title: Evaluation of a semi-automated reporter phage assay for susceptibility testing of Mycobacterium tuberculosis isolates in South Africa.
abstract: In a prospective study conducted by laboratory technologists in a diagnostic laboratory in Cape Town, South Africa, a semi-automated phage-based antibiotic susceptibility assay was implemented and the performance of the luciferase reporter mycobacteriophage (LRP) system for susceptibility testing of clinical Mycobacterium tuberculosis complex (MTC) isolates against rifampin and isoniazid was evaluated. Two hundred consecutive clinical MGIT cultures of MTC species were included in this study. Antibiotic susceptibility assays were set up manually for the LRP and BACTEC radiometric systems (BACTEC) and read in a plate luminometer and the BACTEC 460 instrument, respectively. Discrepant susceptibility results were resolved by the conventional agar proportion method. Of the 200 secondary cultures prepared for this study, 9 (4.5%) were lost to contamination (LRP 4, BACTEC 1, both 4). All of the remaining 191 cultures underwent susceptibility testing by both methods and the overall agreement between the LRP and BACTEC was 98.4% (rifampin 100%; isoniazid 96.9%). Of the 6 discrepant cultures tested by the agar proportion method, 2 gave results in agreement with the LRP. The sensitivity of the LRP for detection of drug-resistant isolates was 100% for both rifampin (n=9) and isoniazid (n=12). The median turnaround time for susceptibility testing was 2 days with the LRP and 9 days with BACTEC. In conclusion, the semi-automated LRP-based assay offers a rapid and practical approach for accurate susceptibility testing of M. tuberculosis cultures in diagnostic laboratories with limited financial resources, but with competent technologists.
authors.affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. nbanaiee@stanford.edu
authors.forename: Niaz
authors.initials: N
authors.lastname: Banaiee
authors.forename: Vanessa
authors.initials: V
authors.lastname: January
authors.forename: Charmaine
authors.initials: C
authors.lastname: Barthus
authors.forename: Maureen
authors.initials: M
authors.lastname: Lambrick
authors.forename: Denise
authors.initials: D
authors.lastname: Roditi
authors.forename: Marcel A
authors.initials: MA
authors.lastname: Behr
authors.forename: William R
authors.initials: WR
authors.lastname: Jacobs
authors.forename: Lafras M
authors.initials: LM
authors.lastname: Steyn
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D001681:Biological Assay; D017930:Genes, Reporter; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics*; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009160:Mycobacteriophages / Q000235:genetics* / Q000502:physiology; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000821:virology; D011446:Prospective Studies; D012293:Rifampin / Q000494:pharmacology; D013019:South Africa; D014376:Tuberculosis / Q000175:diagnosis*
chemical_list: D000995:Antitubercular Agents; D008156:Luciferases; D007538:Isoniazid; D012293:Rifampin