title: Development of a bacteriophage phage replication assay for diagnosis of pulmonary tuberculosis.
abstract: Successful infection and replication of bacteriophages is indicative of the presence of viable bacteria. We describe here the development of a bacteriophage replication assay for the detection of Mycobacterium tuberculosis by using mycobacteriophage D29. Optimization of phage inoculate and incubation times allowed highly sensitive detection of M. bovis BCG. Fewer than 10 CFU (100 CFU/ml) were detected. No false-positive results were observed in negative samples. Application of the assay to 496 sputum specimens in the National Reference Laboratory of Zambia produced sensitivity, specificity, and positive and negative predictive values of 44.1, 92.6, 82.2, and 67.5%, respectively, compared to culture on Lowenstein-Jensen medium. The equivalent corresponding results for direct fluorescent smear microscopy were 42.3, 96.8, 91.2, and 67.6%. The small increase in sensitivity over that of direct microscopy does not justify the introduction of this technique for routine diagnosis of pulmonary tuberculosis at this time.
authors.affiliation: London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom. Ruth.Mcnerney@lshtm.ac.uk
authors.forename: Ruth
authors.initials: R
authors.lastname: McNerney
authors.forename: Bupe S
authors.initials: BS
authors.lastname: Kambashi
authors.forename: Juliana
authors.initials: J
authors.lastname: Kinkese
authors.forename: Ruth
authors.initials: R
authors.lastname: Tembwe
authors.forename: Peter
authors.initials: P
authors.lastname: Godfrey-Faussett
mesh_terms: D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D015169:Colony Count, Microbial; D006801:Humans; D009160:Mycobacteriophages / Q000502:physiology; D009163:Mycobacterium bovis / Q000302:isolation & purification / Q000821:virology; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification* / Q000821:virology*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000382:microbiology; D014779:Virus Replication; D015024:Zambia