DNA Testing / Screening for Amyloidosis
I am taking time to document this because several breeders have asked the same question. This was Armageddon 3 for
me. #1 was when my entire OLH breeding program was wiped out by amyloidosis. #2 was shortly after I had started
screening kittens for cardiomyopothy and just for a lark I decided to screen ALL 4 or 5 litters, not just the one litter out of
known carriers and discovered that a major number of my Bali/Javi adults had severe cardiomyopothy. #3 began this last
4th of July with the death of a girl that pathology (red congo stain) showed was due to amyloidosis of both kidney and
liver. Not helping my stress level was the knowledge that a dear friend had just recently lost several cats (distantly
related to the father) to amyloidosis. I also discovered that the mother of my girl had died following her 3rd litter and
that 2 of the mother’s siblings had also died of liver failure. I pretty much came unglued as I have NUMEROUS cats
related to both parents in one way or another. I prepared for the worse and proceeded in much the same manner as I
had during Armageddon #1, with a few exceptions.
CFA is now heavily promoting the DNA project http://CatGENES.org For $39- you can get a DNA profile of your cat and as
new tests are added, the price will not increase. They also “bank” any material not used during testing for future
research and (I assume) to re-run as new tests are added. I attended the seminar at the World Cat Congress and the
representatives made a point of trying to talk to as many breed council secretaries as were present during the two day
show. During the seminar they said that among the tests that they were working on were dilute and amyloidosis. When
they asked me what I felt was important to my breeds, I replied amyloidosis and cardiomyopothy. “Nice to have” would
be a test for LH/SH… that this test would benefit other breeds as well, such as the Exotics. Not too long after the
Congress, there was an announcement that the test for the Dilute gene was ready AND that they had identified 3 out of
the 4 markers required for LH. The #3 marker is breed specific to the Ragdolls and Maine Coons.
PREVIOUSLY I had participated with various studies that Leslie Lyons had conducted, so I was already trained as to what
sort of paperwork they might require when I contacted CatGENES.org re: samples.
1) I went through my database and identified EVERY cat I wanted to test and/or had tissue samples from prior
posts that would be submitted. As far as testing went, my criteria was as follows: A) EVERY cat remotely related
to the cat that had just died and/or that had any ancestors tagged as having died of liver failure or amyloidosis.
B) Any stud not included in group (A) was tested as a simple precaution.
2) I created a special pedigree form and made sure that I had ALL heath notes entered for 4 generations. This
included notes re: date of death, notes from posts, info re: deaths of siblings – anything/everything possible. A
few examples:
A) Clean EKG – seems to carry 2 variations of cardiomyopothy; from lines w/ amyloidosis
B) Jan-2002 - amyloid in thyroid; seizures (litter sister died of amyloid in liver)
C) Nov-2004; liver (amyloidosis carrier)
3) I created address labels with registered name, CFA Reg#, DOB, Description, Sire x Dam – these labels went on
the DNA samples – Zip lock bags were required for the multiple blood tubes sent to CatGENES.org
4) I created an EXCEL worksheet with the same info as the address labels, also date of sample or date of death.
General notes such as “blood/pedigree for DNA profile submitted” and a column for Genetic notes such as
“from lines with amyloidosis”. The Excel worksheet was enclosed with the paperwork and samples as well as
eMailed to the lab as is was too wide to print in entirety on legal size paper.
5) Final step… vet appointments. Copies of all lab reports (when received) were stapled to the appropriate
pedigree prior to being sent to the CatGenes.org project. I called and checked with my vet… While she ran
panels #25 or #46 on each of my cats, she indicated that each lab has different codes for these blood panels. The
key thing to tell your vet is that you are checking KIDNEY AND LIVER FUNCTION. These are the 2 organs most
commonly hit by amyloidosis… kidney more often in the Abys, liver and spleen more often in the Slinkies – but
Page | 1 Kris.Willison_KLM@usa.net 281-804-7801 6-Aug-2008
my girl that just died had it in both liver and kidneys and as you can see from above, I have pathology (more
extensive than usual) where it was found in the thyroid. It can occur in ANY organ, but if it is a critical organ it
will kill. OTHER signs… often the teeth are in horrible condition – stomitosis is often seen (in Slinkies) in
combination with amyloidosis. Liver hemorrhage (at time of death) is also suspicious and samples should be
taken.
6) For simple DNA testing, when you order a test kit online, they send you bucal swabs and cardboard tweezers for
a hair sample (plus instructions). I figured that if I was going to do this, I might as well do it right… draw enough
blood for research as I planned on DNA testing majority of my cats anyway, just not this many at once.
------ Original Message -----Received: Wed, 16 Jul 2008 04:40:45 PM CDT
From: Info <info@dnadiagnostics.com>
To: kris.willison_klm@usa.net (Kris Willison)
Subject: Re: CatGENES Comments
We can use blood and will need a purple top or EDTA tube. Red top is clotted and makes extraction more
difficult. We do not need much blood at all unless we are to keep the samples for research purposes. If it is just
for testing, then we need 1 ml or less. If these cats are to be used for gene research, then we would like to
have a 7 or 10 ml tube. If any of the cats die of the disease, we also could use some affected tissue.
