ORIGINAL PAPER
Drug susceptibility testing of first-line anti-tuberculosis drugs: clinical evaluation of the differences between laboratories
 
More details
Hide details
1
Pulmonary Department, Aristotle University of Thessaloniki, “G. Papanikolaou” Hospital, Thessaloniki, Greece
 
2
Greek National Reference Laboratory for Mycobacteria, General Hospital for Chest Diseases “SOTIRIA”, Athens, Greece
 
3
Laboratory for Tuberculosis of Northern Greece, “G. Papanikolaou” Hospital, Thessaloniki, Greece
 
 
Corresponding author
Ioannis Kioumis   

Pulmonary Department, Aristotle University of Thessaloniki, “G. Papanikolaou” Hospital, 57010 Exohi, Thessaloniki, Greece
 
 
Pneumon 2017;30(1):24-31
 
KEYWORDS
ABSTRACT
Background:
Tuberculosis (TB) remains a significant public health issue, partly due to drug resistance development. The rapid and reliable diagnosis is essential for TB control. The purpose of the present study was to detect and present the differences of molecular and phenotypic drug susceptibility testing (DST) of Μ. tuberculosis strains, isolated from patients hospitalized in the Pulmonary Department of the Aristotle University of Thessaloniki, as they were recorded by the Greek National Reference Laboratory for Mycobacteria (NRLM) (General Hospital for Chest Diseases “SOTIRIA”, Athens, Greece) and the Laboratory for Tuberculosis (LT) of Northern Greece, G.H. “G. PAPANIKOLAOU”.

Methods:
Twenty-one ΤΒ patients were included. Culture of the Mycobacterium strain was conducted using Lowenstein-Jensen medium and phenotypic antibiograms were obtained from NRLM and LT. Molecular testing was conducted by NRLM. Isoniazid and rifampicin were tested.

Results:
Eleven isolates yielded discrepant DST results. For isoniazid, 6 cases were found to be molecularly and phenotypically susceptible by NRLM while resistant by LT. In 3 cases, resistance was attributed to a problematic reagent. For rifampicin, four molecularly susceptible strains demonstrated phenotypic susceptibility at NRLM but resistance at LT. In one case, resistance was taken into account for treatment interventions. Furthermore, one strain demonstrated molecular and phenotypic resistance to rifampicin at NRLM, but susceptibility at LT. The strain was molecularly and phenotypically resistant to isoniazid and the patient was considered as a case of multidrug-resistant TB.

Conclusions:
Discordance between DST requires full consideration of the clinical presentation and collaboration with the laboratory and TB experts.

REFERENCES (34)
1.
European Centre for Disease Prevention and Control. Annual Epidemiological Report 2016 – Tuberculosis. Stockholm: ECDC; 2016. Available from: http://ecdc.europa.eu/en/healt....
 
2.
European Centre for Disease Prevention and Control, WHO Regional Office for Europe. Tuberculosis surveillance and monitoring in Europe 2016. Stockholm: ECDC; 2016.
 
3.
Κέντρο Ελέγχου & Πρόληψης Νοσημάτων (ΚΕΕΛΠΝΟ), Επιδημιολογικά δεδομένα φυματίωσης στην Ελλάδα, 2004-2010.
 
4.
Κέντρο Ελέγχου & Πρόληψης Νοσημάτων (ΚΕΕΛΠΝΟ), Ενημερωτικό Δελτίο, Νεώτερα επιδημιολογικά δεδομένα για τη φυματίωση στην Ελλάδα. http://www2.keelpno.gr/blog/?p... .
 
5.
Lytras T, Spala G, Bonovas S, Panagiotopoulos T. Evaluation of Tuberculosis Underreporting in Greece through Comparison with Anti-Tuberculosis Drug Consumption. PLoS One 2012; 7:7-12. doi:10.1371/journal.pone.0050033.
 
