Development of a rapid method for determining the susceptibility of Mycobacterium tuberculosis to isoniazid using the Gen-Probe DNA Hybridization System

Kawa, Diane; Pennell, D R; Kubista, Louise N.; Schell, Ronald F.

doi:10.1128/aac.33.7.1000

Cited by 23 publications

(18 citation statements)

References 16 publications

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“…Several methods have been developed that decrease greatly the time required by the proportion method to obtain susceptibility test results [11][12][13][14][15][16][17][18][19][20]. These methods include a bioluminescence assay for detection of mycobacterial ATP [11], the Gen-Probe DNA hybridisation system [12], the luciferase reporter gene assay [14], high-performance liquid chromatography mycolic acid analysis [15], the Etest method [16] and a colorimetric method [17]. However, to date, only the BACTEC method has been recognised as a valid alternative [7,8].…”

Section: Discussionmentioning

confidence: 99%

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Susceptibility testing of Mycobacterium tuberculosis: comparison of the BACTEC TB-460 method and flow cytometric assay with the proportion method

DeCoster

Vena

Callister

et al. 2005

Clinical Microbiology and Infection

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Tuberculosis is a leading cause of morbidity and mortality worldwide. Susceptibility testing of the causative agent, Mycobacterium tuberculosis, is critical for control of the disease. This study compared the flow cytometric susceptibility assay with the proportion method and the BACTEC TB-460 system. There was agreement between the flow cytometric and proportion methods for 73 (94%) of 78 isoniazid tests, and complete agreement for 26 ethambutol and rifampicin tests. In contrast, the proportion and BACTEC methods failed to agree for 22%, 15% and 8% of isoniazid, ethambutol and rifampicin tests, respectively. These findings indicated that susceptibility testing by the flow cytometric assay is accurate, with results available within 24 h of initiation of the testing procedure.

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Section: Discussionmentioning

confidence: 99%

“…Even the method used most frequently, BACTEC-460, requires incubation for 4-12 days [4][5][6][7][8]. Therefore, newer methods are being developed constantly to facilitate more rapid availability of susceptibility results [9][10][11][12][13][14][15][16][17][18].…”

Section: Introductionmentioning

confidence: 99%

Susceptibility testing of Mycobacterium tuberculosis: comparison of the BACTEC TB-460 method and flow cytometric assay with the proportion method

DeCoster

Vena

Callister

et al. 2005

Clinical Microbiology and Infection

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“…One such approach has been the application of the Gen-Probe MTD amplification assay to the BACTEC system (Kawa et al 1989;Miyamoto et al 1996;Martin-Casabona et al 1997). Molecular methods that provide a direct measurement of bacterial metabolism can potentially circumvent this problem.…”

Section: Metabolic Assays For Assessing Viability In the Presence Of mentioning

confidence: 99%

Molecular Diagnostics

Eisenach

1999

Mycobacteria

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“…Das System BACTEC 460TB nutzt allerdings ein radiometrisches Meßprinzip, was einer breiten Anwendung hinderlich ist. Es gibt deshalb Bemühungen, andere Nachweissysteme für die Empfindlichkeitstestung von Mykobakterien zu etablieren [15][16][17][18][19][20][21][22][23][24][25]. In der vorliegenden Arbeit haben wir die Möglichkeit geprüft,' mit dem für die Mykobakteriendiagnostik entwickelten Kultursystem MB/BacT (Organon Teknika Corp.) die Empfindlichkeit von Mykobakterien gegenüber verschiedenen Antituberkulotika zu bestimmen.…”

Section: Er Einsatz Von Flüssigkulturen Im Mykobakterien-unclassified

Untersuchungen zur Möglichkeit der Empfindlichkeitstestung von Mykobakterien mit dem MB/BacT-Kultursystem

