Mycoplasma pneumoniae(MP) is a leading pathogen of respiratory infection, especially community-acquired pneumonia (CAP), in children worldwide. However, its diagnosis is frequently ineffective because bacterial culture and serology test are usually positive 1-2 weeks or more after the disease onset. To achieve a better detection efficiency, the single-walled carbon nanotubes(SWCNT) were coupled with the colloidal gold-monoclonal antibody immunochromatographic strips(CGIC). Interestingly, the SWCNT/CGIC assay allowed MP identification, with a detection limit of 1 × 102 copies/ml. Using referenced throat swabs of 97 MP and 40 non-MP cases, the assay yielded 72.2% sensitivity, 100.0% specificity, 100.0% positive predictive value (PPV), 59.7% negative predictive value (NPV). In summary, our assay was far more effective than any conventional methods for the diagnosis of acute MP. The ease of use, rapid and stability further enhance its feasibility for clinical use on-site.