You
You
Do you want to go to the new norm for the control of the meat products in the United States that as I mentioned here the doctor
Enter in vigor from the 4th of June
Well, here is the copy of what is the norm of what I will be looking for that are the top-stacks or the big six that are like
We are knowing here in Mexico
The types of samples that are going to be monitoring that are going to be raw meat
ground meat and base for imported products
Here, well, it is a little bit of the cost of the industry to implement the method both screening as the process for positive lots
The main matrixes by region that well we are going to cover what is this
We belong to the North America part, in the part of the meat, this trimmy, well, that is inside and ground, and well, the part of green leaves
Well, why is it called Ecoli
Top-stack or the big six?
Well, there are more than 50 different types of Ecoli in hemorrhagic
which were identified as causes of diseases in humans, but there are seven that are the main ones
So we call them big six, but we are also including the O-157 that we are currently monitoring
So, well, the O-157, the O-145, the O-26, the O-145, the O-103, the O-111, and the O-121
Why are they pathogenic? Well, what they do is cause hemorrhagic colitis and the O-047 causes hemorrhemic emolytic syndrome
The two infectants are low, well, the studies show that, for example, in the O-126, in Indyna-Marca, only with 100 UFCs, they had problems
The O-111, in Bovina, in Australia, with 10, the O-145, in Belgium, with 400, and the O-157, two infectants, is of 10 UFCs
How are they pathogenic? Well, what they do is cause hemorrhagic colitis, which are STX-1 and STX-2
and they use the EAE mechanism to connect to the wall of the intestine, and that's how the toxin passes through the blood system
The regulatory definition of the topstech is the following, the definition is, it indicates that they are E. coli that present a combination of the O-0 group
and the genes that define the pathogenicity of the E. coli in hemorrhagic colitis
These are the group, the O-0 groups, which are the ones I mentioned, which are the 7
and the genes of pathogenicity are E. A. E., STX-1 and STX-2, and all of this is the definition, they must have the O-0 groups, E. A. E., and STX-1 and STX-2
This is the method of the UFCs, the part of the ingesting
Here it is, well, 325 grams of sample, 995 of average, the average in which it is, the time, well, the temperature, the time
then they have to do a PCR to define the pathogenicity and the second PCR would be to define the O-0 group
and then that it gives them a positive, they have to confirm, and here it is how they would have to do the isolation,
that they have to use the immunomagnetic separation, they have to use the agar rainbow, then they have to do another PCR
and then see if they have E. coli or not, so really this is very late and well, you are industry, you really need results much faster
Now, how to define the analysis method, well, there are two options, we have immunological methods, you must know more than one
but only half of the definition, they are Liza-type methods, lateral flow or alpha-type, the group O is not indicating pathogenicity
many do not have E. coli, they can give reaction, the E. coli with the correct group O can not have one more pathogenicity genes
and also there is a, well, they have a false positive.
And there are also genetic methods, which is by means of PCR, which is only half of the definition,
there are more than 200 E. coli types that have one more of the target genes, many non-E. coli can contain the target genes
there is also a false positive and there is a multi-organism effect that there is no way to determine if all the targets are present in a single organism
Now, the solution that we are offering is to combine the two methods, that is, to have the immunological part and also the genetic part
and that is what we do, we, our method involves the immunomagnetic separation, which is what we are already capturing,
the group O and when we move on to PCR, we are already doing the part of the detection of pathogenicity genes
Here is how it would be, well, in theory what we have to do, we have a kit exclusively to detect the 7, we call it top 7
we do a 18-hour enrichment at 42 degrees in the middle of HEK, then we do the immunomagnetic separation
that here we are already capturing all the O's, the group O's and then we go directly to our PCR team to do the analysis and detect the E. coli genes
or the STX1 or STX2 and also the O157H7, and well, we also have, if we have another kit that detects the 6, in case they do not want to have the O157
in the same result, that the industry in the United States is using the top 7, because they have all the results there
these are all the tests that we have available with our system
and well, the next presentation is the system as such, yes?
