High Prevalence of Group III-Like Mutations Among BLPACR and First Report of Haemophilus influenzae ST95 Isolated from Blood in China

PURPOSE: We aimed to evaluate antibiotic resistance and molecular epidemiological characteristics of non-invasive Haemophilus influenzae (H. influenzae) from pneumonia patients and analyze the whole genome of one invasive H. influenzae isolated from blood in pediatric patients. METHODS: Antibiotic susceptibility was tested using the turbidimetric method. β-lactamase-producing and serotyping genes were evaluated via multiplex polymerase chain reaction (PCR), and ftsI was amplified using high-fidelity PCR. Lastly, whole genome sequencing (WGS) was conducted using Illumina HiSeq and PacBio sequencing technology. RESULTS: We observed that the ampicillin (AMP) and amoxicillin/clavulanate (AMC) resistance rates of non-invasive H. influenzae were as high as 99.06% (after adjustment) and 49.53%, respectively. The β-lactamase gene of 106 AMP-resistant strains was bla(TEM-1). Group III-like mutation accounted for 71.15% of β-lactamase-positive, AMC-resistant (BLPACR) strain mutants. The novel Asn-526→His mutation was present in one β-lactamase-negative AMP-susceptible (BLNAS) strain. Non-invasive H. influenzae strains all belonged to non-typeable H. influenzae (NTHi). In contrast, the invasive H. influenzae 108 isolated from blood in China belonged to H. influenzae type b (Hib). It belonged to sequence typing ST95 and exhibited sensitivity to all 11 antibiotics. Three prophages were identified, and the capb loci of the H. influenzae strain 108 revealed regions I–III exist in duplicate; however, complete deletion of IS1016 was only present in one of the copies. CONCLUSION: Non-invasive H. influenzae NTHi with β-lactamase-positive was highly prevalent. Notably, group III-like mutations had increased prevalence among BLPACR strains. H. influenzae belonging to Hib and ST95 was first reported to cause sepsis in China.