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Published in: Infection 3/2017

01-06-2017 | Original Paper

Detection of transrenal DNA for the diagnosis of pulmonary tuberculosis and treatment monitoring

Authors: Ines Labugger, Jan Heyckendorf, Stefan Dees, Emilia Häussinger, Christian Herzmann, Thomas A. Kohl, Elvira Richter, Eric Rivera-Milla, Christoph Lange

Published in: Infection | Issue 3/2017

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Abstract

Purpose

Molecular diagnostics of patients with MTB tuberculosis from urine samples.

Methods

We developed a new molecular assay based on the detection of M. tuberculosis-specific transrenal DNA (trDNA) and tested it for the diagnosis of active tuberculosis at the initiation of anti-tuberculosis therapy and during treatment follow-up.

Results

The overall sensitivity of trDNA was 96 and 100% when smear-microscopy and trDNA was combined. In a subset of TB treatment naïve patients (n = 11) sensitivity and specificity of trDNA was 64 and 100%, respectively. For this subset of patients the sensitivity was 91% when smear-microscopy and trDNA diagnosis were combined. After treatment initiation, trDNA showed a significant reduction in concentration over time reaching undetectable trDNA values at week 12 in 9 of 11 accessible patients (82%). Kinetics in treatment-naïve patients showed low base-line trDNA levels, which increased to maximal trDNA levels within one week indicating bactericidal activity of anti-tuberculosis drugs after the initiation of effective therapy. Maximal trDNA levels correlated positively with a radiological score, suggesting that the process of DNA excretion may reflect the extent of pulmonary disease. Matched samples showed an inverse correlation between the time to positivity of solid culture with maximum trDNA levels as well as the expected positive correlation between smear grade and maximum trDNA values.

