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 cap b 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.
Keywords: group III-like mutations, BLPACR, prevalence, invasive Haemophilus influenzae , genome