Best wishes,
Fay Webb, Analyst
DNA Diagnostics dba Shelterwood Laboratories
P. O. Box 455, 569 Bear Dr.
Timpson, TX 75975
Phone: 936-254-2228
Fax: 936-254-9286
e-mail: info@dnadiagnostics.com
http://www.dnadiagnostics.com
Received: Wed, 20 Aug 2008 10:21:49 AM CDT
From: DotsRNB@aol.com
To: kris.willison_klm@usa.net
Subject: Re: Brain Storm - Bali/Javi: amyliodosis
> Kris
>
> I talked to Dr. Cothran at the Texas A&M genetics research lab yesterday.
> He is one of the top DNA researchers in the world. We talked about samples
> and the best source of DNA. He suggested that the best samples would be
tissue
> either frozen and sent with a cool pack, or physiological saline sent the
> same way. Tissue that has been fixed with formaldehyde will lose its DNA,
and
> may not have an adequate source for research purposes. Blood is another
Page | 2 Kris.Willison_KLM@usa.net 281-804-7801 6-Aug-2008
> source, and a good one, but not as good as tissue. The blood should be in
EDTA.
I am more than willing to assist anyone else in the paperwork for this endeavor. I can create PDF files w/ pedigrees and
address labels that you can print yourself.
There are tax free funds established for research, but without samples the research can never be completed. They are
interested in samples from cats for both amyloidosis and cardiomyopothy. I am sure that they would not turn down
lymphosarcoma either. I don’t think that blood panels are going to be any help screening for cardiomyopothy. My vet
has a portable EKG and some cat shows have HCM clinics, so if you are aware of carriers and/or loose a cat to any of
these diseases… PLEASE forward a complete pedigree and tissue samples to the CatGENES.org project.
PS – lesson learned, take TWO tissue samples if having pathology run locally. Took two days for my vet’s techs to find
the right body and thaw her enough to get the second sample to send off to the DNA lab.
Good news, where the 1st Armageddon resulted in total devastation… only 1 cat that was heterozygous rather than
homozygous and is still alive and well in a pet home. This time around the only fatalities were the 1st cat and my
retirement fund.
Added notes, 8-August-2008:
The ONLY sure way to confirm amyloidosis is with a lab test known as CONGO RED STAIN to locate the amyloid deposits
with in the organ. While it is possible to do a needle biopsy on a live animal, it is not practical... there is the danger of
hitting a blood vessel and it would be about the same as hunting for a worm in an apple with a large bore needle - one
shot and blindfolded. If you only get flesh/apple - does that mean that there are no worms or seeds? No, it only means
that you missed your target.
The purpose of running the blood panels is to screen for cats that are obviously in liver or kidney failure. You would not
want to be using them in your breeding program anyway, and these would be the most likely to actually have
amyloidosis. Only pathology run on the organs (after death) is going to confirm one way or the other, and I have one
documented case of amyloid deposits found ONLY in the thyroid gland - cat died from seizures, litter sister died of
hepatic amyloidosis. No... the thyroid gland is not usually an organ I have tested. This was a cat co-owned with my vet
that we had already spent $1k on lab work prior to death with no answers and had full nose to toes pathology run to try
to figure out what went wrong - never did get a good answer.
A clean blood panel only means that the cat is healthy, something very good to know it you are going to use it in your
breeding program. It does not rule out the possibility that it could be a carrier. This is where we need to study pedigrees
and thoroughly document deaths. I initially went through my database and marked all DOCUMENTED amyloidosis
deaths and proceeded to mark their parents and 1st generation offspring as carriers, their full siblings as possible
carriers. Where there is a pyramid of deaths to liver failure... ALL descendants were marked as possible carriers. There is
a 1 in 4 chance that 2 carriers will produce a "clean" kitten and a 1 in 4 chance that they will produce one that will
eventually die of amyloidosis. I am now going back and marking SUSPECTED amyloidosis deaths, liver hemorrhage is
classic, I found the old blood work from Angel from Heaven... almost illegible because of the type of paper, but her liver
values WERE off - enough for me to now mark her as unconfirmed amyloidosis as I try to trace a path from past to
present. I am treating ANY deaths to liver failure as suspicious, trying to map out which cats/pedigrees might be safer to
breed to one another.
Page | 3 Kris.Willison_KLM@usa.net 281-804-7801 6-Aug-2008
Using notes from other breeders, I am marking additional amyloidosis deaths and carriers. The end result is to be able to
generate a pedigree on a given cat and guess the probability of it being a possible carrier and weigh the risk of breeding
to possible mates. The more "?" in the pedigree, the more unknowns... one can only hope that the gene has been bred
out.
The Bali/Javi gene pool is too tiny to just throw up our hands and give up, but if we are honest with one another and
document as much as possible, we might be able to hold out until a DNA test is available. Inbreeding, doubling back on
certain lines will be our downfall, careful breeding practices and out crossing - hopefully our salvation.
Page | 4 Kris.Willison_KLM@usa.net 281-804-7801 6-Aug-2008