The Antigens of Pseudomonas aeruginosa Studied by the Ouchterlony Technique and Immuno-electrophoresis

Eeden, D. Van

doi:10.1099/00221287-48-1-95

Cited by 2 publications

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“…The present experiments are in agreement with the finding of van Eeden (1967) that trichloracetic acid extracts of P. aeruginosa strains exhibited different immunoelectrophoretic patterns. Antigens of van Eeden's four strains running towards the anode correspond to our IE-group I; antigens precipitating on the cathode side are comparable with our group-IV antigens, while those appearing on both sides of the antigen well may belong to our groups I1 and 111.…”

Section: Antigens Of Pseudomonas Aeruginosa 233supporting

confidence: 82%

Classification of Pseudomonas Aeruginosa O Antigens by Immunoelectrophoresis

Lányi¹,

Ádám²,

Szentmihályi³

1975

Journal of Medical Microbiology

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IN a study reported previously (LBnyi and AdBm, 1973), it was shown that double agar-gel precipitation and agglutination give, as a rule, identical crossreaction patterns for the 0 antigens of Pseudomonas aeruginosa. On the basis of precipitation of different kinds of bacterial extract, the antigens were classified into the following groups: (1) those of which saline extracts, supernates of phenol-water extracts (L1 fractions) and purified lipopolysaccharide (LPS) precipitated readily with homologous antisera; (2) those in which saline extracts and LPS precipitated but L1 fractions did not; and (3) two partial antigens, none of the extracts of which precipitated with absorbed antisera, although the presence of the antigen was demonstrable by means of agglutination tests. In the experiments described in this paper, a further classification of P. aeruginosa 0 antigens was attempted. MATERIALS AND METHODSBacterial strains. Twenty-three P. aeruginosa cultures represented Unyi's 0-antigen type strains (Lanyi, 1966-67 and1970); in addition, 53 strains of different serogroups were chosen from our collection. Of 13 strains received from Professor P. V. Liu, Louisville, Kentucky, USA, as type strains of the provisional international P. aeruginosa antigenic scheme, 12 originated from Habs (1957) and one from Sandvik (1960). Saline extracts were made from all these cultures, but only Lanyi's 23 0-antigen type strains were used for the preparation of L1, LPS, trichloracetic-acid (TCA) and alkali extracts.Antigens. Extracts in physiological saline (" saline ") were prepared by heating the bacterial suspension at 100°C for 23 h; each ml of the supernate obtained after centrifugation represented c. 30 mg of bacteria (moist weight). L1 fractions were the freeze-dried supernates of ultracentrifuged phenol-water extracts ; nucleic acid-free LPS preparations were dialysed and freeze-dried phenol-water extracts purified by ultracentrifugation at 105,000 g. L1 and LPS solutions were prepared for precipitation by dissolving 5 mg of the freeze-dried preparation in saline and heating at 100°C for 1 h. The preparation of these antigens was described in detail by Adam, Kontrohr and Horvath (1971) and by Unyi and A d h (1973).The bacteria were, as a rule, grown in Roux flasks at 37°C for 24 h on nutrient-agar medium : beef extract (Central Slaughterhouse, Budapest) 15 g ; peptone (Richter, Budapest) 10 g; NaCl3 g; NazHP04.12H20 4 g; agar 18 g; tap water 10oO ml; pH 7.4. Bloodagar cultures were grown on plates containing baker's yeast 40 g; peptone (Richter, Budapest) 10 g; NaCl3 g; Na2HP04. 12H20 4 g; agar 18 g; tap water 10oO ml; pH 7.4; ox blood 50 ml.TCA extracts were prepared by suspending 5 g of dry bacteria or 10 g of moist bacteria in 50 ml of ice-cold TCA 10% (w/v). The suspension was left to stand in the refrigerator at 4"-6"C for 24-48 h and then centrifuged. After a second extraction of the deposit, the

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Section: Antigens Of Pseudomonas Aeruginosa 233supporting

confidence: 82%