Conclusion

The detection of M. tuberculosis trDNA from urine specimen is a promising method for the diagnosis tuberculosis. The assay may be a candidate diagnostic tool for patients with paucibacillary and extrapulmonary disease, as method to assess treatment responses and could be helpful to diagnose tuberculosis in children.
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Literature
1.
go back to reference World Health Organisation. WHO Global Tuberculosis Report 2015. World Health Organisation. WHO Global Tuberculosis Report 2015.
2.
go back to reference Lonnroth K, Migliori GB, Abubakar I, D’Ambrosio L, de Vries G, Diel R, Douglas P, Falzon D, Ma Gaudreau, Goletti D, Gonzalez Ochoa ER, LoBue P, Matteelli A, Njoo H, Solovic I, Story A, Tayeb T, van der Werf MJ, Weil D, Zellweger JP, Abdel Aziz M, Al Lawati MRM, Aliberti S, Arrazola de Onate W, Barreira D, Bhatia V, Blasi F, Bloom A, Bruchfeld J, Castelli F, Centis R, Chemtob D, Cirillo DM, Colorado A, Dadu A, Dahle UR, De Paoli L, Dias HM, Duarte R, Fattorini L, Gaga M, Getahun H, Glaziou P, Goguadze L, del Granado M, Haas W, Jarvinen A, Kwon GY, Mosca D, Nahid P, et al. Towards tuberculosis elimination: an action framework for low-incidence countries. Eur Respir J. 2015;45:928–52.PubMedPubMedCentral Lonnroth K, Migliori GB, Abubakar I, D’Ambrosio L, de Vries G, Diel R, Douglas P, Falzon D, Ma Gaudreau, Goletti D, Gonzalez Ochoa ER, LoBue P, Matteelli A, Njoo H, Solovic I, Story A, Tayeb T, van der Werf MJ, Weil D, Zellweger JP, Abdel Aziz M, Al Lawati MRM, Aliberti S, Arrazola de Onate W, Barreira D, Bhatia V, Blasi F, Bloom A, Bruchfeld J, Castelli F, Centis R, Chemtob D, Cirillo DM, Colorado A, Dadu A, Dahle UR, De Paoli L, Dias HM, Duarte R, Fattorini L, Gaga M, Getahun H, Glaziou P, Goguadze L, del Granado M, Haas W, Jarvinen A, Kwon GY, Mosca D, Nahid P, et al. Towards tuberculosis elimination: an action framework for low-incidence countries. Eur Respir J. 2015;45:928–52.PubMedPubMedCentral
3.
go back to reference Perkins MD, Cunningham J. Facing the crisis: improving the diagnosis of tuberculosis in the HIV era. J Infect Dis. 2007;196(Suppl 1):S15–27.CrossRefPubMed Perkins MD, Cunningham J. Facing the crisis: improving the diagnosis of tuberculosis in the HIV era. J Infect Dis. 2007;196(Suppl 1):S15–27.CrossRefPubMed
4.
go back to reference Olaru ID, Heyckendorf J, Grossmann S, Lange C. Time to culture positivity and sputum smear microscopy during tuberculosis therapy. PLoS One. 2014;9:e106075.CrossRefPubMedPubMedCentral Olaru ID, Heyckendorf J, Grossmann S, Lange C. Time to culture positivity and sputum smear microscopy during tuberculosis therapy. PLoS One. 2014;9:e106075.CrossRefPubMedPubMedCentral
5.
go back to reference Lawn SD, Kerkhoff AD, Vogt M, Wood R. Diagnostic accuracy of a low-cost, urine antigen, point-of-care screening assay for HIV-associated pulmonary tuberculosis before antiretroviral therapy: a descriptive study. Lancet Infect Dis. 2012;12:201–9.CrossRefPubMedPubMedCentral Lawn SD, Kerkhoff AD, Vogt M, Wood R. Diagnostic accuracy of a low-cost, urine antigen, point-of-care screening assay for HIV-associated pulmonary tuberculosis before antiretroviral therapy: a descriptive study. Lancet Infect Dis. 2012;12:201–9.CrossRefPubMedPubMedCentral
6.
go back to reference Torrea G. PCR-based detection of the Mycobacterium tuberculosis complex in urine of HIV-infected and uninfected pulmonary and extrapulmonary tuberculosis patients in Burkina Faso. J Med Microbiol. 2005;54:39–44.CrossRefPubMed Torrea G. PCR-based detection of the Mycobacterium tuberculosis complex in urine of HIV-infected and uninfected pulmonary and extrapulmonary tuberculosis patients in Burkina Faso. J Med Microbiol. 2005;54:39–44.CrossRefPubMed
7.
go back to reference Kroidl I, Clowes P, Reither K, Mtafya B, Rojas-Ponce G, Ntinginya EN, Kalomo M, Minja LT, Kowuor D, Saathoff E, Kroidl A, Heinrich N, Maboko L, Bates M, O’Grady J, Zumla A, Hoelscher M, Rachow A. Performance of urine lipoarabinomannan assays for paediatric tuberculosis in Tanzania. Eur Respir J. 2015;46:761–70.CrossRefPubMed Kroidl I, Clowes P, Reither K, Mtafya B, Rojas-Ponce G, Ntinginya EN, Kalomo M, Minja LT, Kowuor D, Saathoff E, Kroidl A, Heinrich N, Maboko L, Bates M, O’Grady J, Zumla A, Hoelscher M, Rachow A. Performance of urine lipoarabinomannan assays for paediatric tuberculosis in Tanzania. Eur Respir J. 2015;46:761–70.CrossRefPubMed
