ERP Micro December 2019
( ee ) Trifluoroacetic acid. ( ff ) Colony-transfer device. ( gg ) Inoculation loops.— Sterile. ( hh ) Microcentrifuge tubes .—1.5 mL.
Biotarget 96 (Cat. No. 1840375) with the MSP adapter MALDI Biotarget (Cat. No. 8267615). ( j ) HCCA (α-cyano-4 hydroxyinnamic acid) matrix .—Portioned (Cat. No. 8255344) or MBT Galaxy HCCA Matrix GPR (Cat. No. 1823405). ( k ) Internal positive E. coli DNA .—Bacterial Test Standard (BTS; Cat. No. 8255343). Additional items required: ( l ) Incubators.— Capable of maintaining temperatures of 34– 38°C for optimal growth. ( m ) Pipets .—Adjustable, variable volume, capable of sampling and delivering 1–1000 µL. ( n ) Micropipet tips.— Aerosol-resistant. ( o ) Refrigerator.— Capable of maintaining 2–8°C. ( p ) Freezer.— Capable of maintaining –20°C. ( q ) Tryptic soy agar with yeast extract (TSA/YE) .—Standard formulation. ( r ) Oxford agar (OXA). —Standard formulation. ( s ) Modified Oxford agar (MOX). —Standard formulation. ( t ) Ottaviani and Agosti agar (O&A). —Standard formulation. ( u ) RAPID’ L.mono (RLM) agar.— Bio-Rad. ( v ) PALCAM. —Standard formulation. ( w ) Vortex mixer. ( x ) Bench-top microcentrifuge.— Capable of 15 871–21130 g , equivalent to 13000 to 15000 rpm for Eppendorf tube centrifuged with a 5424R rotor. ( y ) Standard solvent.— Acetonitrile 50%, water 47.5%, and trifluoroacetic acid 2.5%. ( z ) Water .—HPLC grade. ( aa ) Formic acid. ( bb ) MacFarland standards. ( cc ) Absolute ethanol. ( dd ) Guanidine hydrochloride (GdnHCl).— 8M, commercially available.
( ii ) Fiber-free cloth .—For cleaning MSP 96 target polished steel BC (Cat. No. 8280800) or MSP 48 target polished steel BC (Cat. No. 8281817) only. C. General Instructions Follow all instructions carefully. Failure to do so may lead to inaccurate results. Caution.—Laser class .—The MALDI Biotyper System with a closed safety cover is a Class 1 Laser (exempt from all beam hazard control measures) product. With the safety cover opened, it becomes a Class 4 Laser (considered to be hazardous to the skin and eyes from direct and scattered radiation) product. It has been designed and tested in accordance with International Electrotechnical Commission (IEC) Publication 1010-1 (Pollution Degree 2) Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use . Do not use the MALDI Biotyper System until it has been installed by a Bruker service representative, and laboratory staff have been appropriately trained by a Bruker representative. Read the user manual for the Microflex LT/SH Smart (No. 605415) or the Microflex (No. 1831173) for operational and safety instructions before using the MALDI Biotyper System. Refer to all user manuals below for complete safety precautions: ( 1 ) Instructions for Use MBT Galaxy HCCA Matrix GPR when using the Galaxy to support the sample preparation ( 2 ) Instructions for Use MBT Biotarget 96 ( 3 ) Instructions for Use Bruker Matrix HCCA ( 4 ) Instructions for Use Bruker Bacterial Test Standard ( 5 ) User Manual MBT Subtyping Module ( 6 ) User Manual MBT Compass
Table 2017.10B. Summary of results for the confirmation and identification of Listeria species (non- monocytogenes ) : Inclusivity
MALDI Biotyper correctly identified to
Reference correctly identified to
Not tested
Total
Species level
Genus level
Species level
Genus level
MALDI Biotyper incorrectly identified
MALDI Biotyper
MALDI Biotyper
Organism
Reference
Reference
L. ivanovii Ad1737 L. welshimeri Ad1175 L. grayi Ad1504 L. innocua Ad1675 L. ivanovii Ad1308 L. ivanovii Ad1768 L. innocua Ad1676 L. welshimeri Ad1204 L. welshimeri Ad1211 L. seeligeri Ad2634
60 61 62 60 60 59 60 61 62 59 62 60
61 61 62 62 62 62 62 62 62 62 62 62
15 15 16 14 16 12 12 16 16
16 16 16 16 16 16 16 16 16 16 16 16
0 0 0 0 0 0 0 0 0 0 0 0 0
0 0 0 0 0 0 0 0 0 0 0 0 0
3 3 2 2 2 2 3 2 2 2 2 2
61 61 62 62 62 62 62 62 62 62 62 62
16 16 16 16 16 16 16 16 16 16 16 16
9
L. grayi Ad2064
15 10
L. seeligeri Ad2637
Total isolates
726
742 a
166 b
192 c
26
742
192
a MALDI Biotyper result identified to a genus level. b Total number identified to the species level by the reference method from TSA/YE. c Total number identified to the genus level by the reference method from TSA/YE.
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