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  1. #21
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    I think it's normal to lose that many by day 5. It doesn't stop it sucking though. I think it's very unusual to have many make blastie. My sister just did a cycle with good eggs, no apparent problems, 10 mature harvested, 8 fertalized, day 3 I think she still had 8 but come day 5 it was 3.

    My friend is an embryologist and said of my 9 I would probably get 3 to blastie. Obviously some get all of them and some get none. I just hope for something!

    I'm pulling for those other 3 for you though!

  2. #22
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    So aCHG testing... What does it stand for? My clinic always refers to PGD testing which I'm pretty sure is the same thing (or I'm in the wrong thread!!!).

  3. #23
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    April, I know that it is dissappointing but that is a really good result, if you can get another couple tomorrow that is amazing. I had a friend who had 18 embryos and only 3 made it, yet another with 5 that 2 made it. You never know.

    Wise enough, there is getting to blast and then getting to hatching blast that is another total step altogether, I know why my clinic tests on HB but it doesn't make anymore frustrating. aCGH is the type of testing that is done, it is a type of PGD testing. It depends generally on your clnic which type you do. I have used FISH, PCR and now doing this one.

    Of my 6 last time I had 4 blasts, but only 1 suitable for biopsy, most people I know get at least 2. I was told you drop off 50% day 3 to day 5. I have NO idea why my embryos are so suckky, I just wish I had an answer.

    April, like I said in my previous posts I have 3 kids, the 2nd child is a boy and has a lot of health problems, we waited a while to have number 3, because of it. We pretty much started trying straight away and was not getting pregnant, were told to try IVF, so we did, we decided to go overseas and do PGD to get a girl as we totally believe that a girl will be healthier, anyway along the way, we have realised that something is going on with my embryos (3 cycles, 1 no fertilisation, 10 embryos to PGD, 9 abnormal and the one that was considered normal with FISH was a chemical) so consulted with a FS here and FS thinks aCGH is the best way to see if what we transfer is a healthy embryo and also to see if there is actually one thing affecting them all.

    I don't know how I sit with transferring a boy though, on one side my dh and I just want to try and see how it goes but on the other, ds 3 is now showing a lot of the signs of ds2, so if all we get is a healthy boy to transfer, then we may wait until my 2 sons testing is out of the way. We had to wait 6 months to see a specialist (and we had to go interstate to do it, the wait in Perth was 3 years) and testing is still another 6 months away. if they have what they think they have, then I don't think anyone would have a problem with putting only girls in as it is genetic and affects boys 4 times more than girls, if the tests come back clear, then we will be happy to transfer the boy (hypothetically if we can actually get a normal embryo )

  4. #24
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    Quote Originally Posted by Wise Enough View Post
    So aCHG testing... What does it stand for? My clinic always refers to PGD testing which I'm pretty sure is the same thing (or I'm in the wrong thread!!!).
    DNA microarray comparative genomic hybridisation (aCGH) testing is a type of PGD.

    Quote Originally Posted by felicita View Post
    there's an introduction to PGD in this post. (click the link)
    The post referred to above describes Day 3, single cell testing. The problems mentioned with that method are largely resolved by doing Day 5, hatching blast testing where you can collect more cells to start with.
    However, I'm pg with day 5 morulas. Don't know yet if they're chromosomally normal or not (our situation didn't warrant PGD, we did try-it-and-see method instead), if this thread is still going when I find out I'll update. By restricting to day 5 HB, who knows how many good embies are being wasted.
    Every method has it's pros and cons.
    Last edited by felicita; 18-03-2012 at 07:04.

  5. #25
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    My clinic doesn't do hatching blasties, they just drill a hole in the side and take the cells that leak out.

  6. #26
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    April that sucks . Bummer the rest didn't make it. When did they say you would get your biopsy results back? Hopefully all 3 come back as normal.