9.
go back to reference Luetkemeyer AF, Firnhaber C, Kendall MA, Wu X, Mazurek GH, Benator DA, Arduino R, Fernandez M, Guy E, Johnson P, Metchock B, Sattler F, Telzak E, Wang YF, Weiner M, Swindells S, Sanne IM, Havlir DV, Grinsztejn B, Alland D; AIDS Clinical Trials Group A5295 and Tuberculosis Trials Consortium Study 34 Teams. Evaluation of Xpert MTB/RIF to identify pulmonary tuberculosis in tuberculosis suspects from low and higher prevalence settings compared to acid fast smear and culture. Clin Infect Dis. 2016;62(9):1081–8.CrossRefPubMedPubMedCentral Luetkemeyer AF, Firnhaber C, Kendall MA, Wu X, Mazurek GH, Benator DA, Arduino R, Fernandez M, Guy E, Johnson P, Metchock B, Sattler F, Telzak E, Wang YF, Weiner M, Swindells S, Sanne IM, Havlir DV, Grinsztejn B, Alland D; AIDS Clinical Trials Group A5295 and Tuberculosis Trials Consortium Study 34 Teams. Evaluation of Xpert MTB/RIF to identify pulmonary tuberculosis in tuberculosis suspects from low and higher prevalence settings compared to acid fast smear and culture. Clin Infect Dis. 2016;62(9):1081–8.CrossRefPubMedPubMedCentral
10.
go back to reference DiNardo AR, Lange C, Mandalakas AM. 1, 2, 3 (Years)  …  and you’re out: the end of a 123-year Historic Era. Clin Infect Dis. 2016;62(9):1089–91.CrossRefPubMed DiNardo AR, Lange C, Mandalakas AM. 1, 2, 3 (Years)  …  and you’re out: the end of a 123-year Historic Era. Clin Infect Dis. 2016;62(9):1089–91.CrossRefPubMed
11.
go back to reference Karinja MN, Esterhuizen TM, Friedrich SO, Diacon AH. Sputum volume predicts sputum mycobacterial load during the first 2 weeks of antituberculosis treatment. J Clin Microbiol. 2015;53:1087–91.CrossRefPubMedPubMedCentral Karinja MN, Esterhuizen TM, Friedrich SO, Diacon AH. Sputum volume predicts sputum mycobacterial load during the first 2 weeks of antituberculosis treatment. J Clin Microbiol. 2015;53:1087–91.CrossRefPubMedPubMedCentral
12.
go back to reference Flores LL, Steingart KR, Dendukuri N, Schiller I, Minion J, Pai M, Ramsay A, Henry M, Laal S. Systematic review and meta-analysis of antigen detection tests for the diagnosis of tuberculosis. Clin Vaccine Immunol. 2011;18:1616–27.CrossRefPubMedPubMedCentral Flores LL, Steingart KR, Dendukuri N, Schiller I, Minion J, Pai M, Ramsay A, Henry M, Laal S. Systematic review and meta-analysis of antigen detection tests for the diagnosis of tuberculosis. Clin Vaccine Immunol. 2011;18:1616–27.CrossRefPubMedPubMedCentral
13.
go back to reference Peter JG, Zijenah LS, Chanda D, Clowes P, Lesosky M, Gina P, Mehta N, Calligaro G, Lombard CJ, Kadzirange G, Bandason T, Chansa A, Liusha N, Mangu C, Mtafya B, Msila H, Rachow A, Hoelscher M, Mwaba P, Theron G, Dheda K. Effect on mortality of point-of-care, urine-based lipoarabinomannan testing to guide tuberculosis treatment initiation in HIV-positive hospital inpatients: a pragmatic, parallel-group, multicountry, open-label, randomised controlled trial. Lancet. 2016;387(10024):1187–97.CrossRefPubMed Peter JG, Zijenah LS, Chanda D, Clowes P, Lesosky M, Gina P, Mehta N, Calligaro G, Lombard CJ, Kadzirange G, Bandason T, Chansa A, Liusha N, Mangu C, Mtafya B, Msila H, Rachow A, Hoelscher M, Mwaba P, Theron G, Dheda K. Effect on mortality of point-of-care, urine-based lipoarabinomannan testing to guide tuberculosis treatment initiation in HIV-positive hospital inpatients: a pragmatic, parallel-group, multicountry, open-label, randomised controlled trial. Lancet. 2016;387(10024):1187–97.CrossRefPubMed
14.
go back to reference Drain PK, Gounder L, Sahid F, Moosa M-YS. Rapid urine lam testing improves diagnosis of expectorated smear-negative pulmonary tuberculosis in an HIV-endemic region. Sci Rep. 2016;6:19992.CrossRefPubMedPubMedCentral Drain PK, Gounder L, Sahid F, Moosa M-YS. Rapid urine lam testing improves diagnosis of expectorated smear-negative pulmonary tuberculosis in an HIV-endemic region. Sci Rep. 2016;6:19992.CrossRefPubMedPubMedCentral
