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Rapid identification of mycobacterial species with test INNO-LiPA mycobacteria v2 isolated from sputum samples obtained from HIV-negative patients

Abstract number: p1107

Paluch-Oles  J., Koziol-Montewka  M., Dzbrowska  M., Montewka   M., Bogut  A.

The nontuberculous mycobacteria (NTM) are a group of bacteria that can cause pulmonary and systemic diseases. An increased risk of NTB infections is more frequent in patients with acquired immunodeficiency syndrome (AIDS) and HIV–negative patients suffering from immunosuppression due to previous pulmonary disease, old age and alcoholism. For the rapid detection and identification of mycobacteria new strategies have been developed. These novel methods such as HPLC, the BACTEC NAP test and commercial DNA probes are recommended for identification of NTM species. Recently, a new molecular test based on the PCR technique and reverse hybridization procedure has been introduced for simultaneous identification of different mycobacterial species.

Objectives: 

The aims of the study were to perform the DNA line probe assay – INNO-LiPA Mycobacteria v2 for identification of mycobacterial species obtained from sputum samples collected form HIV-negative patients and to evaluate the risk factors of pulmonary NTM infections.

Methods: 

We tested 15 NTM strains from patients with pulmonary tuberculosis symptoms, isolated in our centre within the last 2 years. All the strains were processed from subculture on LJ medium, confirmed as NTM using the niacin test and morphology assessment of the organisms. The LiPA assay based on the PCR and reverse hybridization procedure amplificated the 16S–23S ribosomal rRNA spacer region. Subsequently, the biotinylated PCR products were hybridized with specific 22 parallel DNA probes and 2 control lines. The clear pattern of lines allows simultaneous identification of 16 different mycobacterial species.

Results: 

Among 15 NTM isolates the LiPA revealed 6 isolates identified as M. kansasii group I, 4 isolates as M. xenopi, and 1 showed a MAIS pattern which also produced a positive result for specific M. intracellulare probe. These 3 species were identified in 11 alcoholic patients. In 4 patients older than 60 years 2 strains as Mycobacterium spp. and 2 as M. avium-M. intracellulare-M. scrofulaceum (MAIS) were identified.

Conclusion: 

This study demonstrated usefulness of the INNO-LiPA Mycobacteria v2 test for the rapid identification of mycobacterial species and revealed M. kansasii, M. xenopi and M. avium-complex species considered the most frequent mycobacteria causing lung disease in immunocompromised alcoholic and old patients.

Session Details

Date: 01/08/2007
Time: 00:00-00:00
Session name: XXIst ISTH Congress
Subject:
Location: Oxford, UK
Presentation type:
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