Evaluation of two different cell lysis methods for releasing mycobacterial nucleic acids in the INNO-LiPA mycobacteria test.
The INNO-LiPA Mycobacteria Test (Innogenetics, N.V., Belgium) is a PCR-based reverse hybridization assay for the simultaneous identification of several mycobacterial species. We evaluated two simplified lysis methods for mycobacterial DNA release for application in the INNO-LiPA Mycobacteria Test. The two methods were based on either (i) heat treatment or (ii) sonication. Both methods were performed directly on 45 positive liquid cultures (MB-BacT, BioMérieux, Marcy l'Etoile, France) containing 17 different mycobacterial species. These two simple lysis procedures demonstrated similar effectiveness (100%) to that recommended by the manufacturer. They also significantly shortened the time required for mycobacterial DNA release.
Padilla
E
E
Servicio de Microbiología, Departamento de Genética y Microbiología, Hospital Universitario Germans Trias i Pujol, Universidad Autónoma de Barcelona, Barcelona, Spain. epadilla.@ns.hugtip.scs.es
González
V
V
Manterola
J M
JM
Lonca
J
J
Pérez
A
A
Matas
L
L
Quesada
M D
MD
Ausina
V
V
D015373:Bacterial Typing Techniques / Q000379:methods*; D002468:Cell Physiological Phenomena; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D009161:Mycobacterium / Q000145:classification* / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques; D009696:Nucleic Acids / Q000032:analysis / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
D003470:Culture Media; D004269:DNA, Bacterial; D009696:Nucleic Acids; D011933:Reagent Kits, Diagnostic