Well, as I mentioned, our system is called Genetic Detection System, Shuran's GDS
what we do is we use the latest developments in technology to achieve the best results and precision
we have three areas that are completely different from what is in the market
which is the preparation of the sample that is based on the immunomagnetic separation
we use highly specific probes and primers, and we also have a rotating instrument innovative platform
well, we have advances in multiple areas, we have the preparation of the sample with the immunomagnetic separation
we have a reactive system that comes all the way, ready to be used
and also the instrument platform, therefore we have precision, quickness and easy to use
as for the preparation of the sample, as I mentioned, we use the immunomagnetic separation
well, the specific antibody captures and involves the organisms
the capture is the antibody covered with particles for magnets
and the separations with the magnets collect and go to the particles that cover the organisms
this is called Spick Pen, it is patented
but this is the device that we use to make the immunomagnetic separation
and well, one of the advantages is that we improve the purity of the sample
we reduce the competitive microflora, we have an increase in specificity
we improve the relation of the target organism in boxes or dishes with agar
we improve the selectivity of the enrichments
we are also removing the potential inhibitors of PCR
as they can be food residues, sanitizers or crops
and we are also increasing the amount of DNA for the analysis
because we are concentrating on the sample
so we concentrate on the organisms, we try from 1 to 2 log of increase in density of cells
post-enrichment, therefore we have a greater amount of DNA to analyze
and well, here is how our Spick Pen works
what we do is that the sample and the particles combined are in the sample block
which is part of our method
the capture of particles and the involvement of the organism without resolution
the particles are physically extracted from the sample
the tip is immersed in the dilapidated solution
the resuppended particles and then we go directly to the detection system
once we have our sample enriched
when we use the Spick Pen there is no greater manipulation
we capture it, we make a dilapidated solution
and then we transfer it to the amplification tubes
as for the reactive system, I mentioned that everything is well dilapidated
ready to be used
this is here a little more or less how the whole reaction of PCR is carried out
here is the part of the amplification
the components that we require, which would be the target
which would be what we are trying to detect
which is a specific sequence of the DNA chain
of the organism that we are trying to detect
well, what is the DNA?
the sequence that is amplified by the PCR process
the primers, which is the simple chain of the sequence of the DNA found in the target
they are responsible for initiating the copy of the sequence
they are used in pairs, obviously we need the polymerase attack
which is a thermically stable enzyme responsible for copying the DNA target
and the nucleotides, which are the DNA contractor blocks
this would already be one of the cycles of PCR
we have the first step, which is the denutrification of the DNA
which is a prosthotermic
first we separate the two chains when we heat up to 95 degrees
then the primers are one of the complementary sequences
because we are cooling at 55 degrees
then it begins to work the polymerase attack
and we are developing new chains and we are repeating the cycle
as for the detection, we use a probe
for the detection of the microorganism
or the good of the DNA
then we have a co-existent detection amplification
specific sequence, additional specificity
it is not affected by external primers
nor by a strange DNA
and there is a discreet signal for the target and the internal control
then the benefits of our reactive system
which is still available
is that with the probe we are improving the specificity
since it allows shorter sequences
prevent unwanted unions
and we also have a better discrimination of the target
we do not interpret fusion curves
therefore we are saving more than an hour in time of instrumentation
and there is no subjective interpretation
the moment the probe detects the DNA
it is doing a graph and every time it crosses the control line
we consider a positive
here is how the results are presented
we have a specific detection of the target
inside the reactive system we also have an internal control
inside each of the tubes we have an internal control
then this one from the left side
is an example of graphic results of samples
I mentioned that any line that crosses is considered a positive
and on the right side we have a graphic of the internal control
what we do is to know that the reaction is carried out
regardless of our negative results
normally the industry has negative results
we are always waiting for negative results
so to guarantee that the reaction is carried out
we have the internal control
and here it always has to be amplified
each line is a sample number
and it is of a different color
depending on the number of samples you put in the cycle
that number of lines you have
and they all have to be amplified
we have internal control
a real game of independent probes, separate signals
and there is a direct detection of the target
reaction 1 to 1 is the target
and it would be the platform of the instrument as such
I mentioned that it is a rotating system
here we are comparing with traditional thermos
that would be a traditional format
what they do is they use a block of pellet
based on a cooling heater
and it has the detection on a single channel
here it has the limitation that they cannot guarantee
that all the samples have the same temperature
and normally they do not use the external samples
because it is a thermal reaction
and they cannot guarantee that it is carried out
