Teeth: An alternative sample for Body Identification
Forensic scientists have been searching for ways and means to expedite the identification of unknown human remains found. This is crucial for family members that have already filed a missing persons case where their love ones could not be located physically. Although the family members may be able to identify through belongings found such as clothes, body tattoo or jewelry, DNA analysis is still required to authenticate that the cadaver is the actual missing family member, especially in cases where the body has decomposed beyond recognition. Teeth have the potential and ability to provide sufficient DNA due to their structure which is composed of an intertwined matrix of hydroxyapatite minerals (HA), collagen and water. The outer layer of teeth, known as enamel, is the hardest tissue in the human body which automatically protects the DNA in teeth from degradation due to harsh conditions such as high temperature and extreme humidity. Hence, teeth are a non-evasive option compared to long bones which will further damage the cadaver. Furthermore, obtaining bones has an adverse emotional effect on some culture and religion. In this presentation, I will concentrate on the different types of teeth varying in age to investigate the DNA yield diversity, as well as the best method used for pretreatment prior to extraction; up to generating a DNA profile on the GlobalFiler PCR Amplification Kit. The results indicated that pulverizing the teeth using the Freezer Mill followed by extracting DNA using the PrepFiler BTA Forensic DNA Extraction Kit on the Applied Biosystems AutoMate Express Forensic DNA Extraction System gave adequate DNA to produce an interpretable DNA profile to be compared with the DNA profile of the Next of Kin.
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(upbeat music)
- Hi, my name is Aydora.
I'm from the Department of Chemistry Malaysia.
Welcome to HIT 2022.
So we are here online again.
Can we to see you guys physically,
hopefully it'll happen next year.
Today my thoughts gonna be on teeth
and alternative sample for body identification.
So a bit about my Department of Chemistry Malaysia,
we are already the agency of mening,
sites and all-day innovations.
So in short, we cater for the science services.
So all the cases that's pertaining to judiciary,
digital law, pertaining to law, will also be sent to us.
On top of that, we do analysis for environment.
We also support trade industry.
And last but not least, we always ensure that research
and quality assurance is upgraded continuously.
Welcome to my lab, the forensic DNA division.
We are the one and only internationally
appreciated lab in Malaysia.
We conduct crime scene investigations,
CSI in the lab, which means that we look
for biological evidence such as blood stains,
seminal stains, trash DNA, bones and everything found
in the investigation of murder, rape,
sexual assault, drug trafficking, kidnapping,
volume crimes and human remains identification.
So these are the labs that does conduct
forensic DNA services in Malaysia.
We have the model labs or the lab,
we also have the boreal site as well.
So in total, there are actually six labs
that is conducting DNA services in Malaysia.
So why do I wanna talk about teeth as an alternative sample
in body identification cases?
This is a collaboration project with audontologists,
Dr. Rabia from the dentist-defacto team,
University of Malaysia.
So before we can actually use this an alternative sample,
we need to cover the all types of teeth from various age.
So with that, I collaborate with Dr. Rabia
to ensure that I am able to get
from different type of individuals.
And of course, this is all consented
and we have ethics approval for this.
At the same time, we work with decompromised
or degraded teeth from casework as well as research.
So basically it's quite difficult,
you know that you have a lot of teeth in a human being,
but somehow or rather it's quite difficult
to get teeth as a casework case.
And this research is solely
for body identification purposes.
So body identification cases,
cases where the body comes in beyond recognition
where the family members aren't able to recognise
their family members basically.
So in short, most of the time it is burn bodies,
mass disasters, bodies is found
after a certain amount of time,
usually due to the unit and whether in Malaysia,
if a body is left in the wild
or left somewhere for more than a week,
most of the time it is beyond recognition already.
So of course we have family waiting
for the body to be released.
So basically the families is around.
So therefore this type of cases is usually need
to be expedited, which means that we need to ensure
that the results can come out within at least one to two days
so that the family members can be pacified,
whether that is their family member or not.
Of course for confirmation of identity.
Currently most of the time pathologies submit
long bones to the femur, which means that the body
will need to be cut up and then both need to be cut out.
So this offer for me I believe that this is an alternative
sample where it is easy to sample out.
You can just pull on the teeth, you know,
and the bodies are left intact,
which means that you don't need to cut out the feet
or the femur then the family members are able to see
their loved ones and of course last but not as faster
to analyse for us because it's smaller
and it's easier to handle.