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    April, I am so so sorry that you didn't get as many as expected but that is a really, really good result. Like previous posters and myself has said getting to the stage your clinic needs is really hard and only the best survive. I am sure you have 1 or 2 normals in there and I hear it all the time, though it is still hard, but you really only need one. I think three is a really good result, I only got 1 with 6 going really well on day 3.

    The success rate of transferring that cgh embryo is really, really high.

    I do tend to wonder though if by having them to get to HB how many good ones are really going to waste, I know tons of people pregnant with day 5 blasts that aren't hatching or day 4 morulas that got pregnant and had healthy babies.

  8. #28
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    April- We also ended up with 3 embryos good enough to biopsy & freeze. I spent the following 2 weeks freaking out that none would be normal & the whole cycle would be futile. I hope you're doing OK while waiting so far and finding some positivity to cling to. There's got to be at least one normal out of your original embie count , maybe even all 3!! The trip to HK sounds sensational, your DS is a very lucky little boy to see Disneyland!

    Wise- the process of drilling the hole is called "assisted hatching" and then the cells emerging through the hole is the beginning of the hatching blastocyst stage. The hyperlink in Felicita's quote didn't work for me, I'll do another post and quote her very user-friendly explanation from our Sydney IVF thread . PGD itself is an umbrella term for any pre-transfer embryo testing. aCGH is a PGD technique that will find issues at the chromosome level. If the condition you're looking to avoid involves a specific gene, then a different PGD technique may be required (eg FISH or PCR). Yay for relocation of your frosties being accomplished! Any news when the nail-biting thaw begins?

    Scarlet- You must be getting close to EPU! How are those follies going? Sending lots of growing vibes . Sorry to hear your DS2 & DS3 have such a long waiting time to get some answers about their health. If your fears are confirmed, then it does sound to me like you'd have justification for gender selection of female embies. Here's hoping there's a perfect princess in the making right now!

    AFM- Trying to stay positive as I know the odds are in my favour this time, but its hard to shake the feeling that if someone has to be in the unlucky 10-20% that get BFN after aCGH, its probably going to be me . So I go from thinking about that one minute to being super-confident the next that this is OUR turn... oh the joys of the TWW!

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  10. #29
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    Inserting the lovely Felicita's post on PGD from another thread:

    Quote Originally Posted by felicita View Post
    Pre-implantation genetic diagnosis.

    Asking if PGD is a common test is like asking if testing your blood sugar is a common test. In the case of blood sugar, if you're diabetic then yes it's common, but if there's no reason to be doing it then it's not. You may enter the PGD world if you (or partner) has a named genetic issue that mustn't be passed on to bubs, or if you've been having repeated problems with your IVF embryo quality or they keep failing to implant.

    PGD is sort of self-explanatory. It is diagnosing genetic issues before you choose to give an embryo the opportunity to implant. If you know you have a genetic issue that mustn't be passed on to bubs then you test all the embryos and only transfer those that pass. They take a cell from a day3 (8cell) embryo for testing. If they get the results quickly you can do a fresh transfer on day 5, otherwise you freeze everything until the results come back and do a FET (frozen embryo transfer) on a later cycle.

    There are limitations to all types of testing. With genetic testing you only get info on the cells you test, so you have to assume that all the cells you didn't test are the same as the one you chose. In an embryo this might not always be true, you might happen to test the only good cell in a bad embryo, or you might test the only bad cell in a good embryo.

    Also, since PGD starts with just 1 cell to test, there's not much DNA to start with, so it might give a false negative result in some tests. If you only look for dodgy genes and it's not there, then that might mean either that all the genes are good, or that that the dodgy bit of DNA got lost somehow before the test was run. A good test will look not only for a dodgy gene, but also for the good version so that you know you've counted the correct number overall.

    There are a lot of different tests that fall under the PGD banner.


    The first is karyotyping.

    karyo = nucleus (part of cell where DNA is kept), with leanings towards implying "chromosomes".

    There are a couple of pictures at http://en.wikipedia.org/wiki/Karyotype .