6.
Ibarz-Pavón AB, Papaventsis D, Kalkouni R, et al. Pilot study of the completeness of notification of adult tuberculosis in Athens, Greece. Int J Tuberc Lung Dis 2016; 20:920-5. doi:10.5588/ijtld.15.0907.
 
7.
World Health Organization, Global Tuberculosis Report 2016 WHO/HTM/TB/2016.13, Geneva, Switzerland: WHO, 2016.
 
8.
Ahmad S, Mokaddas E, Al-Mutairi N, Eldeen HS, Mohammadi S. Discordance across Phenotypic and Molecular Methods for Drug Susceptibility Testing of Drug-Resistant Mycobacterium tuberculosis Isolates in a Low TB Incidence Country. PLoS One 2016;11:e0153563. doi:10.1371/journal.pone.0153563.
 
9.
Gandhi NR, Andrews JR, Brust JCM, et al. Risk factors for mortality among MDR- and XDR-TB patients in a high HIV prevalence setting.
 
10.
Lonnroth K, Migliori GB, Abubakar I, et al. Towards tuberculosis elimination: an action framework for low-incidence countries. Eur Respir J 2015; 45:928–52. doi: 10.1183/09031936.00214014.
 
11.
Drobniewski F, Nikolayevskyy V, Balabanova Y, Bang D, Papaventsis D. Diagnosis of tuberculosis and drug resistance: what can new tools bring us? Int J Tuberc Lung Dis 2012; 16:860–70. doi: 10.5588/ijtld.12.0180.
 
12.
ISO 15189:2012. Medical laboratories—Requirements for quality and competence. International Standard Organization 2012.
 
13.
Fattorini L, Iona E, Cirillo D, et al. External quality control of Mycobacterium tuberculosis drug susceptibility testing: results of two rounds in endemic countries. Int J Tuberc Lung Dis 2008;12:214-7.
 
14.
Nikolayevskyy V, Hillemann D, Richter E, et al. External Quality Assessment for Tuberculosis Diagnosis and Drug Resistance in the European Union: A Five Year Multicentre Implementation Study. PLoS One 2016; 11:e0152926. doi:10.1371/journal. pone.0152926.
 
15.
Hillemann D, Hoffner S, Cirillo D, Drobniewski F, Richter E, Rüsch-Gerdes S. First Evaluation after Implementation of a Quality Control System for the Second Line Drug Susceptibility Testing of Mycobacterium tuberculosis Joint Efforts in Low and High Incidence Countries. PLoS One 2013; 8:1-7. doi:10.1371/ journal.pone.0076765.
 
16.
Laszlo A, Rahman M, Espinal M, Raviglione M. Quality assurance programme for drug susceptibility testing of Mycobacterium tuberculosis in the WHO/IUATLD Supranational Reference Laboratory Network: five rounds of proficiency testing, 1994- 1998. IntJTubercLungDis 2002;6:748-56.
 
17.
European Centre for Disease Prevention and Control. ERLN-TB expert opinion on the use of the rapid molecular assays for the diagnosis of tuberculosis and detection of drug-resistance. Stockholm: ECDC; 2013.
 
18.
Κατευθυντήριες οδηγίες για τη θεραπεία της φυματίωσης στους ενήλικες. Πνεύμων 2015, 28:268.
 
19.
Steingart KR, Sohn H, Schiller I, et al. Xpert® MTB/RIF assay for pulmonary tuberculosis and rifampicin resistance in adults. Cochrane databaseSystRev. 2013;1:CD009593. doi:10.1002/14651858.CD009593.pub2.
 
20.
Barnard M, Albert H, Coetzee G, O’Brien R, ME Bosman. Rapid molecular screening for multidrug-resistant tuberculosis in a high- volume public health laboratory in South Africa. Am J Respir Crit Care Med 2008; 177:787–92.
 
21.
Yakrus MA, Driscoll J, Lentz AJ, et al. Concordance between molecular and phenotypic testing of mycobacterium tuberculosis complex isolates for resistance to rifampin and isoniazid in the United States. J Clin Microbiol 2014;52:1932-7. doi:10.1128/ JCM.00417-14.
 
