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There are several problems associated with antimicrobial susceptibility testing (AST) of Haemophilus influenzae. β-Lactamase-negative ampicillin-resistant H. influenzae (BLNAR) isolates with minimum inhibitory concentration (MIC) of ampicillin (ABPC) less then 4 mg/l will be classified as susceptible according to the MIC breakpoint of the CLSI M100 criteria, in spite of harboring penicillin-binding protein (PBP) mutations that cause ABPC resistance. A total of 103 isolates were collected from clinical materials for analysis. The genotypes