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Haemophilus influenzae  is a major human pathogenic bacterium, resulting in a series of diseases, such as pneumonia, bacteremia, meningitis. However, it is hard to diagnose  H .  influenzae  quickly. In this study, the multiple cross displacement amplification (MCDA) and nanoparticle-based lateral flow biosensor (LFB) (MCDA-LFB) were combined to detect  H .  influenzae , which has been proven to be reliable, rapid, and not complicated. On the basis of  H .  influenzae  outer membrane protein  P6  gene, 10 specific primers were designed. The best MCDA condition was 61°C for 1 h. The sensitivity of  H .  influenzae -MCD-LFB assay showed, in the pure cultures, the minimum concentration of genomic DNA templates was 100 fg. The specificity of  H .  influenzae -MCD-LFB assay showed only  H .  influenzae  templates were detected, and no cross-reactivity was found in non- H .  influenzae  isolates and other  Haemophilus  species. In 56 sputum samples, with MCDA-LFB method and PCR detection, 21 samples were positive, which was in consistent with the traditional culture method. The accuracy of diagnosis of MCDA-LFB, in comparison with the traditional culture method and PCR detection, can reach 100%, indicating that the MCDA-LFB assay gains an advantage over the cultured-based method for target pathogen detection. In conclusion, the MCDA-LFB assay is suitable for the sensitive, rapid, and specific detection of  H .  influenzae , which might be used as a potential diagnostic tool for  H .  influenzae  in basic and clinical laboratories.

A Rapid Detection of Haemophilus influenzae Using Multiple Cross Displacement Amplification Linked With Nanoparticle-Based Lateral Flow Biosensor