C H A P T E R V I I I
Methods for Bacteriophage Typing
of Mycobacteria
WILLIAM B. REDMOND
Veterans Administration Hospital and Emory University School of Medicine,
Atlanta, Georgia 30322, U.S.A.
JOSEPH H. BATES
Veterans Administration Hospital and University of Arkansas School of Medicine,
Little Rock, Arkansas 72201, U.S.A.
H. W. B. ENGEL
Rzjks Instituut Voor de Volksgezondheid, Bilthoven, Netherlands
I. Introduction .
. 346
A. Early investigations
.
. 346
B. Stimulus by WHO.
.
. 346
11. Media .
. 348
A. RVA-17 agar medium .
. 349
B. RVB-10 liquid medium. Nutrient broth .
. 350
C. Nutrient broth
.
. 351
D. Stock solutions and mixtures .
. 352
111. Bacteriophage .
. 353
A. Isolation of phages.
.
. 353
B. Phage propagation and suspensions .
. 354
C. Phage assay .
. 355
D. Phage mutants and adaptation to new hosts
.
. 355
IV. Epidemiological Studies on Mycobacteria Using Phage Typing
. 357
V. Preservation and Storage; Shipping .
. 364
A. Bacteriophage
.
. 364
B. Mycobacteria
.
. 367
VI. Phage Typing Procedures Adapted to the Mycobacteria
.
. 367
A. The routine test dilution (RTD)
.
. 367
B. Phage and bacterial suspensions
.
. 368
C. Spotting with phage suspensions
.
. 368
D. Reading and reporting of results
.
. 369
E. Methods for subtyping M. tuberculosis
.
. 369
F. The soft-overlay method for typing M. tuberculosis .
. 371
G. Reproducibility of phage typing results
.
. 372
References
.
. 373