First successful pregnancy after preimplantation genetic diagnosis for Hemophilia A in Asia
8/08/2013 12:32:00 AM
First successful pregnancy after preimplantation genetic diagnosis for Haemophilia A in Asia
| Author | Hsing-Hua Lai, Huai-Ling Wang, Shih-Chieh Huang, Wen-Yi Chiang |
|---|---|
| Editor | Hui-Ling Kao |
| Seminar name | 2010 TSRM |
| Keyword | PGD, inherited disease, IVF |
| PDF download |
Haemophilia A, the most common X-linked disorder affecting males, results from deficient or defective coagulation factor VIII which impairs the coagulation cascade (Fig. 1) and causes bleeding symptoms. In Taiwan, there are about 1000 haemophilia patients, where the cost on medication is 2,300 million dollars per year. Different types of factor VIII gene mutation have been characterized and among which, intron 22 inversion is the most seen and responsible for 45% of severe HA cases.
The conventional way for HA patients to have an unaffected child is to play “reproductive roulette” and give up all male babies. Preimplantation genetic diagnosis (PGD) provides an alternative strategy for couples carrying HA to conceive a healthy child without sex selection hence to reduce the mental pressure and medical cost. This technique can be performed on blastomere or trophectoderm cells. The DNA in each cell is analyzed for a specific gene mutation, i.e. factor VIII gene in HA. In recent years, the need of PGD for HA is increasing.
Material and Method
Embryo culture, biopsy and cryopreservation
All embryos were cultured in single media (LifeGlobal). On day 3 post-oocyte retrieval, embryos that were in ≥6 cell stage and had ≤20% fragmentation were biopsied with non-contact laser. From each embryo, one blastomere was aspirated, collected separately into a PCR tube, stored at -20℃ and then sent to a standard PGD laboratory for factor VIII gene intron 22 inversion analysis.
Embryos were further cultured to day 5 and vitrified by Cryotop method. After the PGD results came out, frozen/thawed unaffected embryos with best quality were transferred. Thawing procedure was as described in Cryotop method.
Mutation analysis and PGD
Patient buccal cell DNA was obtained for construction of customized DNA probe. DNA from each blastomere was tested by a polymerase chain reaction (PCR) protocol using ABI 3130 Genetic Analyzer. Factor VIII gene amplification was followed by the second PCR round using the customized probes as primers. The final PCR products were subjected to capillary electrophoresis and analyzed by GeneMapper for allele identification.
Results
Patient
The patient is a 38-year-old HA carrier female whose first son died from HA in infant age. Family linkage analysis was performed earlier and the mutation type was determined as intron 22 inversion. 10 years after, she came to our clinic for the help of IVF because of diminished ovarian reserve. Pregnancy was achieved but ended in termination due to HA affected fetus. Later on, she approached PGD and went through IVF again.
IVF results
In total, ten mature oocytes were retrieved and nine zygotes were obtained. At day 3, all nine embryos were qualified to biopsy. Two embryos stopped growing at day 4. Till day 5, embryo vitrification was carried out on the other seven embryos.
PGD results
The factor VIII genotype was determined based on amplicon size compared with the maternal microsatellite markers. The data showed in Fig. 2 were two different markers and two representative embryo allele status (normal and affected). Nine specimens were successfully analyzed and the PGD results were: 1 carrier, 4 mutant and 4 normal embryos (Tab. 1). The genotyping rate was 100%.
Tab. 1 PGD results.
| No. | 1 | 2 | 3 | 4 | 5 |
|---|---|---|---|---|---|
| Embryo stage (D5) | Early blastocyst | Arrested | Early blastocyst | Early blastocyst | Early blastocyst |
| Genotype | carrier | mutant | normal | normal | mutant |
| No. | 6 | 7 | 8 | 9 |
|---|---|---|---|---|
| Embryo stage (D5) | Blastocyst | Blastocyst | Early blastocyst | Arrested |
| Genotype | mutant | normal | normal | mutant |
Clinical outcome
Single frozen/thawed blastocyst with normal factor VIII genotype (embryo 7 in Tab. 1) was transferred back on day 21 of the patient’s natural cycle. Pregnancy was achieved. Serum b-hCG was 2004.43 mIU/mL on day 16 after embryo transfer. Amniocentesis was concordant with the PGD result as a normal male baby.