So these are the body identification cases
that we receive yearly.
We do have a lot even in 2021.
I would say that we are in lockdown for more than six months.
We still have 155 cases and the case is going every year.
I would like to talk about the actual cases
that we have received about body identification.
So this happens in September 2017
where approximately 23 victims were burned
beyond recognition.
This time in 2017, the pathologists didn't take the teeth.
So imagine that we had to work on long bones,
23 long bones, we had to clean it up
and of course we did not have to clean it up.
(muffled speaking)
But if at that time we have able to implement
the usage of teeth as an alternative,
I believe that it can be confirmed earlier
and of course we do not have to work throughout the night
because it's easier to work with the long bones.
And of course there's another case about a trailer crash
at Burnfall Family members.
Of course again we are unable to differentiate
between the mother's son and her two grandchildren.
And again the pathologists cut out long bones
and of course when it's even it's your recognition
but the teeth are actually intact
and if you were to pull out the teeth again,
it is very much easier.
This is a case that I guess is an international case
and everybody's talking about it,
about the one Kalyan mask brace
where they found 106 kelter bodies
basically from the U.S. in Myanmar.
So in this case we did ask for teeth
and at that time of course we tried both
at that time teeth was not an alternative yet as also now
but then again we are able to work with teeth
and compared to bone teeth are able to give better
DNA profile because the animal of the teeth
attempt to coat the teeth off
so that it doesn't get degraded as easily
as the bone samples.
This is another case about a toddler
who was actually found it.
Well she's actually murdered and left in the woods.
We can't find anything.
What is left of her is just a skull and teeth.
And again in this situation we have work with the teeth
and of course the teeth provides us good DNA
and enables us to get a full DNA profile
which then leads to the catching of the perpetrator
and the perpetrator now as waiting to be a sentence.
So teeth, like I said, to the animal is the hardest issue
in the human body hence is able to protect
with a degradation.
Do you have to understand that our mouth is a cavity?
So therefore if this teeth is used to being subjected
to lots of microbiology.
So because of that tooth is on its own design
to ensure that it is able to withstand lots of bacterial
or lots of, you call that harsh environment
that may degrade your DNA.
As compared to the long bones,
that's why if you are talking about the whole body to say
long bones is preferred because long bones tend to have
this hard shell and is able to preserve the DNA.
By comparing long bones to teeth, long bones in our body
is situated in the flesh.
Therefore it is somewhat shielded from any bacterial activity.
So because of that, this is preferred and is equally capable
even more capable in preserving DNA for a long period of time.
At the same time, you know, talking about teeth sampling,
it is less evasive and non-destructive.
You know, this is much easier to pull out a teeth
comparing to cut out your femur.
When you have teeth because it's small,
cleaning it up and the pretty treatment
is very much easier compared to long bones.
So we've worked with 32 teeth from various age
from aging from 18 years old to 63 years old.
So of course, you can't pull out all their teeth, right?
So you're gonna have only one teeth out of a person
and it's gonna be different type of teeth
and some have a good condition, some have carries,
some have feelings.
And I've also attached the weight of the tooth of each tooth
in the inner table where you can see that it varies as well
as up from 0.11 gram to the most,
because the heaviest is two grams.
So because of that, you can see the different weights
of each tooth.
And of course, the concentration of the tooth
after the many extraction is also being put there
for the whole teeth.
In this study, what I did was to extract the whole tooth
because I wanted to see the genetic concentration
of the tooth that is being done.
So because of that, we are able to see
that there's abundance of DNA from tooth.
But then again, that in mind, this is, like I said,
tooth from a person, which we took, right?
But why do I do this is to give some sort,
like a baseline or some sort, like an understanding
that a teeth is able to give very good DNA.
And even that if you were to have a compromised DNA,
you know, the DNA would not be lost as much.
So you have a good abundance of DNA to start with.
So if you have a degraded DNA, it would not be a severe
that you couldn't get any DNA out of it.
And from this study to teeth, there's only one
that is less than two nanogram per microliter.
So for me, that is a good indication
that it is able to be used as an alternative sample
for the identification.
So a bit of what we did with all the tooth.
So each tooth is physically clean.
So we need to wash it, scrub it, scrape it.
So if there's any carries,
or the filling will be removed out and clean, right?
Then the teeth will be grinded using the fissor meal
or each individual teeth, that means for 32,
that means 32 times I need to put it in a fissor meal.
