LITERATURE UPDATE
this age group, and has raised global health concern. Therefore, there is a pressing need for a rapid, low cost, and user-friendly method for the early diagnosis of M. pneumoniae pneumonia. Herein, the authors develop a
CRISPR sensing platform for on-site M. pneumoniae detection, termed CRAFT (Crispr-based rapid assay device for field testing). The CRAFT provides ‘sample in-result out’ functionality. It completed sample processing and nucleic acid extraction within five minutes at room temperature, with efficiency comparable to commercial kits. RPA and CRISPR/Cas12a reagents were
isolated in a closed tube using a movable magnetic bead valve, and the RPA product was then mixed with the CRISPR reagent. The limit of detection for M. pneumoniae using CRAFT was 100 copies/μL, and the method exhibited no cross-reactivity with other respiratory pathogens. CRAFT was utilised to validate 50 clinical samples, and the results demonstrated 100% consistency with those obtained by qPCR. This versatile platform holds significant
potential for point-of-care testing of M. pneumoniae, particularly in resource- limited setings.
Evaluation of non-invasive diagnostic tests for Mycoplasma pneumoniae pneumonia Fan Y, Tan Z, Wang Z et al. Front Immunol. 2025 Sep 30;16:1656192. doi: 10.3389/fimmu.2025.1656192. eCollection 2025.
Accurate and timely identification of Mycoplasma pneumoniae pneumonia (MPP) remains a clinical challenge. Although nasopharyngeal swab nucleic acid testing (NAAT) and serum IgM antibody assays are widely used, their diagnostic performance varies across studies. This study aimed to retrospectively evaluate the sensitivity and specificity of the two non-invasive methods (NAAT and serum IgM antibody assays) for MPP in real-world clinical setings.
The authors conducted a retrospective
study of adult patients hospitalised for community-acquired pneumonia (CAP) from January 2024 to October 2024. All enrolled patients underwent bronchoalveolar lavage fluid metagenomic next-generation sequencing (BALF- mNGS) and had received at least one of two non-invasive tests (NAAT or serum IgM antibody assays). The sensitivity and specificity of NAAT and serum IgM antibody assays were calculated against the final diagnosis. A non-inferiority test was used to determine whether the sensitivity of NAAT or serum IgM antibody
Three-dimensional computer-generated image of a group of M. pneumoniae (based on a scanning electron microscopic [SEM] image).
assays was not inferior to that of mNGS. Among 594 patients included in
the analysis, 60 were diagnosed with MPP based on a composite reference standard that included laboratory testing results and adjudication by two senior clinicians in accordance with clinical and radiological findings. The sensitivity and specificity of NAAT were 74.1% and 99.3%, respectively, while those of serum IgM antibody assays were 23.6% and 98.0%. McNemar’s test revealed a statistically significant difference in sensitivity between mNGS and the two non-invasive tests (NAAT and serum IgM antibody assays) (P<0.05). The non-inferiority analysis revealed that both NAAT (sensitivity difference: –24.2%, 95% CI: –36.1 to –12.1%; P<0.01) and serum IgM antibody assays (–76.5%, 95% CI: –96.6 to –56.3%; P<0.01) failed to meet the 10% non- inferiority margin compared to mNGS. In clinical practice, a positive result
from either NAAT or serum IgM antibody assays can serve as reliable adjunct evidence for diagnosing MPP. However, in cases with a high clinical suspicion of MPP, negative results from both methods are not sufficient to rule out the diagnosis. For MPP, mNGS remains the most effective diagnostic method compared to non- invasive testing alternatives.
Integrated microfluidic cartridge for rapid and colorimetric detection of Mycoplasma pneumoniae Cui J, Shi B, Chai M et al. Mikrochim Acta.
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WWW.PATHOLOGYINPRACTICE.COM August 2026
2025 Aug 6;192(9):559. doi: 10.1007/s00604-025-07377-6.
Mycoplasma pneumoniae is a major cause of community-acquired pneumonia, yet its diagnosis remains challenging due to overlapping symptoms with other respiratory pathogens. In this study, an integrated microfluidic
cartridge platform has been developed that combines three synergistic advancements: a chemical lysis reagent (GuHCl/EDTA) enabling direct genomic DNA release from M. pneumoniae collected by nasal swabs within two minutes without extraction steps, a novel dual-dye system (hydroxy naphthol blue/SYBR Green I) that leverages complementary fluorescence transitions from red to green for enhanced visual discrimination of loop-mediated isothermal amplification (LAMP) results, and a smartphone-based RGB quantification algorithm to standardise interpretation by analysing colorimetric changes. The microfluidic cartridge integrates
lysis, amplification, and detection into a closed workflow, achieving a sample- to-answer time of 40 minutes with a detection limit of 1 copy/μL. Clinical validation using 25 nasal swab samples demonstrated 95% sensitivity (19/20) and 100% specificity (5/5) compared with serological tests. This platform offers a rapid, cost-effective, and user-friendly solution for point-of-care diagnosis of M. pneumoniae, particularly in resource- limited setings.
PPi
CDC/Sarah Bailey Cutchin/Dan Higgins
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