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MALDI-TOF MSInnovation with Integrity

MALDI Biotyper®

Mycobacteria Identification

The genus Mycobacterium includes, as major groups,the important pathogens of the Mycobacteriumtuberculosis complex (MTC) and the nontuberculousmycobacteria (NTM). Members of the latter groupare cited increasingly as the cause for opportunistic infections among immuno-compromised patients. This trend and the rise of antibiotic resistance in this genus demand improved differentiation among Mycobacterium species.

The MALDI Biotyper mycobacteria suite is the com-prehensive solution for laboratories in need of highly reliable and fast mycobacteria identification via MALDI-TOF mass spectrometry. The Mycobacteria Library 4.0 covers 159 of the currently known 169 Mycobacteria species. 880 strains - of which more than 450 are clinical isolates - cover the natural variability of Mycobacterium species. The MBT Mycobacteria SW Module with adapted data acquisition and analysis secures high sensitivity of mycobacteria identifications.

Figure 1: Mycobacteria samples prepared with the MycoEx preparation protocol reveal excellent mass spectra securing sensitive and reliable mycobacteria identification.

MycoEX – the optimized and safe extraction protocol

Mycobacteria are rigid microorganisms for which direct transfer and conventional extraction methods might not yield MALDI-TOF mass spectra of sufficent quality for species identification.

Bruker has therefore developed an easy and safe silica bead-based extraction protocol, the results of which are high-quality mass spectra for species identification (see figure 1).

Mass Spectra of Mycobacterium gastri DSM 44505T

MycoEX method

Images on front page and page 2: Coloured scanning electron micrograph (SEM) of Mycobacterium smegmatis bacteria.

One workflow for solid media and liquid cultures Spectra of mycobacteria grown on solid Löwenstein-Jensen or BACTEC™ MGIT™ (BD Diagnostics) liquid media show no significant variation.

International mycobacteria consortium The aim of this consortium is to collect securely identified mycobacteria strains to create a library as the basis for the highest performance in mycobacteria identification.

Mass Spectra of Mycobacterium peregrinum DSM 43271T

Figure 2: Mass Spectra of Mycobacterium peregrinum DSM 43271T cultivated on Löwen-stein-Jensen medium (top) and in MGIT™ tube (bottom), using the MycoEX protocol.

Twenty-five laboratories spread across10 countries have provided well characterized clinical strains from which more than 450 reference entries have been derived, improving significantlythe strain coverage for clinical analyses.

