Celomic Fluid: Laboratory Workflow for Prenatal Diagnosis of Monogenic Diseases.


Journal

Molecular diagnosis & therapy
ISSN: 1179-2000
Titre abrégé: Mol Diagn Ther
Pays: New Zealand
ID NLM: 101264260

Informations de publication

Date de publication:
03 2022
Historique:
accepted: 04 01 2022
pubmed: 18 2 2022
medline: 23 4 2022
entrez: 17 2 2022
Statut: ppublish

Résumé

Celomic fluid can be considered as an ultra-filtrate of maternal serum, containing a high protein concentration, urea, and many other molecules. It is an important transfer interface and a reservoir of nutrients for the embryo. Celomic fluid contains fetal cells that can be used for prenatal diagnosis of monogenic diseases in an earlier gestational period than villocentesis and amniocentesis. The purpose of this study was to evaluate the characteristics of celomic fluid and to establish a workflow laboratory procedure for very early prenatal diagnosis of monogenic diseases. Three hundred and eighty-five celomatic fluids were collected between the seventh and tenth week of gestation. We sampled 1 mL of celomic fluid in all cases. The embryo-fetal erythroid precursor cells were selected by the anti-CD71 microbead method or by a direct micromanipulator pick-up on the basis of their morphology. We amplified the extracted DNA using a nested polymerase chain reaction. Primers for short tandem repeat amplification were used to perform a quantitative fluorescent polymerase chain reaction evaluation to control maternal contamination. We observed maternal contamination in 95% of celomic fluids with a range between 5 and 100%. No fetal cells were observed in 0.78% of celomic fluids. The number of fetal cells ranged from a few units to several hundred. Isolation of embryo-fetal erythroblasts selected by the micromanipulator made diagnosis feasible in all cases. The selection of fetal cells by a micromanipulator and nested polymerase chain reaction analysis made celomatic fluid suitable for early prenatal diagnosis of monogenic disorders even in the presence of high maternal contamination and few fetal cells. The procedure reported in this study provides the opportunity for the use of celomic fluid sampled by celocentesis as an alternative to chorionic villi sampling and amniocentesis, to allow invasive prenatal diagnosis at a very early stage of pregnancy.

Sections du résumé

BACKGROUND
Celomic fluid can be considered as an ultra-filtrate of maternal serum, containing a high protein concentration, urea, and many other molecules. It is an important transfer interface and a reservoir of nutrients for the embryo. Celomic fluid contains fetal cells that can be used for prenatal diagnosis of monogenic diseases in an earlier gestational period than villocentesis and amniocentesis.
OBJECTIVE
The purpose of this study was to evaluate the characteristics of celomic fluid and to establish a workflow laboratory procedure for very early prenatal diagnosis of monogenic diseases.
METHODS
Three hundred and eighty-five celomatic fluids were collected between the seventh and tenth week of gestation. We sampled 1 mL of celomic fluid in all cases. The embryo-fetal erythroid precursor cells were selected by the anti-CD71 microbead method or by a direct micromanipulator pick-up on the basis of their morphology. We amplified the extracted DNA using a nested polymerase chain reaction. Primers for short tandem repeat amplification were used to perform a quantitative fluorescent polymerase chain reaction evaluation to control maternal contamination.
RESULTS
We observed maternal contamination in 95% of celomic fluids with a range between 5 and 100%. No fetal cells were observed in 0.78% of celomic fluids. The number of fetal cells ranged from a few units to several hundred. Isolation of embryo-fetal erythroblasts selected by the micromanipulator made diagnosis feasible in all cases.
CONCLUSIONS
The selection of fetal cells by a micromanipulator and nested polymerase chain reaction analysis made celomatic fluid suitable for early prenatal diagnosis of monogenic disorders even in the presence of high maternal contamination and few fetal cells. The procedure reported in this study provides the opportunity for the use of celomic fluid sampled by celocentesis as an alternative to chorionic villi sampling and amniocentesis, to allow invasive prenatal diagnosis at a very early stage of pregnancy.

Identifiants

pubmed: 35175567
doi: 10.1007/s40291-022-00577-3
pii: 10.1007/s40291-022-00577-3
doi:

Substances chimiques

DNA 9007-49-2

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

239-252

Informations de copyright

© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.

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Auteurs

Antonino Giambona (A)

Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis, and Rare Hematological Diseases, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Via Trabucco 180, 90146, Palermo, Italy. a.giambona@villasofia.it.

Margherita Vinciguerra (M)

Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis, and Rare Hematological Diseases, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Via Trabucco 180, 90146, Palermo, Italy.

Filippo Leto (F)

Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis, and Rare Hematological Diseases, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Via Trabucco 180, 90146, Palermo, Italy.

Filippo Cassarà (F)

Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis, and Rare Hematological Diseases, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Via Trabucco 180, 90146, Palermo, Italy.

Viviana Tartaglia (V)

Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis, and Rare Hematological Diseases, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Via Trabucco 180, 90146, Palermo, Italy.

Valentina Cigna (V)

Unit of Fetal Medicine and Prenatal Diagnosis, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Palermo, Italy.

Emanuela Orlandi (E)

Unit of Fetal Medicine and Prenatal Diagnosis, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Palermo, Italy.

Francesco Picciotto (F)

Unit of Fetal Medicine and Prenatal Diagnosis, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Palermo, Italy.

Nourah H Al Qahtani (NH)

Obstetrics and Gynecology Department, College of Medicine, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

Eman S Alsulmi (ES)

Obstetrics and Gynecology Department, College of Medicine, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

Noor B Almandil (NB)

Department of Clinical Pharmacy Research, Institute for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

Sayed AbdulAzeez (S)

Department of Genetic Research, Institute for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

J Francis Borgio (JF)

Department of Genetic Research, Institute for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

Aurelio Maggio (A)

Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis, and Rare Hematological Diseases, Azienda Ospedaliera, Ospedali Riuniti Villa Sofia Cervello, Via Trabucco 180, 90146, Palermo, Italy.

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