    Karyotyping is checking that the number and appearance of the chromosomes is OK. Looking at the first of the wikipedia pics (Karyogram of human male using Giemsa staining) you'll see each chromosome is stripey (zoom in if necessary). Part of the test uses Giemsa dye to stain the chromosomes, and each one has its own characteristic pattern. Based on the pattern and the size/shape of the chromosome they can be arranged into their matching 23 pairs.

    tri = 3
    soma = body
    trisomic = 3 objects

    Trisomic conditions, like Downs, will be spotted because there'll be an extra chromosome (#21 for Downs). Adults can also be karyotyped. All you need is a cell with a nucleus, so it's done as a blood test (using white blood cells). Many of the girls (and partners) here will have had the test. It's a first, general look at things.

    Because of the banding/stripey patterns you can also spot if two chromosomes have swapped tails with each other (using the colourful method of the last picture might make swaps more obvious). This is a translocation. (trans = across.) If you do the swap neatly and don't lose any DNA then it is a balanced translocation. You still have all the correct info, but it's not in the right place. There's usually no problem with your own health with a balanced translocation, but it makes problems for TTC.

    (If it helps, print the either the first or the last (Spectral karyogram of a human female) pic from wikipedia and get a pair of scissors and sticky tape.) Swap over half of one chromosome#1 with half of one chromosome #2 to make a balanced translocation. Everything is still fine in that person, because they've got all the info. But when they go to make eggs or sperm, they only want to pass on 1 of each chromosome. When doing the lucky dip to decide which of each pair to pass on everything will be fine if they happen to grab the chromosomes you didn't swap anything on. One copy of everything will be passed on as per requirements.

    If they grab both of the translocated chromosomes then they also pass on all the required DNA, just that half of chromosome#2's DNA is located on #1, and vice versa.
    But if they grab the half-and-half #1 and the intact #2 then they'll pass on twice as much DNA for part of #2, and be missing half of the #1 DNA. This is a problem for bubs. In some cases (depending on exactly which genes are involved) it might only be a little problem, but usually it's a big problem.



    A little step up from karyotyping is FISH.
    Fluorescence in situ hybridisation.

    Not sure of your base knowledge of DNA, so forgive me if this is too simple. The four letters used in the DNA alphabet are GATC. These 4 letters refer to the first letter of the name of the chemical structure in the DNA.

    DNA is often described as being like a ladder. The ladder comes in a left and right half, and in order to stick it together properly the rungs have to match up, so that a G on one side matches a C on the other, and an A has to match a T. If you try to match a G with an A the sides of the ladder are going to be bulged outwards, and the glue in the middle won't be sticky. If you try to match a C with a T the rungs won't meet in the middle. G+T, or A+C will be the right length, but no glue.

    (I remember the pairs visually. G and C are the round letters so they match. A and T are the straight letters so they match.)

    If I happen to know that a particular sequence of DNA reads GAATTCA, then I can make my own little sequence that matches it (CTTAAGT, wherever there was a G I wrote a C, etc.) and label my little piece with a fluorescent tag. My piece can now be a useful tool called a "probe". Going back to the karyotyped DNA, I can heat that DNA up which weakens the ladder glue, then throw my probe into the mix and let it all cool down. My little bit should insert itself into the ladder wherever it finds its match (as an irrelevant side effect, the proper long side of the ladder is forced to detour around my probe.) Since my probe fluoresces, then when I look at the chromosomes I should see a couple of bright dots wherever my probe has stuck.
    http://en.wikipedia.org/wiki/Fluores..._hybridization
    The first wikipedia picture has a green probe that matches one sequence of DNA, and a red probe that matches a different DNA sequence. That picture isn't from a normal cell, so that's all I'm going to say about it.

    Now back to PGD. If you know you have a particular gene that's causing a problem then you can make a probe for it and use FISH to see if it's present in your test sample. FISH gives you info on not only what gene is present, but also on how many copies of the gene there are and where it is found.


    PCR is a little bit similar to FISH.
    Polymerase chain reaction.

