APPLICATIONS OF MALDI-TOF MS IN

CLINICAL DIAGNOSTIC MICROBIOLOGY

Presented by: Dr.Seema Aleem
Moderator: Dr.Varsha.A.Singh

AUTHORS: Antony Croxatto, Guy Prodhom & Gilbert Greub

PLACE: Institute of Microbiology, University Hospital Center and

University of Lausanne, Lausanne, Switzerland

NAME OF THE JOURNAL: Federation of European Microbiological

Societies

VOLUME: 36 (2012) 380407

INTRODUCTION
MALDI-TOF MS: The matrixassisted laser desorption ionization timeof flight mass spectrometry.

HISTORY
1975- Anhalt and Fenselau proposed for the first time that bacterial
characterization could be achieved using Mass Spectrometery .
1980s- Development of desorption/ionization techniques:
-plasma desorption (PD),
-laser desorption (LD)
-fast atom bombardment (FAB)
allowed the generation of molecular biomarker ions from microorganisms
leading to bacterial profiling .

1985- MALDI- Term was coined in by Franz Hillenkamp, Michael Karas.

1996- MALDI-TOF spectral fingerprints could be obtained from whole bacterial

cells without pretreatment before the MS analysis (Holland et al 1996).
2010- MALDI-TOF MS has been used to characterize a wide variety of
microorganisms including bacteria, fungi,and viruses (Giebel et al., 2010).

plasma desorption (PD)

Also called fission
fragment ionization, is a
mass spectrometry
technique in which
ionization of material in
a solid sample by
bombarding it with ionic
or neutral atoms formed
as a result of the nuclear
fission of a suitable
nuclide, typically
the californium isotope 2
52Cf

fast atom bombardment (FAB)

Fast atom
bombardment (FAB) is
an ionization technique
used in mass
spectrometry in which
a beam of high energy
atoms strikes a surface
to create ions.

laser desorption (LD)

Matrix-assisted laser
desorption/ionization (MALDI) is a
soft ionization technique. The sample
is mixed with a matrix material.
Upon receiving a laser pulse, the
matrix absorbs the laser energy and it
is thought that primarily the matrix is
desorbed and ionized (by addition of
a proton) by this event. The analyte
molecules are also desorbed. The
matrix is then thought to transfer
proton to the analyte molecules (e.g.,
protein molecules), thus charging the
analyte.

PRINCIPAL
result (spectrum)
detector

flight tube

laser

Crystallised matrix with

analytes

detection

Separation(TOF)
no electric field

Acceleration
(electrostatic field)
Matrix-assisted laser
desorption ionization

TECHNIQUE
A mass spectrometer is composed of three functional units:
an ion source to ionize and transfer sample molecules
ions into a gas phase,
a mass analyser that separate ions according to their
mass-to-charge ratio (m/z),
a detection device to monitor separated ions.

INSTRUMENT

(MALDI-TOF) MASS SPECTROMETRY:

analysis of blood-borne organisms.

BIOMARKERS
Majority of the biomarkers used in MALDI-TOF are
proteins from the inside of bacterial cells mostly ribosomal
proteins.
These biomarkers have a molecular mass below 15 kDa.
Other biomarkers:
. Nucleic acid-binding proteins
. Cold-shock proteins
. Cytosolic proteins

ADVANTAGES OF MALDI TOF:

MALDI-TOF MS has been used to characterize a wide
variety of microorganisms including bacteria, fungi, and
viruses.
One of the major advantages of using MALDI-TOF
technology for bacterial identification is the time-toresult, which is reduced from 24 to 48 h to less than an
hour.
MALDI-TOF MS is used in applications such as
microbial taxonomy, typing and identification of virulence
factors

PROBLEMS IN MALDI-TOF:

FUNGAL IDENTIFICATION:
MALDI-TOF MS systems for the identification of
microorganisms were successfully adapted for the
identification of fungi in the past 10 years.

Three fungal species, Penicillium spp., Scytalidium spp.,

and Trychophyton rubrum, showed distinct spectral
fingerprints allowing accurate species distinction.

DIRECT IDENTIFICATION FROM

SAMPLES
Given the accuracy of MALDI-TOF for bacterial
identification, this technology might be directly applied to
some clinical samples, such as:
Blood
Urine
Cerebrospinal fluid
Pleural fluid
Peritoneal liquid
Synovial fluid.

LIMITATIONS:
The major limitation is the amount of bacteria present in
the samples and the limit of detection of current MALDITOF protocols.
bacterial identification from cerebrospinal fluid is yet not
applicable in routine diagnostic laboratories,due to low
bacterial load and the limited volume of sample available.