15.
go back to reference Kim SH, Lee NE, Lee JS, Shin JH, Lee JY, Ko J-H, Chang CL, Kim Y-S. Identification of mycobacterial antigens in human urine using immunoglobulin G isolated from sera of patients with active pulmonary tuberculosis. J Clin Microbiol. 2016;54(6):1631–7.CrossRefPubMedPubMedCentral Kim SH, Lee NE, Lee JS, Shin JH, Lee JY, Ko J-H, Chang CL, Kim Y-S. Identification of mycobacterial antigens in human urine using immunoglobulin G isolated from sera of patients with active pulmonary tuberculosis. J Clin Microbiol. 2016;54(6):1631–7.CrossRefPubMedPubMedCentral
16.
go back to reference Green C, Huggett JF, Talbot E, Mwaba P, Reither K, Zumla AI. Rapid diagnosis of tuberculosis through the detection of mycobacterial DNA in urine by nucleic acid amplification methods. Lancet Infect Dis. 2009;9:505–11.CrossRefPubMed Green C, Huggett JF, Talbot E, Mwaba P, Reither K, Zumla AI. Rapid diagnosis of tuberculosis through the detection of mycobacterial DNA in urine by nucleic acid amplification methods. Lancet Infect Dis. 2009;9:505–11.CrossRefPubMed
17.
go back to reference Dheda K, Davids V, Lenders L, Roberts T, Meldau R, Ling D, Brunet L, RvZ Smit, Peter J, Green C, Badri M, Sechi L, Sharma S, Hoelscher M, Dawson R, Whitelaw A, Blackburn J, Pai M, Zumla A. Clinical utility of a commercial LAM-ELISA assay for TB diagnosis in HIV-infected patients using urine and sputum samples. PLoS One. 2010;5:e9848.CrossRefPubMedPubMedCentral Dheda K, Davids V, Lenders L, Roberts T, Meldau R, Ling D, Brunet L, RvZ Smit, Peter J, Green C, Badri M, Sechi L, Sharma S, Hoelscher M, Dawson R, Whitelaw A, Blackburn J, Pai M, Zumla A. Clinical utility of a commercial LAM-ELISA assay for TB diagnosis in HIV-infected patients using urine and sputum samples. PLoS One. 2010;5:e9848.CrossRefPubMedPubMedCentral
18.
go back to reference Su Y-H, Wang M, Brenner DE, Ng A, Melkonyan H, Umansky S, Syngal S, Block TM. Human urine contains small, 150 to 250 nucleotide-sized, soluble DNA derived from the circulation and may be useful in the detection of colorectal cancer. J Mol Diagn. 2004;6:101–7.CrossRefPubMedPubMedCentral Su Y-H, Wang M, Brenner DE, Ng A, Melkonyan H, Umansky S, Syngal S, Block TM. Human urine contains small, 150 to 250 nucleotide-sized, soluble DNA derived from the circulation and may be useful in the detection of colorectal cancer. J Mol Diagn. 2004;6:101–7.CrossRefPubMedPubMedCentral
19.
go back to reference Bjerrum S, Kenu E, Lartey M, Newman MJ, Addo KK, Andersen AB, Johansen IS. Diagnostic accuracy of the rapid urine lipoarabinomannan test for pulmonary tuberculosis among HIV-infected adults in Ghana–findings from the DETECT HIV-TB study. BMC Infect Dis. 2015;15:407.CrossRefPubMedPubMedCentral Bjerrum S, Kenu E, Lartey M, Newman MJ, Addo KK, Andersen AB, Johansen IS. Diagnostic accuracy of the rapid urine lipoarabinomannan test for pulmonary tuberculosis among HIV-infected adults in Ghana–findings from the DETECT HIV-TB study. BMC Infect Dis. 2015;15:407.CrossRefPubMedPubMedCentral
20.
go back to reference Marangu D, Devine B, John-Stewart G. Diagnostic accuracy of nucleic acid amplification tests in urine for pulmonary tuberculosis: a meta-analysis. Int J Tuberc Lung Dis. 2015;19:1339–47.CrossRefPubMedPubMedCentral Marangu D, Devine B, John-Stewart G. Diagnostic accuracy of nucleic acid amplification tests in urine for pulmonary tuberculosis: a meta-analysis. Int J Tuberc Lung Dis. 2015;19:1339–47.CrossRefPubMedPubMedCentral
21.
go back to reference Kafwabulula M, Ahmed K, Nagatake T, Gotoh J, Mitarai S, Oizumi K, Zumla A. Evaluation of PCR-based methods for the diagnosis of tuberculosis by identification of mycobacterial DNA in urine samples. Int J Tuberc Lung Dis. 2002;6:732–7.PubMed Kafwabulula M, Ahmed K, Nagatake T, Gotoh J, Mitarai S, Oizumi K, Zumla A. Evaluation of PCR-based methods for the diagnosis of tuberculosis by identification of mycobacterial DNA in urine samples. Int J Tuberc Lung Dis. 2002;6:732–7.PubMed
22.
