Five tests--satellitism, synthesis of porphyrins, acid production from sucrose, beta-galactosidase activity (ONPG), and indole production--to differentiate between strains of Haemophilus influenzae and strains of V-dependent Haemophilus species were evaluated. Six per cent of strains of H influenzae were misidentified as H parainfluenzae by a test for satellitism using filter paper discs impregnated with X factor, V factor, or both, applied to Columbia Agar. None of seven nutrient agars tested grew Haemophilus species, and determined accurately the X factor requirement. Synthesis of porphyrins from delta-aminolaevulinic acid provided a reliable means of demonstrating that X factor was required. A test for the production of acid from sucrose discriminated successfully between strains of V-dependent Haemophilus species (positive) and H influenzae (negative). Most isolates were identified correctly by the ONPG test, but occasional V-dependent strains were negative and could be misidentified as H influenzae. The discriminative value of the indole test was unsatisfactorily low. The results of the tests are discussed in relation to the identification of H influenzae in the diagnostic laboratory.
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