-CNVs are often smaller in size, transmitted through generations, and underrecognized due to limitations in clinical CNV detection methods. Our findings from a large clinical cohort emphasized integrated CNV and SNV/indel analyses for precise clinical and molecular diagnosis especially in the context of genomic disorders.

Avian influenza viruses of the H9 subtype cause significant losses to poultry production in endemic regions of Asia, Africa and the Middle East and pose a risk to human health. The availability of reliable and updated diagnostic tools for H9 surveillance is thus paramount to ensure the prompt identification of this subtype. The genetic variability of H9 represents a challenge for molecular-based diagnostic methods and was the cause for suboptimal detection and false negatives during routine diagnostic monitoring. Starting from a dataset of sequences related to viruses of different origins and clades (Y439, Y280, G1), a bioinformatics workflow was optimized to extract relevant sequence data preparatory for oligonucleotides design. Analytical and diagnostic performances were assessed according to the OIE standards. To facilitate assay deployment, amplification conditions were optimized with different nucleic extraction systems and amplification kits. Performance of the new real-time RT-PCR was also evaluated in comparison to existing H9-detection methods, highlighting a significant improvement of sensitivity and inclusivity, in particular for G1 viruses. Data obtained suggest that the new assay has the potential to be employed under different settings and geographic areas for a sensitive detection of H9 viruses.

Background & objectives: Dengue (DEN) is a result of infection by one or multiple types of four dengue viruses known as Dengue virus (DENV) 1-4. Identifying circulating serotype and genotype is epidemiologically important, however, it is challenging in resource limited areas. Moreover, transporting samples from the collation site to the laboratory in appropriate condition is an exigent task. To overcome this, we evaluated the usefulness of dry blots of serum for DENV diagnosis, serotyping and genotyping. Methods: Serum samples received for diagnosis were divided into parts; one was used for providing the diagnosis. Remaining sample was distributed in three parts (100 µl each), one part was used for molecular testing and two parts were mixed with RNAlater reagent® in equal volumes and was blotted on Whatman filter paper no 3. The blots were dried and stored at 4°C and 28°C and tested for presence of dengue RNA, serotypes and genotypes after 7 days of incubation. Results: The diagnosis and serotyping results of serum sample and dry serum blots were in concordance. Out of 20 positive samples, 13 (65%) gave satisfactory sequencing results. Genotype III of DENV-1, Genotype IV of DENV 2 and Genotype I of DENV-4 were detected. Interpretation & conclusion: The results demonstrate that serum mixed with RNA protective solution and blotted on Whatman filter paper no 3 can be effectively used for diagnosis, serotyping and genotyping of DENVs. This will help in easy transportation, diagnosis and effective data generation in resource limited settings.