Mycobacteria Library

159 species entries

M. abscessus subsp. abscessus M. cosmeticum M. komossense M. phlei

M. abscessus subsp. bolletii M. crocinum M. koreense M. phocaicum

M. africanum M. diernhoferi M. kubicae M. porcinum

M. agri M. doricum M. kumamotonense M. poriferae

M. aichiense M. duvalii M. kyorinense M. pseudoshottsii

M. algericum M. elephantis M. lacus M. psychrotolerans

M. alvei M. engbaekii M. lentiflavum M. pulveris

M. arabiense M. europaeum M. litorale M. pyrenivorans

M. aromaticivorans M. fallax M. llatzerense M. rhodesiae

M. arosiense M. farcinogenes M. longobardum M. riyadhense

M. arupense M. flavescens M. madagascariense M. rufum

M. asiaticum M. florentinum M. mageritense M. rutilum

M. aubagnense M. fluoranthenivorans M. malmoense M. salmoniphilum

M. aurum M. fortuitum subsp. acetamidolyticum M. mantenii M. scrofulaceum

M. austroafricanum M. fortuitum subsp. fortuitum M. marinum M. sediminis

M. avium subsp. avium M. fragae M. marseillense M. senegalense

M. avium subsp. paratuberculosis M. franklinii M. microti M. senuense

M. avium subsp. silvaticum M. frederiksbergense M. minnesotense M. seoulense

M. bacteremicum M. gadium M. monacense M. septicum

M. boenickei M. gastri M. montefiorense M. setense

M. bohemicum M. genavense M. moriokaense M. sherrisii

M. botniense M. gilvum M. mucogenicum M. shimoidei

M. bourgelatii M. goodii M. murale M. shinjukuense

M. bovis M. hippocampi M. nebraskense M. simiae

M. branderi M. gordonae M. neoaurum M. smegmatis

M. brisbanense M. haemophilum M. neworleansense M. sphagni

M. brumae M. hassiacum M. nonchromogenicum M. stomatepiae

M. canariasense M. heckeshornense M. noviomagense M. szulgai

M. caprae M. heidelbergense M. novocastrense M. terrae

M. celatum M. heraklionense M. obuense M. thermoresistibile

M. celeriflavum M. hiberniae M. pallens M. tokaiense

M. chelonae M. hodleri M. palustre M. triplex

M. chimaera M. holsaticum M. paraense M. triviale

M. chitae M. houstonense M. paraffinicum M. tuberculosis

M. chlorophenolicum M. immunogenum M. parafortuitum M. tusciae

M. chubuense M. insubricum M. paragordonae M. vaccae

M. colombiense M. interjectum M. parakoreense M. vanbaalenii

M. conceptionense M. intermedium M. parascrofulaceum M. vulnerans

M. confluentis M. intracellulare M. paraseoulense M. wolinskyi

M. conspicuum M. iranicum M. parmense M. xenopi

M. cookii M. kansasii M. peregrinum

The Best Mycobacteria Identification

The comprehensive library in combina-tion with optimized software – including adapted thresholds for Mycobacterium ssp. – ensures highly sensitive and specific identification in an fast and easy workflow.

These benefits are offered in a cost effective suite to ensure the highest quality mycobacteria identification solution ever.

Figure 4: Results can be exported easily in a clearly structured and comprehensive report.

The resultant report for each sample shows the two top matches along with their respective matching score. The top ten matches, along with precise information about the closest matching strain, can also be displayed (as shown in figure 5).

Identification results

Figure 3: Easy selection of the sample type during sample preparation for optimized data acquisition and analysis.

Sample Type

Figure 5

Sample 2

Sample Name: A3Sample Description:Sample ID: A3Sample Creation Date/Time: 2016-02-24T13:06:44.502Sample Type: Mycobacteria sampleIdentification Method: MALDI Biotyper MSP Identification Standard Method 1.1Preprocessing Method: MALDI Biotyper Preprocessing Standard Method 1.1ACQ Method: D:\Methods\flexControlMethods\MBT_FC.parACQ Timestamp: 2016-02-24T13:11:07.758Applied MSP Library(ies): Mycobacteria Library (bead method) / 479163c1-0ea8-46a6-8ccc-70422d833fac /

2016-02-23T11:09:06.169

Rank(Quality) Matched Pattern Score

Value NCBI Identifier

1(+++) Mycobacterium phlei 3614_09 FZB b 2.20 123624423

2(+++) Mycobacterium phlei DSM 43239T DSM b 2.08 123624423

3(+++) Mycobacterium phlei DSM 43073 DSM b 1.96 123624423

4(+++) Mycobacterium phlei DSM 43470 DSM b 1.93 123624423

5(+++) Mycobacterium phlei DSM 750 DSM b 1.92 123624423

6(+++) Mycobacterium phlei DSM 43214 DSM b 1.86 123624423

7(+++) Mycobacterium phlei 3614_09 FZB b L 1.81 123624423

8(+++) Mycobacterium phlei DSM 43072 DSM b 1.81 123624423

9(+) Mycobacterium phlei DSM 43071 DSM b 1.79 123624423

10(+) Mycobacterium phlei 9652_00 FZB b 1.72 123624423

Run Identifier: 160224-1303-101003846 Run Creation Date/Time: 2016-02-24T13:06:44.465

Report created at 2016-02-24T13:41:20 Research Use Only Page 10 of 61

Adopted thresholds for increased sensitivity

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MALDI Biotyper® is a registered trademark of Bruker Daltonik GmbH in the European Union and USA.

For research use only. Not for use in diagnostic procedures.

Bruker Daltonik GmbH

Bremen · GermanyPhone +49 (0)421-2205-0 Fax +49 (0)421-2205-103

Bruker Daltonics Inc.

Billerica, MA · USA Phone +1 (978) 663-3660 Fax +1 (978) 667-5993

ms.sales.bdal@bruker.com - www.bruker.com

Order information

Part-No. 1829016MBT Mycobacteria library –including license for the MBT Mycobacteria SW Module.Pre-requisite for the module is the MBT Compass SW.