    PCR is a way of increasing the amount of DNA you have to work on in the lab (using a chain reaction to make a polymer of DNA).
    In order to kick off the PCR you need small pieces of DNA like the FISH probes, but without the fluorescent label attached. For PCR, you use 2 probes, and the chain reaction increases the amount of chromosome DNA that's located between where those probes attach.

    If you just want to increase the amount of all DNA present then you use probes with random sequences to hopefully include everything in your net. If you're only interested in one gene, then you choose your probes carefully using the known gene DNA sequence so that you only increase the small amount of DNA that you're interested in.

    When used diagnostically PCR gives you information on whether the DNA you're looking for is present, but tells you nothing about which chromosome it's on. And when the DNA your looking for is present, PCR doesn't usually say if you have 1 or 2 (or more) copies of it. It's more of a presence/absence (qualitative) test, not a quantitative test. But it's easier to do than FISH. For both PCR and FISH you have to have an idea of what you're looking for before you do the test, because you have to make your probes in advance.

    (For completeness, the probes used to kick off PCR are called primers.)


    Now there's a "new" test, CGH, which, like karyotyping, can be used when you don't know what problem you're looking for.
    Comparative genomic hybridisation.
    (I've ripped this explanation from http://www.bubhub.com.au/community/f...r-Women/page48 )

    The principle behind CGH is you get:

    (a) a sample of "proper" reference DNA and label it a fluorescent colour like green.
    (b) a sample of the emby's DNA and label is a different fluorescent colour like red.

    Heat an equal amount of both samples up together in the same container so that the rungs of the DNA ladder separate down the middle like a zipper. This leaves the labelled DNA looking for its match when you let it cool down.

    Apply the unzipped DNA to a test chip which has unlabelled fragments of DNA already stuck to it so that the fluorescent labelled bits have specific places to zip themselves up to. This forms lots of fluorescent spots on the test chip. A lot of work would have gone into identifying which spots come from which bit of chromosome, and to making sure that each bit of chromosome only had an affinity for one particular spot, before the technique was released for general use.

    Use a computer to read the colour of each of the spots. If the colour is yellow then there's an equal amount of DNA (a) and (b) so the emby has the right number of that bit of DNA. (Fluorescent red + fluorescent green = fluorescent yellow)
    If there's more green then the emby is missing a bit, so more of the reference sample stuck to the spot.
    If there's more red then the emby had extra DNA for that bit, so more of the emby's DNA stuck to that spot.

    There's a bit of random wiggle in the colour of the results, but if, say, all the spots for chromosome 4 are red then the emby probably had a trisomy for that. It will also spot unbalanced translocations, and smaller regions of extra/absent DNA where particular spots on the test chip show up too much or too little red or green.


    There's a picture of coloured dots at http://www.brookscole.com/chemistry_...GeneChips.html . The brightness of the dot is due to the amount of DNA/label present. The brightness is not as important as colour (red - yellow - green).

    Hope that helps.

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  12. #30
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    Lindylou, I know the wait must be hard, are you temped to do a home pregnancy test? I know when I had my transfer I couldn't stop myself.

    I actually do know someone who had a aCGH embryo transferred and got a BFN, I am not saying that to worry you, but since they knew it was a chromosomally normal embryo they went on to do specific immune testing and have found that this may be her problem, so although it was horrible for her to go through it, she does now have a reason for it and is working on the immune issues before her next transfer.

    With me, I had a scan yesterday and had 6 over 13mm and 3 between 10-13, my clinic only counts the 13mm ones at this stage. I have to do another 2 days stims and have a scan tomorrow and hopefully trigger tomorrow night, EPU on Friday. My estrogen was 2600.

    This has been a strange cycle, I have only just started feeling anything in my ovaries and now just getting the sick feeling I get when my estrogen gets higher, I am also getting the tiredness. I am also doing stims for at least 12 days, I really hope it will be all right. I don't think I will ever do another back to back cycle again.

    I am booking today to fly to Sydney tomorrow, just hoping for at least another one for biopsy.


 

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