Mycobacterium tuberculosis DNA detection in soluble fraction of urine from pulmonary tuberculosis patients.

SETTING A tertiary care and research institution in Italy. BACKGROUND Small DNA fragments from cells dying throughout the body have been detected in urine (transrenal DNA [Tr-DNA]). OBJECTIVE To test the hypothesis that Mycobacterium tuberculosis Tr-DNA could be detected in the urine of pulmonary tuberculosis (TB) patients. DESIGN We studied 43 patients with culture-confirmed pulmonary TB with no evidence of extra-pulmonary involvement, 10 patients with pulmonary diseases other than TB and 13 healthy controls. DNA was extracted from urine and analysed by semi-nested polymerase chain reaction (PCR). RESULTS M. tuberculosis-specific sequences were found in the urine of 34 of 43 (79%) TB patients studied, whereas all controls were negative. The transrenal nature of M. tuberculosis DNA was demonstrated by two lines of evidence: first, separate analysis of supernatants and sediments from eight of the study patients found seven positive supernatants but only two matched positive sediments. Second, M. tuberculosis-specific sequences were amplified by semi-nested PCR with primers designed for short but not large amplicons. CONCLUSION Small M. tuberculosis DNA fragments may be detected in the urine of a significant proportion of patients with pulmonary TB. If these observations are confirmed by larger studies, Tr-DNA technology could represent a new approach for detecting pulmonary M. tuberculosis infection.

[1]  S. Umansky,et al.  Transrenal DNA testing: progress and perspectives , 2006, Expert review of molecular diagnostics.

[2]  Denis Boudreau,et al.  Direct molecular detection of nucleic acids by fluorescence signal amplification. , 2005, Journal of the American Chemical Society.

[3]  S. Zanetti,et al.  PCR-based detection of the Mycobacterium tuberculosis complex in urine of HIV-infected and uninfected pulmonary and extrapulmonary tuberculosis patients in Burkina Faso. , 2005, Journal of medical microbiology.

[4]  O. Landt,et al.  Transrenal DNA as a Diagnostic Tool: Important Technical Notes , 2004, Annals of the New York Academy of Sciences.

[5]  P. Stieber,et al.  Therapy Control in Oncology by Circulating Nucleosomes , 2004, Annals of the New York Academy of Sciences.

[6]  Viswanadham Garimella,et al.  Gold nanoparticle-based detection of genomic DNA targets on microarrays using a novel optical detection system. , 2004, Biosensors & bioelectronics.

[7]  D. Maher,et al.  Scale up: meeting targets in global tuberculosis control , 2004, The Lancet.

[8]  P. V. Helden,et al.  Polymerase chain reaction in the diagnosis of urinary tract tuberculosis , 2004, Urological Research.

[9]  C. Piersimoni,et al.  Relevance of Commercial Amplification Methods for Direct Detection of Mycobacterium tuberculosis Complex in Clinical Samples , 2003, Journal of Clinical Microbiology.

[10]  Christopher Dye,et al.  Tuberculosis epidemics driven by HIV: is prevention better than cure? , 2003, AIDS.

[11]  F. Drobniewski,et al.  For Personal Use. Only Reproduce with Permission from the Lancet Publishing Group. Modern Laboratory Diagnosis of Tuberculosis , 2022 .

[12]  Alimuddin Zumla,et al.  Evaluation of PCR-based methods for the diagnosis of tuberculosis by identification of mycobacterial DNA in urine samples. , 2002, The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease.

[13]  S. Umansky,et al.  Circulating Nucleic Acids and Apoptosis , 2001, Annals of the New York Academy of Sciences.

[14]  S. Ni,et al.  Induced apoptosis supports spread of adenovirus vectors in tumors. , 2001, Human gene therapy.

[15]  Arturo Zychlinsky,et al.  Pathogen‐induced apoptosis of macrophages: a common end for different pathogenic strategies , 2000, Cellular microbiology.

[16]  M. El-far,et al.  Rapid diagnosis of genitourinary tuberculosis by polymerase chain reaction and non-radioactive DNA hybridization. , 2000, The Journal of urology.

[17]  A. Garin,et al.  Genetic analysis of DNA excreted in urine: a new approach for detecting specific genomic DNA sequences from cells dying in an organism. , 2000, Clinical chemistry.

[18]  S. Zanetti,et al.  Identification of HIV patients with active pulmonary tuberculosis using urine based polymerase chain reaction assay , 1999, Thorax.

[19]  M. Piacentini,et al.  APOPTOSIS OF HUMAN MONOCYTES/MACROPHAGES IN MYCOBACTERIUM TUBERCULOSIS INFECTION , 1997, The Journal of pathology.

[20]  J. Bates,et al.  Specificity of IS6110-based amplification assays for Mycobacterium tuberculosis complex , 1996, Journal of clinical microbiology.

[21]  A. Friedlander DNA release as a direct measure of microbial killing by phagocytes , 1978, Infection and immunity.