Conclusion
We report a successful pregnancy of a HA carrier with the help of PGD, proving that PGD as an alternative diagnosis in HA is feasible.
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kindly email DR EHINOMEN for your own cure if you have any disease listed above via drehinomenherbalhome@gmail.com
Do you need a financial help? Are you in any financial crisis or do you need funds to start up your own business? Do you need funds to settle your debt or pay off your bills or start a good business? Do you have a low credit score and you are finding it hard to obtain capital services from local banks and other financial institutes? Here is your chance to obtain a financial services from our company. We offer the following finance to individuals-
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Do you need a financial help? Are you in any financial crisis or do you need funds to start up your own business? Do you need funds to settle your debt or pay off your bills or start a good business? Do you have a low credit score and you are finding it hard to obtain capital services from local banks and other financial institutes? Here is your chance to obtain a financial services from our company. We offer the following finance to individuals-
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Mrs.Carol AndersonDo you need a financial help? Are you in any financial crisis or do you need funds to start up your own business? Do you need funds to settle your debt or pay off your bills or start a good business? Do you have a low credit score and you are finding it hard to obtain capital services from local banks and other financial institutes? Here is your chance to obtain a financial services from our company. We offer the following finance to individuals-
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HOW I GOT CURED OF HERPES DISEASE
ReplyDeleteI\'m here to testify about what DR. OLU did for me. I have been suffering from (HERPES) disease for the past 5 years and had constant pain, especially in my knees. During the first year,I had faith in God that i would be healed someday.This disease started circulating all over my body and i have been taking treatment from my doctor, few weeks ago i came across a testimony of one lady on the internet testifying about a Man called DR.OLU on how he cured her from herpes disease. And she also gave the email address of this man and advise anybody to contact him for help for any kind of sickness that he would be of help, so I emailed him telling him about my (HERPES) he told me not to worry that i was going to be cured!! Well i never believed it,, well after all the procedures and remedy given to me by this man few weeks later i started experiencing changes all over me as DR. OLU assured me that i will be cured,after some time i went to my doctor to confirmed if i have be finally healed behold it was TRUE, the test came out negative. So friends my advise is if you have such sickness or any other at all you can contact DR. OLU for help via email} drolusolutionhomehome@gmail.com/ Or CALL/WHATAPPS him through his mobile} +2347014784614
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ReplyDeleteThanks to Dr Williams I am so happy today, my wife have been suffering from hemophilia for the past 8 years now, and i have spent a lot on western drugs which has all proved abortive, i have tried all means in life to get my wife out of this , but there was no answer until i decided to try herbal solution and i found Dr Williams online and i contacted him and after she took his medication as instructed, my wife is now completely cure from hemophilia within those week of usage, i am so much happy, thanks to Dr Williams for helping me get my wife life back again without any form of crisis, i promise to tell your name and good deeds to the whole world,if you have someone with hemophilia you can email him on drwilliams098675@gmail.com for more information .