inside each of the tubes
so they have several limitations
the slow cycles that increase the temperature
cycles with inconsistent temperature between the tubes
and they are also dependent on a colorant
not specific that is the cybergreen
and also on the analysis of fusion curves
what we are
the samples are accommodated in a rotating format
as I mentioned for direct heating and cooling
we have centrifugal movement with constant air exchange
the heat transfer with our system is by convection
and in the block of pellets by conduction
it ensures uniform temperature through all the reactions
and we eliminate long periods of temperature
required by the systems based on blocks
so here we are making a comparison
against the rotating system that is our system
comparing with the block
this is only the time of instrument
no preparation to show
and only instrument
so here we see time when the neutralization begins
3 minutes, 10 minutes
time left in a cycle, less than 2 minutes, 4 minutes
total time of 70 to 170 minutes
fusion curves we do not interpret here is
minimum 60, so comparing only
or rotating block is saving 2 hours and a half
of time of results
that finally for the industry
the faster you get the result
the faster you can release your lotus
or take corrective actions
and here would be the advantages of the team
as I said they are three areas
preparation of the sample, reactive themes
and instrument platform
speed, with the non-magnetic separation
or the peak pen, we concentrate the target
therefore we have shorter requirement time
specificity, with the non-magnetic separation
we have specificity of the first level
easy to use because we prepare the sample
in just 20 minutes
all the samples in just 20 minutes
and with precision we separate
the target of the levels of the PCR
as for the reactive system
we are with the use of the zone
we eliminate the fusion curves
in terms of specificity
only amplify the target
and the probe only detects it
easy to use because the reactive
is used in the amplification tubes
and in precision we have
specificity of primers and probes
and as for the instrument
thermal efficiency, we reduce cycle time
multi-channel system allows the use
of high specific probes
final results, since we don't have
to interpret fusion curves
and we have a consistent PCR
since the reaction is carried out
by amplification tubes
and this is what we saw
because I started with that
this is the same part
of the STEC
if you have any questions
comments
we need three areas
but not separate
to compare
we need the area 1
where we add all the reactive
which is a clean area
area 2
where we add the sample
and the area 3
which is where the team is
but the team is small
you don't need much space
to implement it
the same as you
implement good laboratory practices
there is no risk
to use gloves, you have to use a bat
you have to be careful
but that enters into the good laboratory practices
that part
we see it right now
because we don't sell directly
through our distributor
which is present here too
we have an objection card
by the FCIS
and we are waiting
for your answer
this depends
on the agreement
of the review that is done
with the FCIS authorities
you will see if this method
is used
in Senapa
as a quick method
to obtain the results
and only while
the method
is implemented
by FCIS
is validated in our country
we will be advancing
in validating the method
to the part that
if this is determined
it could be used
this method to start
the sample
of the Shiga toxin
this norm
on ECOLE and STEC
for all types of processes
ready for consumption
cooked
no
it is for raw
it is ground beef
or ground beef
there are reports
of these bacteria in Mexico
not that it is registered
the results obtained
if the methods are approved
at international level
there are
current samples
and external laboratories
there are
primers from Salmonella
ECOLE generica
of mesophiles
Listeria
for this team
we have particular patogenes
of indicators
we have other types of methods
and the Carnic industry
in the United States is using
our system
and last year
maybe 6 to 7 months
since the boom started
with the top 7
and they are using the main
Carnic industry
when detecting DNA
the result includes
microorganisms, living and dead
if so
it is representative
what is the cost
of purchase
in terms of our team
we do not detect dead cells
because we have
a magnetic separation
the only thing we capture are living cells
other PCR teams
detect dead cells
but not only living cells
the new serotypes
or what species do they apply
what type of meat
or
specifically meat
if the product is not
ready for consumption
but if it is an ingredient
for this type of product
it is mandatory
although it carries a posterior process
it does not apply
it would be for raw meat
raw meat
the rule for that tech
was implemented
it is implemented on June 4
in the United States
by the FSI
what is the date of the date
for the FSI to be implemented
the technology
that the FSI has already implemented
in its official laboratories
well from June 4
it will be starting to show
the boats
that arrive from Mexican meat
is
if
it takes a while
to implement the technique
but above all
to validate the method
that is why these additional techniques
are being seen
to detect
both us as an official part
as you
as the companies
the laboratories have to see
the convenience of the teams
and
if there was a positive case
after June 4
it would have
the next boats
of this company
not counting the result
for the specific SEPA
and that tech in the product
is a limiting to export it
and for what species it applies
raw
ground
and
if it is going to be a limiting
and
in a given moment
it is going to stop the boats
of the company
if it exports trimmings
it would have to show
all
or how many slots of the load
do you have something already
Aurelio regarding
ah ok, you are going to talk
about it
applies for pork
ground beef
very good
very good
thank you very much
thank you very much
