Jeong Su Park1, Soon Hee Choi2, Sang Mee Hwang1, Yun Ji Hong1, Taek Soo Kim2, Kyoung Un Park3, Junghan Song1, Eui-Chong Kim4. 1. Department of Laboratory Medicine, Bundang Seoul National University Hospital, Gyeonggi-do, Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea. 2. Department of Laboratory Medicine, Bundang Seoul National University Hospital, Gyeonggi-do, Korea. 3. Department of Laboratory Medicine, Bundang Seoul National University Hospital, Gyeonggi-do, Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea. Electronic address: m91w95pf@snu.ac.kr. 4. Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.
Abstract
BACKGROUND: Protein extraction step is particularly important for identification of mycobacterial isolates by MALDI-TOF mass spectrometry (MS) because of its thick and solid cell wall. This study compared the performance of MALDI-TOF MS for identification of mycobacterial clinical isolates cultured in liquid media between heating-based protocol and non-heating protocol. METHODS: Clinical mycobacterial isolates cultured in liquid media were prospectively analyzed. Reference identification was real-time PCR and restriction fragment length polymorphism. The specimens prepared by heating protocol and non-heating protocol were tested using MALDI Biotyper (Bruker Daltonics, Bremen, Germany) and Vitek MS (bioMérieux, Marcy l'Etoile, France), respectively. RESULTS: Among the 206 clinical specimens prepared by heating method, identification rates were 90.3% and 60.7% in MALDI Biotyper and Vitek MS, respectively. Identification accuracy of MALDI Biotyper and Vitek MS was 100% for the isolates of Mycobacterium tuberculosis complex (MTBC), Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus and Mycobacterium fortuitum. Among the 121 clinical specimens prepared by non-heating method, identification rate for MALDI Biotyper and Vitek MS were 61.2% and 69.4%, respectively. Identification accuracy of MALDI Biotyper/Vitek MS were 92.9%/94.1% for MTBC, 92.9%/100% for M. avium, 90%/100% for M. intracellulare, 100%/100% for M. abscessus and 100%/100% for M. fortuitum. CONCLUSIONS: The performance of MALDI-TOF MS for identification of mycobacterial clinical isolates is affected by protein extraction protocol. For best performance, protein extraction protocol should be chosen considering the MALDI-TOF MS system. In the present study, heating protocol with MALDI Biotyper system showed reliable identification results for mycobacterial clinical isolates.
BACKGROUND: Protein extraction step is particularly important for identification of mycobacterial isolates by MALDI-TOF mass spectrometry (MS) because of its thick and solid cell wall. This study compared the performance of MALDI-TOF MS for identification of mycobacterial clinical isolates cultured in liquid media between heating-based protocol and non-heating protocol. METHODS: Clinical mycobacterial isolates cultured in liquid media were prospectively analyzed. Reference identification was real-time PCR and restriction fragment length polymorphism. The specimens prepared by heating protocol and non-heating protocol were tested using MALDI Biotyper (Bruker Daltonics, Bremen, Germany) and Vitek MS (bioMérieux, Marcy l'Etoile, France), respectively. RESULTS: Among the 206 clinical specimens prepared by heating method, identification rates were 90.3% and 60.7% in MALDI Biotyper and Vitek MS, respectively. Identification accuracy of MALDI Biotyper and Vitek MS was 100% for the isolates of Mycobacterium tuberculosis complex (MTBC), Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus and Mycobacterium fortuitum. Among the 121 clinical specimens prepared by non-heating method, identification rate for MALDI Biotyper and Vitek MS were 61.2% and 69.4%, respectively. Identification accuracy of MALDI Biotyper/Vitek MS were 92.9%/94.1% for MTBC, 92.9%/100% for M. avium, 90%/100% for M. intracellulare, 100%/100% for M. abscessus and 100%/100% for M. fortuitum. CONCLUSIONS: The performance of MALDI-TOF MS for identification of mycobacterial clinical isolates is affected by protein extraction protocol. For best performance, protein extraction protocol should be chosen considering the MALDI-TOF MS system. In the present study, heating protocol with MALDI Biotyper system showed reliable identification results for mycobacterial clinical isolates.
Authors: Barbara A Body; Melodie A Beard; E Susan Slechta; Kimberly E Hanson; Adam P Barker; N Esther Babady; Tracy McMillen; Yi-Wei Tang; Barbara A Brown-Elliott; Elena Iakhiaeva; Ravikiran Vasireddy; Sruthi Vasireddy; Terry Smith; Richard J Wallace; S Turner; L Curtis; Susan Butler-Wu; Jenna Rychert Journal: J Clin Microbiol Date: 2018-05-25 Impact factor: 5.948