ReplyDelete
ReplyDeleteHappiness is all i see now I never thought that I will be cured from HERPES virus again. I have been suffering from a deadly disease (HERPES) for the past 3 years now, I had spent a lot of money going from one places to another, from churches to churches, hospitals have been my home every day residence. Constant checks up have been my hobby not until this faithful day, I was searching through the internet, I saw a testimony on how Dr Ogudugu helped someone in curing his HERPES disease, quickly I copied his email which is greatogudugu@gmail.com just to give him a test I spoke to him, he asked me to do some certain things which I did, he told me that he is going to provide the herbal cure to me, which he did, then he asked me to go for medical checkup after some days after using the herbal cure, behold I was free from the deadly disease, he only asked me to post the testimony through the whole world, faithfully am doing it now, please brothers and sisters, he is great, I owe him in return. If you are having a similar problem just email him on ( greatogudugu@gmail.com ) or you can whatsApp his mobile number on +27663492930
i never believed that their could be any complete cure for fibroid with out going for surgery ,i saw people’s testimony on blog sites and Facebook page of how Dr Omohan prepare herbal cure for different king of disease,and with the help of his herb they where able to shrink there fibroid naturally. i had to try it too and you can,t believe that in just few weeks i started using it all my pains stop gradually . Right now i want to tell you all that i just give birth to a baby boy last month,and on thin now i have not had any pain, and i have just went for text last week and the doctor confirmed that there is no trace of any fibroid in my system . Glory be to God for leading me to this great Dr omohan I am so happy as i am sharing this testimony. My advice to you all who thinks that their is no herbal cure for fibroid that is Not true ,just contact him and get cure throughhis email: dromohanherbalmedicine@gmail.com or whatsapp +2348164816038
ReplyDeleteCow/ Ox Gallstones available
ReplyDeleteSpecifications
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I am testifying about a great herbal doctor who saved me from AZOOSPERMIA , his name is Dr. Uduehi. I suffered zero sperm for many years, I’ve married twice and no woman could stay because of my inability to make them pregnant. I read about Uduehi’s medication online and I decided to try my luck, I contacted him and after using his medication my sperm count was 8 thousand and just like that it keeps increasing and its above 16 million as I speak and my wife is pregnant You too can become fertile just like me if interested. Contact him through: uduehiherbalcare@gmail.com phone/WhatsaPP: +2347084878384
ReplyDeleteI'm Catherine and i am here to share a great story of how my 11 years old son was cured of an autistic disease, I have gone to so many places but to no avail, i have spent a lot of money from hospitals trying to eradicate this disease and when all hope was lost, I was introduced by a friend to Dr Odoin at (dr.odionsolutionhome@yahoo.com) who helped me cure Jones my son from his Autistic disease. Medical doctors could only give me supplements to suppress this disease but Jones will always exhibit this symptoms of autism; difficulty with communication, difficulty with social interactions, obsessive interests and repetitive behaviors and non verbal but Dr Odion cured my son Jones totally from this autistic symptoms. This is so great because Jones is my only son and i couldn't bear the social disgrace and embarrassment Jones always experienced. But thanks a lot to this great doctor who helped me in so many ways to cure my son Jones who is now a normal and healthy kid. It's been 2 months since i encountered with this great doctor and since then Jones has never portrayed any of this symptoms anymore nor undergo crisis and now plays with his friends. Though he experiences some little stigmatization about his past condition but he is a 100 percent normal kid now all thanks to this great doctor. If you're in need of cure for this autistic disease you can contact the doctor who cured Jones at (dr.odionsolutionhome@yahoo.com). All medical reports now shows that Jones is free from Autism. I write this here today because i am so glad that my son is healed and i would also want you to be cured if faced with this disease, indeed i am grateful to Dr Odion for curing Jones.
ReplyDeleteHemophilia is a rare disorder in which your blood doesn't clot normally because it lacks sufficient blood-clotting
ReplyDeleteproteins (clotting factors). If you have hemophilia,you may bleed for a longer time after an injury than you would if your
blood clotted normally.
