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Evaluation of Dual-Color Fluorescence In Situ Hybridization With Peptide Nucleic Acid Probes for the Detection of Mycobacterium tuberculosis and Non-Tuberculous Mycobacteria in Clinical Specimens
Annals of Laboratory Medicine ; : 500-505, 2015.
Article Dans Anglais | WPRIM | ID: wpr-110966
ABSTRACT

BACKGROUND:

Peptide nucleic acid (PNA) probes are artificial DNA analogues with a hydrophobic nature that can penetrate the mycobacterial cell wall. We evaluated a FISH method for simultaneous detection and identification of Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM) in clinical respiratory specimens using differentially labeled PNA probes.

METHODS:

PNA probes targeting the mycobacterial 16S ribosomal RNA were synthesized. The cross-reactivity of MTB- and NTM-specific probes was examined with reference strains and 10 other frequently isolated bacterial species. A total of 140 sputum specimens were analyzed, comprising 100 MTB-positive specimens, 21 NTM-positive specimens, and 19 MTB/NTM-negative specimens; all of them were previously confirmed by PCR and culture. The PNA FISH test results were graded by using the United States Centers for Disease Control and Prevention-recommended scale and compared with the results from the fluorochrome acid-fast bacterial stain.

RESULTS:

The MTB- and NTM-specific PNA probes showed no cross-reactivity with other tested bacterial species. The test results demonstrated 82.9% agreement with the culture results with diagnostic sensitivity of 80.2% and diagnostic specificity of 100.0% (kappa=0.52, 95% confidence interval 0.370-0.676).

CONCLUSIONS:

Dual-color PNA FISH showed high specificity for detecting and identifying mycobacteria in clinical specimens. However, because of its relatively low sensitivity, this method could be more applicable to culture confirmation. In application to direct specimens, the possibility of false-negative results needs to be considered.
Sujets)

Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) Sujet Principal: Expectoration / ADN / ARN ribosomique 16S / Sondes d'acide nucléique / Paroi cellulaire / Réaction de polymérisation en chaîne / Hybridation in situ / Acides nucléiques peptidiques / Fluorescence / Mycobacterium tuberculosis Type d'étude: Etude diagnostique langue: Anglais Texte intégral: Annals of Laboratory Medicine Année: 2015 Type: Article

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Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) Sujet Principal: Expectoration / ADN / ARN ribosomique 16S / Sondes d'acide nucléique / Paroi cellulaire / Réaction de polymérisation en chaîne / Hybridation in situ / Acides nucléiques peptidiques / Fluorescence / Mycobacterium tuberculosis Type d'étude: Etude diagnostique langue: Anglais Texte intégral: Annals of Laboratory Medicine Année: 2015 Type: Article