go back to reference Aceti A, Zanetti S, Mura MS, La Sechi, Turrini F, Saba F, Babudieri S, Mannu F, Fadda G. Identification of HIV patients with active pulmonary tuberculosis using urine based polymerase chain reaction assay. Thorax. 1999;54:145–6.CrossRefPubMedPubMedCentral Aceti A, Zanetti S, Mura MS, La Sechi, Turrini F, Saba F, Babudieri S, Mannu F, Fadda G. Identification of HIV patients with active pulmonary tuberculosis using urine based polymerase chain reaction assay. Thorax. 1999;54:145–6.CrossRefPubMedPubMedCentral
23.
go back to reference Peter JG, Theron G, Muchinga TE, Govender U, Dheda K. The diagnostic accuracy of urine-based Xpert MTB/RIF in HIV-infected hospitalized patients who are smear-negative or sputum scarce. PLoS One. 2012;7:e39966.CrossRefPubMedPubMedCentral Peter JG, Theron G, Muchinga TE, Govender U, Dheda K. The diagnostic accuracy of urine-based Xpert MTB/RIF in HIV-infected hospitalized patients who are smear-negative or sputum scarce. PLoS One. 2012;7:e39966.CrossRefPubMedPubMedCentral
24.
go back to reference Cannas A, Goletti D, Girardi E, Chiacchio T, Calvo L, Cuzzi G, Piacentini M, Melkonyan H, Umansky SR, Lauria FN, et al. Mycobacterium tuberculosis DNA detection in soluble fraction of urine from pulmonary tuberculosis patients. Int J Tuberc Lung Dis. 2008;12:146–51.PubMed Cannas A, Goletti D, Girardi E, Chiacchio T, Calvo L, Cuzzi G, Piacentini M, Melkonyan H, Umansky SR, Lauria FN, et al. Mycobacterium tuberculosis DNA detection in soluble fraction of urine from pulmonary tuberculosis patients. Int J Tuberc Lung Dis. 2008;12:146–51.PubMed
25.
go back to reference Gopinath K, Singh S. Urine as an adjunct specimen for the diagnosis of active pulmonary tuberculosis. Int J Infect Dis. 2009;13(3):374–9.CrossRefPubMed Gopinath K, Singh S. Urine as an adjunct specimen for the diagnosis of active pulmonary tuberculosis. Int J Infect Dis. 2009;13(3):374–9.CrossRefPubMed
26.
go back to reference Ralph AP, Ardian M, Wiguna A, Maguire GP, Becker NG, Drogumuller G, Wilks MJ, Waramori G, Tjitra E, Sandjaja Kenagalem E, Pontororing GJ, Anstey NM, Kelly PM. A simple, valid, numerical score for grading chest x-ray severity in adult smear-positive pulmonary tuberculosis. Thorax. 2010;65:863–9. doi:10.1136/thx.2010.136242.CrossRefPubMed Ralph AP, Ardian M, Wiguna A, Maguire GP, Becker NG, Drogumuller G, Wilks MJ, Waramori G, Tjitra E, Sandjaja Kenagalem E, Pontororing GJ, Anstey NM, Kelly PM. A simple, valid, numerical score for grading chest x-ray severity in adult smear-positive pulmonary tuberculosis. Thorax. 2010;65:863–9. doi:10.​1136/​thx.​2010.​136242.CrossRefPubMed
27.
go back to reference Newcombe RG. Two-sided confidence intervals for the single proportion: comparison of seven methods. Stat Med. 1998;17(8):857–72.CrossRefPubMed Newcombe RG. Two-sided confidence intervals for the single proportion: comparison of seven methods. Stat Med. 1998;17(8):857–72.CrossRefPubMed
28.
go back to reference Heyckendorf J, Olaru ID, Ruhwald M, Lange C. Getting personal perspectives on individualized treatment duration in multidrug-resistant and extensively drug-resistant tuberculosis. Am J Respir Crit Care Med. 2014;190:374–83.CrossRefPubMed Heyckendorf J, Olaru ID, Ruhwald M, Lange C. Getting personal perspectives on individualized treatment duration in multidrug-resistant and extensively drug-resistant tuberculosis. Am J Respir Crit Care Med. 2014;190:374–83.CrossRefPubMed
29.
go back to reference Olaru ID, von Groote-Bidlingmaier F, Heyckendorf J, Yew WW, Lange C, Chang KC. Novel drugs against tuberculosis: a clinician’s perspective. Eur Respir J. 2014;45:1119–31.CrossRefPubMed Olaru ID, von Groote-Bidlingmaier F, Heyckendorf J, Yew WW, Lange C, Chang KC. Novel drugs against tuberculosis: a clinician’s perspective. Eur Respir J. 2014;45:1119–31.CrossRefPubMed
Metadata
Title
Detection of transrenal DNA for the diagnosis of pulmonary tuberculosis and treatment monitoring
Authors
Ines Labugger
Jan Heyckendorf
Stefan Dees
Emilia Häussinger
Christian Herzmann
Thomas A. Kohl
Elvira Richter
Eric Rivera-Milla
Christoph Lange
Publication date
01-06-2017
Publisher
Springer Berlin Heidelberg
Published in
Infection / Issue 3/2017
Print ISSN: 0300-8126
Electronic ISSN: 1439-0973
DOI
https://doi.org/10.1007/s15010-016-0955-2

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