What a beautiful and wonderful testimony, sometimes things you don't believe can just happen. My name is Adams Vera, from U.S.A am 28 years old i got married at the age of 24 i have only one child and i was living happily. After three years of my marriage my husband behavior became so strange and i don't really understand what was going on, he packed out of the house to another woman i love him so much that i never dreamed of losing him, i try my possible best to make sure that my husband get back to me but all to no avail i cry and i cry seeking for help because i was heart broken until i coincidentally came across an online spell caster called Dr. Osasu , i contacted him and explained to him what i have been going through in my marriage and he gave me the full assurance that my husband will come back to me and my marriage will be restored, He told me what to do to get my husband back and i did, he said after 3days my husband will come back to me and start begging, it really happen it was so surprising and everything was just like a miracle, ever since my family have been living with so much happiness and our relationship was now very tight because of this great and powerful spell caster.he is a very great and powerful spell caster that can be trusted and there is nothing concerning life issues he cannot solve because i am a living testimony. i know there are so many people out there who are going through similar problems in one way or the other, believe me this is the right spell caster to contact and all your problems will be forgotten. contact Dr. Osasu viaWhatsapp: +2347064365391 Email:Account drosasu25@gmail.com
ReplyDeleteHemophilia Clinic. The Hemophilia
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bleeding disorders
I never thought i would be HIV negative again after been diagnosed in 2017, i have tried everything possible in life from one doctor to another, one hospital to another, series of tests, different kinds of medication, i had already lost hope until i meet Great Dr. OSAGIE online testimonies, a specialist in herbal medication from Africa, i contacted him (drosagiesolutiontemple@gmail.com OR
ReplyDeleteDROSAGIESOULTIONTEMPLE@YAHOO.COM) and he prepared HIV herbal medication for me which i took for 7days and now i am completely cured. i want to use this medium to express my gratitude to him for saving my life and curing me from HIV, for taking away all my pains and sorrows, I''m indeed grateful and i am so happy I''m now HIV negative. i will continue to tell the good news of your great works to everyone, if you have HIV or other disease contact him, Email: his email: (drosagiesolutiontemple@gmail.com OR
DROSAGIESOULTIONTEMPLE@YAHOO.COM ) or Whatsapp number: +2347030465649
DOCTOR OSAGIE CAN AS WELL HELP THE FOLLOWING PROBLEMS
1. HIV/AIDS SPELL
2. HERPES CURE OF KIND
3. CANCER SPELL
4 IF YOU WANT YOUR EX LOVER BACK SPELL
5 IF YOU NEED A BABY SPELL him to solve
6 LOW SPERM COUNT SPELL
get all your problem solve. No problem is too big for him to solve.
Finally I'm free from AZOOSPERMIA after years of no sperm in my testicles, but I never take it seriously none did I see it as a problem while growing up until I became of age to settle down with a woman. My woman advised me to go hospital for diagnoses that its abnormal as she has not felt my sperm in her ever since we started dating, I became very worried and confused when I was told at the hospital that I cannot impregnate a woman. because there's no sperm, I went looking for solution but all to no avail, until a came across Dr. Uduehi’s recommendation online. I reached out to him and for the first time I start to experience sperm after taking his medication, it was little at first but became plenty later. You can reach him via Tel: +234-708-487-8384 Email: uduehiherbalcare@gmail.com
ReplyDeleteFinally I'm free from AZOOSPERMIA after years of no sperm in my testicles, but I never take it seriously none did I see it as a problem while growing up until I became of age to settle down with a woman. My woman advised me to go hospital for diagnoses that its abnormal as she has not felt my sperm in her ever since we started dating, I became very worried and confused when I was told at the hospital that I cannot impregnate a woman. because there's no sperm, I went looking for solution but all to no avail, until a came across Dr. Uduehi’s recommendation online. I reached out to him and for the first time I start to experience sperm after taking his medication, it was little at first but became plenty later. You can reach him via Tel: +234-708-487-8384 Email: uduehiherbalcare@gmail.com
ReplyDeleteI did surgery for multiple FIBROID 2 years ego and they grow back after sometime with all the pains included I couldn't get pregnancy also. I told a friend about it because I was confused, and she told me her own experience about fibroid and how she got raid of it through Dr. Uduehi. I contacted the doctor and the fibroid started shrinking down even before I completed the medication. I couldn’t testify of it until now that am 4 months pregnant, I also went to the hospital and was diagnosed negative and fibroid free. Contact the doctor through Email: uduehiherbalcare@gmail.com or +234-708-487-8384
ReplyDelete