Evaluation Of The Performance Of The Nitrate Reductase And Resazurin Titre Assay For The Detection of Mycobacterium Tuberculosis Complex From Sputum In A High Tb and Hiv Setting (NRA RETA)

EVALUATION OF THE PERFORMANCE OF THE NITRATE REDUCTASE AND RESAZURIN TITRE ASSAY FOR THE DETECTION OF MYCOBACTERIUM TUBERCULOSIS COMPLEX FROM SPUTUM IN A HIGH TB AND HIV SETTING

The Principle objective of this study is To evaluate the performance of NRA, NRA-p and REMA-p for the detection of M. tuberculosis complex from sputum samples from adult pulmonary TB suspects in a high TB and HIV prevalence setting, using LJ and MGIT culture as gold standard.

The Secondary objectives are:

• To measure the performance of each assay (NRA, NRA-p, REMA-p) in sputum smear-negative patients
• To describe the results of the colorimetric methods in HIV-positive and HIV-negative patients
• To assess the time to detection of both NRA/NRA-p, REMA-p methods.
• To evaluate the feasibility of the NRA, NRA-p, REMA-p methods.
• To determine the rate of contamination of the NRA, NRA-p and REMA-p assays.
• To evaluate the proportion and the clinical relevance of NTM among TB suspects in a high TB and HIV prevalence setting.
• To provide capacity building for TB diagnosis in Mbarara.

Study Overview

• Status: Completed
• Conditions: HIV Infections; HIV; Tuberculosis; AIDS

Detailed Description

Tuberculosis (TB) remains a leading cause of death in developing countries and its burden has been exacerbated by the concurrent HIV epidemic. Despite the advances in medicine, TB diagnosis still remains a challenge, especially in developing countries where diagnosis relies mostly on the detection of Mycobacterium tuberculosis complex (MTBC) by smear microscopy and/or culture. Smear microscopy is rapid, simple and not expensive but it lacks sensitivity. Culture on solid medium, which is performed in some well equipped laboratories, is more sensitive than microscopy but takes up to 8 weeks to obtain the result.

Colorimetric methods have been used for the rapid detection of drug sensitivity in M. tuberculosis either from isolates or directly from sputum. These methods rely on the detection of live bacteria through either enzymatic activity (nitrate reduction) or their ability to reduce an oxidation-reduction indicator, either in solid or liquid medium. They are fast, simple, and offer a good potential that should be evaluated for the diagnosis of TB.

The objective of this study is to evaluate the performance of colorimetric assays for the detection of M. tuberculosis complex from sputum samples from adult pulmonary TB suspects in a high TB and HIV prevalence setting, using Löwenstein Jensen (LJ) and Mycobacterium growth indicator tube (MGIT) culture as gold standard. The colorimetric methods evaluated here will be the solid medium-based nitrate reductase assay as described (NRA) or with an additional step using para Nitrobenzoic (PNB) acid for differentiation of MTBC and NTM (NRA-p), and the modified resazurin microplate assay, also using PNB for differentiation of MTBC and NTM (REMA-p).

. If any of the colorimetric assays is found to be accurate, significantly faster than conventional culture methods, and easy to perform, then it could be implemented in a tuberculosis culture laboratory. By reducing the time to detection compared to conventional culture and the costs compared to recent commercial methods, these assays offer a good alternative to conventional methods and might help to improve TB diagnosis in developing countries.

• Study Type: Observational
• Enrollment (Anticipated): 600

Contacts and Locations

Study Locations

• Uganda
  • Mbarara, Uganda, 1956
    • Epicentre

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 14 years and older (ADULT, OLDER_ADULT, CHILD)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Probability Sample

Study Population

All patients presenting at the outpatient department of the MRRH will be eligible if they meet the inclusion criteria

Description

Inclusion Criteria:

• Pulmonary TB suspects as defined by a cough for more than 2 weeks
• Age 15 years and above
• Written informed consent signed

Exclusion Criteria:

• Producing pure blood sputum or clear saliva
• Producing less than 1 mL of sputum
• Receiving anti TB treatment (isoniazid, rifampicin, streptomycin, pyrazinamide, ethambutol) or quinolone for more than 1 week in the month before inclusion.

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Time Frame
To evaluate the performance of NRA, NRA-p and REMA-p for the detection of M. tuberculosis complex from sputum samples from adult pulmonary TB suspects in a high TB and HIV prevalence setting, using LJ and MGIT culture as gold standard	7 Month

Secondary Outcome Measures

Outcome Measure	Time Frame
-To measure the performance of each assay (NRA, NRA-p, REMA-p) in sputum smear-negative patients -To describe the results of the colorimetric methods in HIV-positive and HIV-negative patients	7 Month

Collaborators and Investigators

• Sponsor: Epicentre
• Collaborators: Mbarara University of Science and Technology; Medecins Sans Frontieres, Netherlands

Study record dates

Study Major Dates

• Study Start: September 1, 2011
• Primary Completion (ACTUAL): September 1, 2012
• Study Completion (ACTUAL): December 1, 2012

Study Registration Dates

• First Submitted: January 19, 2010
• First Submitted That Met QC Criteria: January 20, 2010
• First Posted (ESTIMATE): January 21, 2010

Study Record Updates

• Last Update Posted (ESTIMATE): February 20, 2013
• Last Update Submitted That Met QC Criteria: February 16, 2013
• Last Verified: February 1, 2013

More Information

Terms related to this study

• Keywords: Mycobacterium tuberculosis; Nitrate Reductase Assay; NRA; REMA; RETA; Rezazurine
• Additional Relevant MeSH Terms: Infections; Bacterial Infections; Bacterial Infections and Mycoses; Gram-Positive Bacterial Infections; Actinomycetales Infections; Mycobacterium Infections; Tuberculosis
• Other Study ID Numbers: 821183