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Year : 2018  |  Volume : 7  |  Issue : 3  |  Page : 217-221

Mycobacterial inactivation protein extraction protocol for matrix-assisted laser desorption ionization time-of-flight characterization of clinical isolates

1 Microbiology and Infectious Diseases Directorate, SA Pathology, Royal Adelaide Hospital, Adelaide South, Australia
2 Microbiology and Infectious Diseases Directorate, SA Pathology, Royal Adelaide Hospital; Department of Molecular and Biomedical Science, University of Adelaide, Adelaide South, Australia

Correspondence Address:
Mr. Manuel Pastor Forero Morales
1 North Terrace, Royal Adelaide Hospital, Level 3, SA Pathology, Microbiology and Infectious Diseases Laboratory, Adelaide 5000, South Australia
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/ijmy.ijmy_104_18

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Background: Rapid identification of mycobacteria has been made possible with matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) in recent years. Working with high concentrations of mycobacteria in a PC-3 containment facility makes MALDI-TOF cumbersome and costly. Therefore removing the inactivated isolate's protein extract from the PC-3 facility is needed for efficient identification in a routine PC-2 laboratory. Methods: This work describes a novel chemical and mechanical disruption protein extraction method, which provides reliable MALDI-TOF results from solid and liquid media, while ensuring laboratory safety. Results: When compared to sequencing results, 93.9% of the clinical isolates were identified in LJ media and 89% of the clinical isolates were identified in MGIT media. Conclusion: The MIPE protocol produces a high quality protein extract with improved isolate identification without compromising result turn-around-times or laboratory safety.

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