BOOM

bioMérieux’s proprietary technology

Extraction Principle

Marina Perrotte, 08/2007

 Nucleic acid extraction
procedure
Various methods have been described for the release and/or extraction of
nucleic acids from a wide variety of clinical specimens.
The purpose of these methods is to:

disrupt infected cells and microorganisms, resulting in the release of nucleic acids

protect the nucleic acids from degradation

remove inhibitors of amplification

concentrate the target nucleic acid

recover nucleic acid in an environment suitable for use in polymerase chain reaction (PCR)

The complexity of the method chosen will depend on several factors. These include:

type and volume of specimen to be tested

concentration of target nucleic acid present/volume of specimen

nucleic acid sought (DNA and/or RNA; single stranded vs double stranded)

presence of inhibitors of the PCR which may interact with the target nucleic acid and/or
the polymerase enzyme

facilities available

safety requirements

Frank SIMONS | Slide 2

Marina Perrotte, 08/2007
 Supports

Different commercial supports exist for Nucleic Acids extraction

Commercial kits can be specialized for DNA and/or RNA extraction

 Liquid reagents

For high specimen input volume

Centrifugation

Automated procedure
 Columns with Membrane or Silica

Format individual or 96-well microtiter plate

Aspiration or centrifugation

In 96-well microtiter plate, can be automated
 Silica particles (non Magnetic)

Centrifugation

Manual
 Magnetic Silica particles

Magnet use

Automated procedure
Frank SIMONS | Slide 3
Marina Perrotte, 08/2007
 BOOM
Extraction Method

Samples lysed in lysis buffer containing chaotropic agent: guanidine thiocyanate (GuSCN).
Cells, bacteria, and viruses present in the samples are lysed and proteins such as nucleases
are denatured and inactivated

Nucleic acids (RNA and DNA) are released and bind to silica particles

Following several washing cycles using wash buffers.

In the final step, the nucleic acids are eluted from the silica particles in an elution buffer

Eluate is highly purified and concentrated nucleic acid for subsequent amplification,
sequencing and other molecular diagnostic analyses.
Frank SIMONS | Slide 4
Marina Perrotte, 08/2007
 BOOM
Licences
Different companies have a licence for BOOM
technology use and to be allowed to sell their
Nucleic Acids extraction methods

N
IO
D T
A R AC
M ND XTR
O TA E
O
B S DS Most of them are competitors for bioMérieux:
LD CI Roche
O
G IC A Bayer
H E E Qiagen
T C L
U Promega
N Toyobo
R
FO Takara
Eppendorf
Diagnocure
Visible Genetics
Frank SIMONS | Slide 5
Marina Perrotte, 08/2007
 BOOM Next generation
Magnetic silica vs.
non-magnetic silica

1 µm
1 µm

magnet
magnetic silica non magnetic silica

silica • better surface / weight ratio: higher binding capacity

• easier to manipulate and automate
Frank SIMONS | Slide 6
Marina Perrotte, 08/2007
 Magnetic silica
Efficient mixing

magnet

Marina Perrotte, 08/2007

silica Frank SIMONS | Slide 7
 Magnetic silica
Efficient mixing

magnet

easyMAG
disposable

Frank SIMONS | Slide 8

Marina Perrotte, 08/2007
 Magnetic Silica particles
Advantages

Frank SIMONS | Slide 9

Marina Perrotte, 08/2007
 Magnetic Silica particles
Advantages
High Purity & Recovery

Eluate highly concentrated in DNA and RNA for a wide range of
downstream applications, to detect low concentrated targets

Eluate with high quality, without any inhibitors for efficient and
accurate, sensitive detection of targets

Rapid, Easy & Flexible

Only one set of reagents

Easy Automation with Magnetic silica particles

Rapid and Efficient Washing steps

Magnetic process allows:
♦ High Throughput Routine Process
♦ Workflow optimization
Optimization
Frank SIMONS | Slide 10
Marina Perrotte, 08/2007
 Frank SIMONS | Slide 11
Marina Perrotte, 08/2007