And of course, with low temperature using liquid nitrogen
and this research, because that harden the teeth
and it's able to crush into a pound dewy form.
So if you can see from the picture,
there's a picture of the fissor meal there,
as well as the tooth once is in powder form.
And of course, the DNA is extracted
using the predfiler BTA, which is the chemistry used.
And we are happy to say that the purification
of this sample is using the ultimate express
to ensure that the DNA extract is pristine and clean.
Of course, there is also other methods
where we can opt to use the phenol chloroform
or we can opt to use other methods.
However, in this study,
because teeth is already small on its own,
so we believe that the best extraction method
for teeth will be using a predfiler BTA on the ultimate express.
And of course, we will quantitate the sample
that is being grinded, which is the today's grinded.
And we quantitated using the quant fillet trial
on the QS5, PODC2D5, you know?
And of course, it's good using the PODC2D5
as you are able to get yield.
And from the sample itself,
you can see that none of the sample
have degradation index is all being well and fine.
And that's proof that the sample that we got
is has not gone through any hash or degradation process.
The extracted DNA 45 as well is then put
to the any amplification using the global fillet chemistry
is amplified on the proflex
using the polymer chain reaction technique.
And then we put through the ABI-P500 XL
to ensure that we are able to get good DNA profile.
And of course, the electrophoresis is well and fine.
Using 32 samples, you are able to get it out
within two to three hours.
So basically that is based on the 32 samples.
And like I said, it's all able to get good DNA profile
because it's good condition.
And all of the 32 teeth able to give us abundance of DNA.
Now I would like to talk about the actual case
that we work after having the baseline of the 32 teeth
that we analyze prior.
So this actual case was a man living in a hut on his own
in a remote area in a
the bornial part of Malaysia, which is Saba.
In Saba, this man was living on his own
in a remote area.
He, the hut that he was living was burned down to the ground.
But nobody knew that he was burned down
because like I said, it was he was on his own.
But then again, his son started looking for him
because he hasn't contacted the sun for more than three weeks.
So because of that, the sun went back to look for the father
and found out that the hut has been burned down
and they found pieces or the cadaver burned down.
And this severely burned remains,
even though that the sun knew that that was the father,
but the law in Malaysia stated that
if you unable to recognize a person physically,
you are not allowed to take back the person
and give them a proper blue rail.
Okay, the pathologies or the doctor in Saba decided
to call me and contact me to see whether I'm able
to get DNA profile out of the tooth tooth
or teeth that they found at the,
at the, at the, at the, at the, at the, at the,
at the front front front area.
Why is it that he contacted us here in, in, in HQ rather
than the people in Saba?
Although we have a DNA like in Saba, like I said,
teeth is not really something that is commonly used
for body identification in Malaysia.
Therefore the methods are not tested yet
and not actually used yet.
So we decided to try out to see whether this burn teeth
is able to yield the DNA profile out.
So the teeth is clean and this is the actual teeth
is being grind out to powdery form.
So using piezomile, then we asked my profile a BK.
Like I said, using the method that we have
successfully done up using the teeth early on.
And then of course,
identify using the Gobo file application kit.
And as you can see, there's a complete DNA profile.
It was successfully generated.
You can see there are signs of preferential amplification
where you can see a sloping towards the higher
or the longer DNA chains.
But then again, all in all it is fine
and it's able to give good and interpretable DNA profile.
We are able to match the teeth to the sun.
And we're happy to say that the sun is able
to give a proper burial to his father.
It's always a good feeling to be able to help
or to assist any family members.
And thus giving them closure.
So my recommendation to the study is that teeth
can definitely be used as a sample for body identification.
But make sure you need to ensure that the teeth
is taken from the mouth, the mandible area.
It doesn't count if you were to find tooth away
from the body because then it's gonna be difficult.
Of course, you would say that, hey,
you see this is not something that you can find everywhere.
Yes, but then again, to ensure that that particular person
is the one that you want to identify.
So you need to ensure that the teeth need to be taken
from the cavity or the mandible of that particular person.
It is definitely less invasive for even for the living,
we pull out our teeth.
So therefore, for the non-living,
as well, it is very much better.
And of course, you have no damage to the body.
For your information here in Malaysia,
usually for body identification,
you imagine this is the femur,
they will actually cut the femur.
For the Muslim, especially, they need to bathe the body.
And most of the time is done by the family members.
