Ignite Creation Date:
2024-05-06 @ 5:33 PM
Last Modification Date:
2024-10-26 @ 2:30 PM
Study NCT ID:
NCT05340556
Status:
UNKNOWN
Last Update Posted:
2022-04-22
First Post:
2022-04-08
Brief Title:
Microchimerism in Patients With Recurrent Pregnancy Losses
Sponsor:
Caroline Nørgaard-Pedersen
Organization:
Aalborg University Hospital
Study Overview
Official Title:
Microchimerism From a Firstborn Boy or an Older Brother in Patients With Recurrent Pregnancy Losses a Pilot Study
Status:
UNKNOWN
Status Verified Date:
2022-04
Last Known Status:
RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
This pilot study aims to evaluate if microchimeric cells in a patient with recurrent pregnancy loss RPL can be detected by the blood analysis for the presence of the DYS14 gene and the use of indel-panel methods and also to examine if this method can distinguish the cells origin comparing gene sequence from the patients firstborn son or her older brother In addition the pilot study will provide the investigators with information and experience necessary for a subsequent main study to be conducted
Detailed Description:
To the investigators knowledge no previous study has determined the origin of microchimeric cells and therefore this pilot study will use a newly developed genetic analysis which will compare DNA fragments from the male microchimeric cells with DNA fragments from the RPL patients sons and older brothers
The pilot study aims to evaluate the functions and capacity of a newly developed genetic test identifying microchimeric cells A pilot study is necessary to assure the DNA fragments indels analyzed in the genetic analysis include enough informative differences to distinguish between the sons daughters and the older brothers before these investigations can be initiated in a larger sample To assure the test can distinguish origin of microchimeric cells between relatives and work independent of gender The study will include both the probands daughters and sons to strengthen the confidence that the test possesses this ability
10 sRPL and their older brothers and firstborn child is included total included approx 30
Only sRPL whose brothers and children also consent to participate will be included
Blood sample of 12 ml EDTA plasma is collected from the sRPL patient A swap sample from oral mucosa is collected from the older brother and children
The samples will be centrifuged and the buffy coat containing the DNA will be collected from patient and stored at -80 C To detect the multi-copy DYS14 marker located in the TSPY1 gene on the Y-chromosome a real-time PCR analysis will be performed on the extracted DNA with a PCR mastermix specific for the reference gene
Also The extracted DNA will be amplified using specific primers and their associated probes in a multiplex PCR analysis The specific primers target the 10 different indels leading to 10 PCR products with non-overlapping amplicon sizes After the PCR analyses the fragments are analyzed by capillary electrophoresis using GeneticAnalyzer and GeneMapper When differences are identified a qPCR analysis is performed on samples from only the proband with primers and probes specific for the indel fragments that are only present in either the child or the older brother 12 wells loaded with 30000 GE in each well are screened along with a no-template control and a positive control containing 10 GE of DNA homozygous for the allele variant
The pilot study aims to evaluate the functions and capacity of a newly developed genetic test identifying microchimeric cells A pilot study is necessary to assure the DNA fragments indels analyzed in the genetic analysis include enough informative differences to distinguish between the sons daughters and the older brothers before these investigations can be initiated in a larger sample To assure the test can distinguish origin of microchimeric cells between relatives and work independent of gender The study will include both the probands daughters and sons to strengthen the confidence that the test possesses this ability
10 sRPL and their older brothers and firstborn child is included total included approx 30
Only sRPL whose brothers and children also consent to participate will be included
Blood sample of 12 ml EDTA plasma is collected from the sRPL patient A swap sample from oral mucosa is collected from the older brother and children
The samples will be centrifuged and the buffy coat containing the DNA will be collected from patient and stored at -80 C To detect the multi-copy DYS14 marker located in the TSPY1 gene on the Y-chromosome a real-time PCR analysis will be performed on the extracted DNA with a PCR mastermix specific for the reference gene
Also The extracted DNA will be amplified using specific primers and their associated probes in a multiplex PCR analysis The specific primers target the 10 different indels leading to 10 PCR products with non-overlapping amplicon sizes After the PCR analyses the fragments are analyzed by capillary electrophoresis using GeneticAnalyzer and GeneMapper When differences are identified a qPCR analysis is performed on samples from only the proband with primers and probes specific for the indel fragments that are only present in either the child or the older brother 12 wells loaded with 30000 GE in each well are screened along with a no-template control and a positive control containing 10 GE of DNA homozygous for the allele variant
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None