22.
Banu S, Rahman SMM, Khan MSR, et al. Discordance across several methods for drug susceptibility testing of drug-resistant Mycobacterium tuberculosis isolates in a single laboratory. J Clin Microbiol 2014; 52:156-63. doi:10.1128/JCM.02378-13.
 
23.
Van Deun A, Barrera L, Bastian I, et al. Mycobacterium tuberculosis strains with highly discordant rifampin susceptibility test results. J Clin Microbiol 2009; 47:3501-6. doi:10.1128/ JCM.01209-09.
 
24.
Rigouts L, Gumusboga M, de Rijk WB, et al. Rifampin Resistance Missed in Automated Liquid Culture System for Mycobacterium tuberculosis Isolates with Specific rpoB Mutations. Journal of Clinical Microbiology 2013; 51:2641-5. doi:10.1128/JCM.02741- 12.
 
25.
World Health Organization. Updated interim critical concentrations for first-line and second-line DST (as of May 2012). 2012; http://www.stoptb.org/wg/gli/a....
 
26.
Dominguez et al. Clinical implications of molecular drug resistance testing for Mycobacterium tuberculosis: a TBNET/ RESIST-TB consensus statement. Int J Tuberc Lung Dis 2016; 20:24-42. doi: 10.5588/ijtld.15.0221.
 
27.
van Rie A, et al. 2005. Reinfection and mixed infection cause changing Mycobacterium tuberculosis drug-resistance patterns. Am J Respir Crit Care Med 172:636–42.
 
28.
Cohen T, van Helden PD, Wilson D, et al. Mixed-strain Mycobacterium tuberculosis infections and the implications for tuberculosis treatment and control. Clin Microbiol Rev 2012; 25:708-19. doi:10.1128/CMR.00021-12.
 
29.
van Deun A, Aung KJM, Bola V, et al. Rifampin drug resistance tests for tuberculosis: challenging the gold standard. J Clin Microbiol 2013; 51:2633-40. doi:10.1128/JCM.00553-13.
 
30.
van Ingen J, Aarnoutse R, de Vries G, Boeree MJ, van Soolingen D. Low-level rifampicin-resistant Mycobacterium tuberculosis strains raise a new therapeutic challenge. Int J Tuberc Lung Dis 2011;15:990-92. doi:10.5588/ijtld.10.0127.
 
31.
Williamson DA, Roberts SA, Bower JE, et al. Clinical failures associated with rpoB mutations in phenotypically occult multidrug-resistant Mycobacterium tuberculosis. Int J Tuberc Lung Dis 2012;16:216-20. doi:10.5588/ijtld.11.0178.
 
32.
Ioerger TR, Koo S, No E-G, et al. Genome analysis of multi- and extensively-drug-resistant tuberculosis from KwaZulu-Natal, South Africa. PLoS One 2009; 4:e7778. doi:10.1371/journal. pone.0007778.
 
33.
Τρυφινοπούλου Κ, Παπαβέντσης Δ, Ιωαννίδης Π, Βογιατζάκης ΕΔ. Καταγραφή μικροβιολογικών εργαστηρίων νοσηλευτικών ιδρυμάτων που εμπλέκονται στην εργαστηριακή διάγνωση της φυματίωσης. Περιγραφή εύρους διαγνωστικών εξετάσεων και δεικτών διασφάλισης ποιότητας και βιοασφάλειας. Μικροβιολογικό Εργαστήριο & Εθνικό Κέντρο Αναφοράς Μυκοβακτηριδίων, Αθήνα, 2017.
 
34.
Availability of an Assay for Detecting Mycobacterium tuberculosis, Including Rifampin-Resistant Strains, and Considerations for Its Use — United States, 2013, n.d. https://www.cdc.gov/mmwr/previ....
 
eISSN:1791-4914
ISSN:1105-848X
Journals System - logo
Scroll to top