Of course, it's gonna have some adverse effect
when you see that your loved ones, limbs,
is actually like half not there.
And even more so for the embalming process
where the person doing it also is gonna have
kind of difficult time to ensure
that everything is well and fine.
And furthermore, teeth is able to give good DNA use
and definitely preferred by family members.
For future plans, I'm gonna have some more teeth coming,
lots more teeth coming in from the faculty of the industry.
So, since working with compromise or degraded teeth,
it's quite difficult to get from case one.
So what we plan to do is to actually put this teeth
into very, I call that adverse effect
or to have mock samples where the teeth will be burned
at varying degree.
So we plan to do that.
At the same time, we plan to bury the teeth
in different type of soil at different type of time.
So we hope that this will actually gives us a more
solid research on the usage of teeth
even though it has been put through lots
of degradation process.
And having these teeth protocols in
expedite the current technique
because cleaning a smaller amount of teeth is very small.
It's very much easier than having to work with long bones.
And of course, powdering it takes a bit of time.
So we are trying to see whether
see less loss of DNA on the teeth.
Rather having it into pieces,
you just break up into pieces,
will it be enough to yield the DNA as compact
to pound it that we have successfully done.
So let me end with it.
The law has made sure we next remain a matter of science.
You have no victim to avenge,
no guilty innocent people to reveal or save.
You must back as the money within the limits of science.
With that, thank you so much.
I hope that my research is enabled to help other labs
and other departments in expediting
the body identification.
(silence)
- Hi, so I've got some questions.
The questions that I have is question one.
Have you used other chemistry to extract from teeth?
Yes, we have tried using phenylchorophone.
However, for me, the best so far to extract
will probably be automated from some of the more fisher.
I have not tried using other chemistry
from the bermig
out to the bone and teeth.
But at the moment, I'm happy with what you're using.
Now also I've got questions that,
what were the challenges that I faced
when grinding teeth?
It's not as difficult as a bone.
The teeth is much more easier to grind.
You can just bake it out with a pestle and mortar
and then you put it into the fizzomeal.
And then I've got the third question.
How did you clean the teeth before you extract the DNA?
Cleaning teeth is easy.
You just use the normal,
like for that cleaning the outside layer
and then you, of course, wash it with water.
And then after that, you bake the teeth and put it and grind it.
Have you tried cartilage for the DNA extraction?
Of course, before this, my thought is about how you can actually change
from taking cartilage or even
bones, long bones in order for you not to damage the cadavers
or the human remains.
Using tooth is much easier.
Cartilage, however, you have to look at the age or how long
the human's remains have been left there.
At certain times, if the body has been left there for quite some time,
cartilage is not able to give you good DNA profile.
As compact with teeth,
question five, can you share with tooth is do you think is best?
Based on the 32 teeth that we did,
I would think that it's not as,
it would not be anyone that released any type of tooth that really stands out.
But of course, people would prefer the molar teeth as compact to other teeth.
Can you share this?
Question six, do you be calcified the teeth?
No, no.
We don't be calcified the teeth.
In fact, nowadays, we don't be calcified even the bones unless you're using
phenylchorophone.
A lot of methods that are current methods,
you can actually get DNA out without
the calcifying teeth or long bones or cartilage.
Have you tried getting mRNA from teeth?
No, I have not.
I have not tried.
We are still in the midst of trying to use teeth.
What future plans you have for the teeth research?
Okay, like I said, it's an alternative.
And if it's best, teeth can be used to identify any human remains or body
remains.
If you couldn't get any more blood.
All right.
The police record, the only one minute left.
So I'm going to just answer the questions.
Can I have a copy of your presentations?
Please email me directly.
I'll get to you.
Please, the dental records before passing the teeth to the left.
I'm not sure.
I believe they would get the dental records from the otontologist.
When it comes to me, they have gone through the pathologies and the otont
ologist.
So I believe that the police should have done that as well.
Have you tried teeth sample or rapid DNA?
No, I'm going to try that.
But then again, I probably not think that it worked because for rapid,
you could not have the, we call that defined bones, but we'll try and see if I
can break
it to maybe bigger pieces and put it on the rapid.
What if they bone samples on our human real predators and negative results?
Well, if it's human, it will not be quantitative because my quantitation is
human quantitation
kit.
I've got 10 seconds.
Thank you so much for supporting and thank you so much for all the questions.
Please email me if you need more answers or if you need more publications on
myself.
See you guys again and thank you to all.
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