An antigenic analysis of the mycobacteria, Mycobacterium fortuitum, Myco. kansasii, Myco. phlei, Myco. smegmatis and Myco. tuberculosis

Abstract

THE genus Mycobacterium contains a number of fairly clearly defined species, which may be obligatory pathogens, opportunistic pathogens or saprophytes, together with an ill-defined group of anonymous strains. So far, the definition of species has relied largely on morphology, cultural appearances, biochemical activity and temperature requirements. The grouping of strains by Runyon (1959) and Marks and Richards (1962) relies on these and related criteria. Information about the antigenic components of a number of species and unclassified strains has been accumulating over the past few years (Parlett and Youmans, 1958; Castelnuovo et al., 1960; Lind, 1961 ; Sushida and Hirano, 1963; Schaefer, 1965; Jensen, Klaer and Lundberg, 1964, 1966a, b andc). This paper records an antigenic analysis of Mycobacterium fortuitum, Myco. kansasii, Myco. phlei, Myco. smegmatis and Myco. tuberculosis made by Ouchterlony's double diffusion-in-agar precipitation technique. MATERIALS AND METRODS OrganismsAntigen extracts were prepared from 25 strains of mycobacteria that included some obtained from the National Collection of Type Cultures (NCTC) and some isolated by us from patients and soil (PGH). They were: Mycobacterium fortuitum strains NCTC 8573t, 946, 1542,2006,2291,2891,3631 and 8697, and PGH strains " Hoey "t, '' Ugoji " and " Soil "t; Myco. kanrarii strains PGH " Brown "t, " Walker ", " Birk " and " Sheppard "; Myco. phlei strains NCTC 8156t, 525, 8151, 8157, 10008 and 10266; Myco. smegmatis strains NCTC 3337, 8159 and 10265; and Myco. tuberculosis strain H37 Rat. Strains marked with an obelus were also used for preparation of antisera. Owing to the risk involved in the work with living bacteria only one representative of Myco. tuberculosis was used.All the strains of Myco. kansasii and the Hoey strain of Myco. fortuitum were isolated from the sputum of patients. Myco. fortuitum strain Ugoji was recovered from a patient's abscess and Myco. fortuitum strain Soil from a sample of garden soil. All strains of Myco. kansasiiexcept strain Walker, which was a single isolation, were clinically signiscant in that they had repeatedly been found in the patient's sputum. Their identification, and that of the Myco. fortuitum cultures isolated by Present address: (1959, 1965). Preparation of antigensCultures were grown at 37°C in I-litre volumes of modified Sauton's medium (Boyden and Sorkin, 1955) in large Carrel or Thompson flasks, and their pellicles were harvested when no further growth appeared to be taking place. The age of the cultures depended on the species used and varied from 1 to 8 wk. Parts of the harvested pellicles were extracted at -20°C in an " X " press (Edebo, 1961).Microscopical examination of the product obtained after three pressings revealed that the cell walls were almost totally broken up. The crushed material was allowed to stand at 4°C and the cleared fluid layer between the surface flocculate and the sediment was then pipetted off. This extract served as antigen and was stored in small volumes at -20°C.

Studies On Mycobacterium Chelonei

“…All strains were examined by immunodiffusion analysis, reactants being prepared by the methods previously described (Stanford and Beck, 1968;Stanford and Gunthorpe, 1971). Antisera were raised in rabbits to the type strains of Myco.…”

A Co-operative Numerical Analysis of Rapidly Growing Mycobacteria

“…For these studies the soluble portion of niycobacterial lysates was tested in immunodiffusion tests with selected antisera (Stanford & Beck, 1968 ;Kwapinski, Alcasid & Palser, 1970).…”

An Immunodiffusion Analysis of Strains of Mycobacterium Ulcerans Isolated in Australia, Malaya, Mexico, Uganda and Zaire

“…DISCUSS~ON Methods different from those previously described (Stanford and Beck, 1968;Stanford and Gunthorpe, 1971) had to be employed because the study strains grew very poorly on liquid media and hardly at all on simple media. The use of Mudd's (1925) observation of the lipophilic nature of mycobacteria to purify the organisms was necessary because in initial experiments rabbits died during immunisation from anaphylaxis caused by egg protein.…”

“…Imrnunodiffusion tests were carried out as previously described (Stanford and Beck, 1968). Antigens of representative strains from each country were tested with each antiserum and antigens of all strains were tested with the best two antisera (those to NCTC no.…”