Comparing the identification of Clostridium spp. by two Matrix-Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) mass spectrometry platforms to 16S rRNA PCR sequencing as a reference standard: a detailed analysis of age of culture and sample preparation

Roy Chean, Despina Kotsanas, Michelle J Francis, Enzo A Palombo, Snehal R Jadhav, Milena M Awad, Dena Lyras, Tony M Korman, Grant A Jenkin

Research output: Contribution to journalArticleResearchpeer-review

Abstract

We compared the identification of Clostridium species using mass spectrometry by two different Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) platforms (Bruker MS and Vitek MS) against 16S rRNA sequencing as the reference standard. We then examined the impact of different sample preparations and (on one of those platforms) age of bacterial colonial growth on the performance of the MALDI-TOF MS systems. We identified 10 different species amongst the 52 isolates by 16S rRNA sequencing, with Clostridium perfringens the most prevalent (n=30). Spectrometric analysis using Vitek MS correctly speciated 47/52 (90.4 ) isolates and was not affected by the sample preparation used. Performance of the Bruker MS was dependent on sample preparation with correct speciation obtained for 36 of 52 (69.2 ) isolates tested using the Direct Transfer [DT] protocol, but all 52 (100 ) isolates were correctly speciated using either an Extended Direct Transfer [EDT] or a Full Formic Extraction [EX] protocol. We then examined the effect of bacterial colonial growth age on the performance of Bruker MS and found substantial agreement in speciation using DT (Kappa=0.62, 95 CI: 0.46-0.75), almost perfect agreement for EDT (Kappa=0.94, 95 CI: 0.86-1.00) and exact agreement for EX (Kappa=1.00) between different days.
Original languageEnglish
Pages (from-to)85 - 89
Number of pages5
JournalAnaerobe
Volume30
DOIs
Publication statusPublished - 2014

Cite this

@article{daafdb42a04441d1854563d8b9dc7180,
title = "Comparing the identification of Clostridium spp. by two Matrix-Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) mass spectrometry platforms to 16S rRNA PCR sequencing as a reference standard: a detailed analysis of age of culture and sample preparation",
abstract = "We compared the identification of Clostridium species using mass spectrometry by two different Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) platforms (Bruker MS and Vitek MS) against 16S rRNA sequencing as the reference standard. We then examined the impact of different sample preparations and (on one of those platforms) age of bacterial colonial growth on the performance of the MALDI-TOF MS systems. We identified 10 different species amongst the 52 isolates by 16S rRNA sequencing, with Clostridium perfringens the most prevalent (n=30). Spectrometric analysis using Vitek MS correctly speciated 47/52 (90.4 ) isolates and was not affected by the sample preparation used. Performance of the Bruker MS was dependent on sample preparation with correct speciation obtained for 36 of 52 (69.2 ) isolates tested using the Direct Transfer [DT] protocol, but all 52 (100 ) isolates were correctly speciated using either an Extended Direct Transfer [EDT] or a Full Formic Extraction [EX] protocol. We then examined the effect of bacterial colonial growth age on the performance of Bruker MS and found substantial agreement in speciation using DT (Kappa=0.62, 95 CI: 0.46-0.75), almost perfect agreement for EDT (Kappa=0.94, 95 CI: 0.86-1.00) and exact agreement for EX (Kappa=1.00) between different days.",
author = "Roy Chean and Despina Kotsanas and Francis, {Michelle J} and Palombo, {Enzo A} and Jadhav, {Snehal R} and Awad, {Milena M} and Dena Lyras and Korman, {Tony M} and Jenkin, {Grant A}",
year = "2014",
doi = "10.1016/j.anaerobe.2014.09.007",
language = "English",
volume = "30",
pages = "85 -- 89",
journal = "Anaerobe",
issn = "1075-9964",
publisher = "Elsevier",

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TY - JOUR

T1 - Comparing the identification of Clostridium spp. by two Matrix-Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) mass spectrometry platforms to 16S rRNA PCR sequencing as a reference standard

T2 - a detailed analysis of age of culture and sample preparation

AU - Chean, Roy

AU - Kotsanas, Despina

AU - Francis, Michelle J

AU - Palombo, Enzo A

AU - Jadhav, Snehal R

AU - Awad, Milena M

AU - Lyras, Dena

AU - Korman, Tony M

AU - Jenkin, Grant A

PY - 2014

Y1 - 2014

N2 - We compared the identification of Clostridium species using mass spectrometry by two different Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) platforms (Bruker MS and Vitek MS) against 16S rRNA sequencing as the reference standard. We then examined the impact of different sample preparations and (on one of those platforms) age of bacterial colonial growth on the performance of the MALDI-TOF MS systems. We identified 10 different species amongst the 52 isolates by 16S rRNA sequencing, with Clostridium perfringens the most prevalent (n=30). Spectrometric analysis using Vitek MS correctly speciated 47/52 (90.4 ) isolates and was not affected by the sample preparation used. Performance of the Bruker MS was dependent on sample preparation with correct speciation obtained for 36 of 52 (69.2 ) isolates tested using the Direct Transfer [DT] protocol, but all 52 (100 ) isolates were correctly speciated using either an Extended Direct Transfer [EDT] or a Full Formic Extraction [EX] protocol. We then examined the effect of bacterial colonial growth age on the performance of Bruker MS and found substantial agreement in speciation using DT (Kappa=0.62, 95 CI: 0.46-0.75), almost perfect agreement for EDT (Kappa=0.94, 95 CI: 0.86-1.00) and exact agreement for EX (Kappa=1.00) between different days.

AB - We compared the identification of Clostridium species using mass spectrometry by two different Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) platforms (Bruker MS and Vitek MS) against 16S rRNA sequencing as the reference standard. We then examined the impact of different sample preparations and (on one of those platforms) age of bacterial colonial growth on the performance of the MALDI-TOF MS systems. We identified 10 different species amongst the 52 isolates by 16S rRNA sequencing, with Clostridium perfringens the most prevalent (n=30). Spectrometric analysis using Vitek MS correctly speciated 47/52 (90.4 ) isolates and was not affected by the sample preparation used. Performance of the Bruker MS was dependent on sample preparation with correct speciation obtained for 36 of 52 (69.2 ) isolates tested using the Direct Transfer [DT] protocol, but all 52 (100 ) isolates were correctly speciated using either an Extended Direct Transfer [EDT] or a Full Formic Extraction [EX] protocol. We then examined the effect of bacterial colonial growth age on the performance of Bruker MS and found substantial agreement in speciation using DT (Kappa=0.62, 95 CI: 0.46-0.75), almost perfect agreement for EDT (Kappa=0.94, 95 CI: 0.86-1.00) and exact agreement for EX (Kappa=1.00) between different days.

UR - http://www.sciencedirect.com/science/article/pii/S1075996414001280

U2 - 10.1016/j.anaerobe.2014.09.007

DO - 10.1016/j.anaerobe.2014.09.007

M3 - Article

VL - 30

SP - 85

EP - 89

JO - Anaerobe

JF - Anaerobe

SN - 1075-9964

ER -