The diagnosis of neurogenetic disorders is often prolonged due to clinical variability and significant genetic heterogeneity, making molecular diagnosis challenging. This study aimed to investigate the molecular basis of rare inherited neurological conditions in three consanguineous Moroccan families. We performed Whole exome sequencing (WES) on probands from three unrelated consanguineous Moroccan families (NP 69, NP 84, NP 89). Candidate variants were validated by Sanger sequencing and segregated within the families. Pathogenicity was assessed through in silico prediction tools. Molecular modeling and molecular dynamics simulations were applied to assess the structural impact of selected variants. Whole exome sequencing revealed likely disease-causing variants in all three families. In Family NP 69, we found compound heterozygous missense variants in RYR3 gene (p.Gly2168Arg and p.Val854Ile) in a proband presenting with developmental delay and hippocampal sclerosis. In Family NP 84, a homozygous missense variant (p.Trp671Arg) in POLR3A gene, was associated with classic hypomyelinating leukodystrophy and ataxia, but with atypical features including preserved cognition, myoclonus, and oligodontia, consistent with recurrence of a previously reported variant in North Africa and suggestive of a possible founder effect. In Family NP 89, a homozygous splice-site mutation in LAMA2 gene (c.8244 + 1G > A) was confirmed in a proband with severe congenital muscular dystrophy. This study reports a novel compound heterozygous configuration in RYR3, confirms a previously reported bi-allelic variant in POLR3A, and validates a recurrent LAMA2 variant. Together, these findings add new insights into the genotype–phenotype spectrum of neurogenetic disorders in consanguineous North African populations, all consistent with a shared autosomal recessive inheritance pattern.
Tuberculosis (TB) is a major health burden, the leading cause of death among people living with HIV, and one of the main causes of death linked to antimicrobial resistance. The World Health Organization (WHO) launched in 2015 the End TB Initiative that aims to reduce TB cases and deaths by 2035. However, despite some progress, significant gaps persist in TB detection, treatment, prevention, and funding. Diagnosis is most often by sputum smear and culture and, when available, by nucleic acid amplification tests (NAATs), which revolutionizes TB diagnosis and monitoring, improving early detection, resistance surveillance, and treatment response evaluation. In this review, we report on the implication of various Real-Time PCR (RT-PCR) laboratory diagnostic tests and compare their performance in detecting MTB. We show that the Xpert MTB/RIF test is currently the most effective and widely validated molecular tool for diagnosing pulmonary TB, though its accuracy in extrapulmonary TB remains inconsistent. These findings suggest that combining molecular tests with other methods—such as radiology, microbiology, biomarkers, and immunological approaches—could provide a more reliable diagnostic strategy. Scaling up molecular tools worldwide, especially in high-burden regions, is essential to achieving the WHO End TB Strategy and ultimately eliminating TB.
The COVID-19 pandemic highlighted critical gaps in serological diagnostic capacity in low- and middle-income countries, particularly the strategic dependence on imported commercial kits. Here we report the development, optimisation, and analytical validation of an in-house quantitative indirect enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of IgG antibodies against the SARS-CoV-2 Spike protein (S1-S2). Four antigen coating concentrations (1, 2, 3 and 5 µg/mL) and three serum dilutions (1:50, 1:100 and 1:1000) were systematically evaluated. An internal calibration standard was prepared from a pool of six convalescent sera. Analytical validation followed CLSI EP17-A criteria and included linearity assessment, intra- and inter-assay precision (coefficient of variation, CV%), and analytical limits (limit of blank [LOB], limit of detection [LOD], and limit of quantification [LOQ]). Preliminary diagnostic performance was evaluated on an independent pilot cohort of 20 sera (10 RT-PCR-confirmed COVID-19, 10 pre-pandemic). The optimal conditions were antigen coating at 2 µg/mL and serum dilution 1:100. The calibration curve showed strong linearity (R² = 0.996; y = 0.0050x + 0.0793). Intra-assay CVs ranged from 4.1% to 6.6% and inter-assay CVs from 2.7% to 8.8%, both within acceptance criteria (≤10% and ≤15%, respectively). All analytical limits were well below positive signals. On the pilot cohort, the assay achieved sensitivity of 90.0% (95% CI: 59.6–98.2%), specificity of 90.0% (95% CI: 59.6–98.2%), and diagnostic accuracy of 90.0% (95% CI: 69.9–97.2%), with an area under the receiver operating characteristic curve (AUC-ROC) of 0.92. These preliminary results suggest the feasibility of locally developing a quantitative ELISA platform applicable to serological surveillance of SARS-CoV-2 and, more broadly, to future in-house immunoassay development for emerging infectious diseases.
Prostate cancer (PCa) has emerged as the leading malignancy among Moroccan men. This brief communication analyses recent data (2018–2025) to outline the epidemiological transition of PCa in Morocco, characterised by a 77% surge in incidence over a 30-year trajectory. Despite improved diagnostic capabilities, the mortality-to-incidence ratio (MIR) (0.49) remains significantly higher than in high-Human Development Index (HDI) countries (< 0.20). This analysis highlights the urgent need for structured early-detection strategies to address the crisis of late-stage diagnosis in the North African context.
Differential gene expression analysis of RNA Sequencing (RNA-Seq) data is crucial for understanding key patterns of gene regulation and enhancing our knowledge of biological processes and diseases. The workflow of this analysis comprises quality control, filtering of low-quality data, alignment, read counting, and final differential analysis. In this case, users often need to manually combine several tools and write multiple scripts to cover the entire pipeline. This fragmented approach is time-consuming and not user-friendly, especially for non-expert users. There is a need for an integrated, automated and accessible solution that unifies the entire analysis process within a single, easy-to-use platform. To address this need, we developed SeqExpressionAnalyser, an R package that provides a web application for interactive differential gene-expression analysis of RNA-seq data, making it accessible to R users for the first time. Built on the Shiny framework, SeqExpressionAnalyser enables users to read FASTQ files and perform analyses, including quality control, filtering, alignment, read counting, and differential expression analysis. The tool generates multiple outputs, including data tables, an HTML report and visualisations. The source code is available on GitHub (https://github.com/sanaeesskhayry/SeqExpressionAnalyser) and is licensed under the GPLv3 license. Also available as a Docker image at https://hub.docker.com/repository/docker/biomix/seq-expression-analyser/general.