Beer¹,

Küchler²,

Rodloff³

1997

LaboratoriumsMedizin

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Zusammenfassung: Es wurde eine Methode zur Empfindlichkeitstestung von Mykobakterien in Flüssigkulturmedien ohne Verwendung radioaktiver Isotope entwickelt: Kommerziell erhältliche MB/BacT-Kulturflaschen wurden mit geeigneten Konzentrationen von Antituberkulotika versetzt, anschließend mit einer Mykobakterien-Testsuspension (MacFarland Nr. 1) beimpft und im MB/BacT-System bebrütet. Bei der Untersuchung von Pyrazinamid (PZA) wurde außerdem der pH-Wert in den Kulturflaschen mit 2? 4 auf 5,5 abgesenkt. Eine Antituberkulotika-freie Kulturflasche, die mit der 1:100 verdünnten Testsuspension beimpft wurde, diente als Kontrolle (1%-Kontrolle). Wenn vom MB/BacT-System in dieser Kontrollflasche Wachstum erkannt wurde (nach ca. 7 Tagen), wurde die Untersuchung beendet und der bis zu diesem Zeitpunkt in den Antituberkulotika-haltigen Kulturflaschen festgestellte Wachstumsstatus bewertet. Für ausgewählte M. tuberciilosis-Stämme wurden die MHK-Werte folgender Antituberkulotika bestimmt: Ethambutol (EMB), Isoniazid (INH), Rifampicin (RMP), Streptomycin (SM), Dihydrostreptomycin, Protionamid und PZA. Die Testung von 16 M. tuberculosis-Stämmen und 3 Stämmen atypischer Mykobakterien gegenüber EMB (2,5 mg/1), INH (1,0 mg/1), RMP (2,0 mg/1), SM (2,0 mg/1) und PZA (50 mg/1) ergab die gleichen Empfindlichkeitsbewertungen, wie sie mit Referenzmethoden (Verdünnungstest mit Löwenstein-Jensen-Medium, BACTEC 460TB) erhalten wurden. Schlüsselwörter: Mikrobielle Empfindlichkeitstestung/Methodik; Mycobacterium tuberculosis/Medikamenteneffekte; Antituberkulotika. 1 Teile dieser Arbeit wurden auf der 48. Jahrestagung der DeutschenSummary: A method for susceptibility testing of mycobacteria using isotope-free fluid culture media was developed. Commercially available MB/BacT process bottles supplemented with antituberculotics in adequate concentrations were inoculated with a mycobacteria test Suspension (MacFarland No.l) and loaded into the System. In case of testing pyrazinamide (PZA), the medium in the culture bottles was acidified to pH 5.5 by 2? 4 . bottle without antituberculotics inoculated with a 1:100 dilution of the test suspension served äs a control (1% control). At the time the MB/BacT recognized growth in the control bottle, the test was terminated (after approximately 7 days) and the growth Status of the bottles containing antituberculotics was determined. The minimum inhibitory concentrations of ethambutol (EMB), isoniazid (INH), rifampicin (RMP), streptomycin (SM), dihydrostreptomycin, prothionamid, and PZA were determined for selected strains of M. tuberculosis. The susceptibility testing of 16 strains of M. tuberculosis and 3 mycobacteria other than tubercle bacilli to EMB (2.5 mg/1), INH (1.0 mg/1), RMP (2.0 mg/1), SM (2.0 mg/1), and PZA (50 mg/1) exhibited the same susceptibility patteras äs Standard methods (dilution method with Löwenstein-Jensen medium, BACTEC 460 TB).

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Development of a rapid method for determining the susceptibility of Mycobacterium tuberculosis to isoniazid using the Gen-Probe DNA Hybridization System

Abstract: tuberculosis to other antimycobacterial agents and its correlation with clinically significant levels of resistance.

Cited by 23 publications

References 16 publications

Susceptibility testing of Mycobacterium tuberculosis: comparison of the BACTEC TB-460 method and flow cytometric assay with the proportion method

Susceptibility testing of Mycobacterium tuberculosis: comparison of the BACTEC TB-460 method and flow cytometric assay with the proportion method

Molecular Diagnostics

Untersuchungen zur Möglichkeit der Empfindlichkeitstestung von Mykobakterien mit dem MB/BacT-Kultursystem

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