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Journal of Clinical Microbiology, May 2004, p. 2115-2120, Vol. 42, No. 5
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.5.2115-2120.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a Bacteriophage Phage Replication Assay for Diagnosis of Pulmonary Tuberculosis

Ruth McNerney,1* Bupe S. Kambashi,2 Juliana Kinkese,3 Ruth Tembwe,3 and Peter Godfrey-Faussett1

London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom,1 ZAMBART Project, University Teaching Hospital,2 Chest Diseases Laboratory, Ministry of Health, Lusaka, Zambia3

Received 23 June 2003/ Returned for modification 10 November 2003/ Accepted 24 January 2004

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.

* Corresponding author. Mailing address: London School of Hygiene and Tropical Medicine, Keppel St., London WC1E 7HT, United Kingdom. Phone: 44-207-927-2468. Fax: 44-207-637-4314. E-mail: Ruth.Mcnerney{at}lshtm.ac.uk.

Journal of Clinical Microbiology, May 2004, p. 2115-2120, Vol. 42, No. 5
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.5.2115-2120.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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