Fluorescent antibody-based detection and ultrastructural analysis of Streptococcus pneumoniae in human sputum DOI Creative Commons

Ana G. Jop Vidal,

Michael J. Francis, Maneesha Chitanvis

et al.

Pneumonia, Journal Year: 2025, Volume and Issue: 17(1)

Published: March 5, 2025

Pneumococcal pneumonia continues to be a significant global health burden, affecting both children and adults. Traditional diagnostic methods for sputum analysis remain challenging. The objective of this study was twofold: develop rapid easy-to-perform assay the identification Streptococcus pneumoniae (Spn) directly in specimens using fluorescence microscopy, characterize with high-resolution confocal microscopy ultrastructure pneumococci residing human sputum. We fluorescently labeled antibodies against pneumococcal capsule (Spn-FLUO). specificity sensitivity Spn-FLUO detecting Spn evaluated vitro vivo mouse models carriage disease, nasopharyngeal specimens, from patients pneumonia. confirmed culture species-specific qPCR assay. strains were serotyped by Quellung. Confocal Imaris software utilized resolve Compared cultures qPCR, demonstrated high (78-96%) samples mice humans. limit detection (LOD) ≥ 1.6 × 10⁴ GenEq/ml. 100%. In lung infected pneumococci, reached 100% LOD 1.39 sputum, 92.7% 3.6 10³ Ultrastructural studies revealed that are expectorated as large aggregates median size 1336 μm². is sensitive within 30 min, encompassing range vaccine non-vaccine serotypes associated highlights most form

Language: Английский

Fluorescent antibody-based detection and ultrastructural analysis of Streptococcus pneumoniae in human sputum DOI Creative Commons

Ana G. Jop Vidal,

Michael J. Francis, Maneesha Chitanvis

et al.

Pneumonia, Journal Year: 2025, Volume and Issue: 17(1)

Published: March 5, 2025

Pneumococcal pneumonia continues to be a significant global health burden, affecting both children and adults. Traditional diagnostic methods for sputum analysis remain challenging. The objective of this study was twofold: develop rapid easy-to-perform assay the identification Streptococcus pneumoniae (Spn) directly in specimens using fluorescence microscopy, characterize with high-resolution confocal microscopy ultrastructure pneumococci residing human sputum. We fluorescently labeled antibodies against pneumococcal capsule (Spn-FLUO). specificity sensitivity Spn-FLUO detecting Spn evaluated vitro vivo mouse models carriage disease, nasopharyngeal specimens, from patients pneumonia. confirmed culture species-specific qPCR assay. strains were serotyped by Quellung. Confocal Imaris software utilized resolve Compared cultures qPCR, demonstrated high (78-96%) samples mice humans. limit detection (LOD) ≥ 1.6 × 10⁴ GenEq/ml. 100%. In lung infected pneumococci, reached 100% LOD 1.39 sputum, 92.7% 3.6 10³ Ultrastructural studies revealed that are expectorated as large aggregates median size 1336 μm². is sensitive within 30 min, encompassing range vaccine non-vaccine serotypes associated highlights most form

Language: Английский

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