An IMS/ATP Assay for the Detection of Mycobacterium tuberculosis in Urine.
Background. Although sputum smears are the gold standard for diagnosis of tuberculosis, sensitivity in HIV/TB coinfection cases is low, indicating a need for alternative methods. Urine is being increasingly evaluated. Materials and Methods. A novel method for detecting Mycobacterium tuberculosis (MTB) in synthetic urine using a combined IMS/ATP assay was evaluated. Preliminary work established standard ATP conditions and the sensitivity and specificity of the MTB antibody. Eighty-four blinded samples in four replicate assays were evaluated for the presence of MTB using labeled immunomagnetic beads for capture. Beads were separated, washed, and resuspended in broth and added to a microtiter plate. Bioluminescent output was measured and signal-to-noise ratios were calculated. All samples were plated on Middlebrook 7H10 agar or trypticase soy agar to determine limit of detection and recoveries. Results and Conclusions. MTB was distinguished from common bacteriuria isolates and other nontarget bacteria by its ATP results. IMS/ATP successfully detected 19 of 28 samples of MTB in synthetic urine with a limit of detection of 10(4) CFU/ml. Sensitivity and specificity were 67.9% and 82.1%, respectively. This assay offers a possible rapid screening method for HIV-positive patients with suspected coinfection to improve MTB diagnosis.
Department of Cell Biology, Microbiology and Molecular Biology, University of South Florida, 4202 E. Fowler Avenue, ISA 2015, Tampa, FL 33620-7115, USA.
Dawn M
DM
Hunter
Daniel V
DV
Lim