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== Extraction of test samples ==
[[Blood plasma]] is commonly used as test samples for verifying the maternal RhD status. Blood plasma can also be used for determining the foetal RhD status if the mother is RhD- as maternal blood plasma contains maternal [[DNA]] and trace amounts of foetal DNA.<ref name="Reliable Determination of Fetal RhD">{{cite journal | vauthors = Dovč-Drnovšek T, Klemenc P, Toplak N, Blejec T, Bricl I, Rožman P | title = Reliable Determination of Fetal RhD Status by RHD Genotyping from Maternal Plasma | journal = Transfusion Medicine and Hemotherapy | volume = 40 | issue = 1 | pages = 37–43 | date = February 2013 | pmid = 23637648 | pmc = 3636019 | doi = 10.1159/000345682 }}</ref> Blood samples can be obtained through [[venipuncture]] of the mother. Since plasma and other components of blood have different densities, [[centrifugation]] of blood samples with added [[anticoagulant]] (such as [[Ethylenediaminetetraacetic acid|EDTA]]) can segregate blood contents into multiples layers.<ref>{{cite journal | vauthors = Dagur PK, McCoy JP | title = Collection, Storage, and Preparation of Human Blood Cells | journal = Current Protocols in Cytometry | volume = 73 | pages = 5.1.1–16 | date = July 2015 | pmid = 26132177 | pmc = 4524540 | doi = 10.1002/0471142956.cy0501s73 | isbn = 9780471142959 }}</ref> Blood plasma can then be isolated from the other components for rhesus factor testing. The method of extracting foetal DNA from maternal blood plasma is considered to be a type of non-invasive [[prenatal testing]].<ref>{{Cite web|url=https://www.bbts.org.uk/blog/noninvasive_prenatal_testing_for_fetal_rhesus-d_status_-_putting_the_/|title=BBTS {{!}} Non-invasive prenatal testing for fetal rhesus-D status - putting the {{!}}|website=www.bbts.org.uk|access-date=2019-04-09}}</ref>
▲=== Non-invasive extraction ===
Non-invasive [[prenatal testing]] can be used if the mother is RhD-.<ref>{{cite journal | vauthors = Saramago P, Yang H, Llewellyn A, Walker R, Harden M, Palmer S, Griffin S, Simmonds M | title = High-throughput non-invasive prenatal testing for fetal rhesus D status in RhD-negative women not known to be sensitised to the RhD antigen: a systematic review and economic evaluation | journal = Health Technology Assessment | volume = 22 | issue = 13 | pages = 1–172 | date = March 2018 | pmid = 29580376 | pmc = 5890172 | doi = 10.3310/hta22130 }}</ref> However, in the case of maternal RhD status being negative, invasive prenatal testing may be used to determine the foetal RhD status instead. The two most common invasive methods of extracting foetal DNA are [[chorionic villus sampling]] (CVS) and [[amniocentesis]] (AMC).<ref>{{cite journal | vauthors = Carlson LM, Vora NL | title = Prenatal Diagnosis: Screening and Diagnostic Tools | journal = Obstetrics and Gynecology Clinics of North America | volume = 44 | issue = 2 | pages = 245–256 | date = June 2017 | pmid = 28499534 | pmc = 5548328 | doi = 10.1016/j.ogc.2017.02.004 }}</ref> These invasive procedures can be conducted on both RhD+ and RhD- mothers. After the invasive procedure, medications that prevent the Rh [[Immunization|immunisation]] are usually prescribed to RhD- mothers<ref>{{Cite journal|last=Crowther|first=Caroline A.|last2=Middleton|first2=Philippa|last3=McBain|first3=Rosemary D.|editor1-first=Caroline A|editor1-last=Crowther|date=2013-02-28|title=Anti-D administration in pregnancy for preventing Rhesus alloimmunisation|journal=The Cochrane Database of Systematic Reviews|issue=2|pages=CD000020|doi=10.1002/14651858.CD000020.pub2|issn=1469-493X|pmid=23450526}}</ref>. This is done to avoid the production of maternal anti-D [[Antibody|antibodies]] which may attack the foetal blood cells should the foetus be Rh incompatible with the mother.<ref name="Mechanisms of anti-D action in the">{{cite journal | vauthors = Brinc D, Lazarus AH | title = Mechanisms of anti-D action in the prevention of hemolytic disease of the fetus and newborn | journal = Hematology. American Society of Hematology. Education Program | volume = 2009 | pages = 185–91 | date = 2009 | pmid = 20008198 | doi = 10.1182/asheducation-2009.1.185 | url = https://semanticscholar.org/paper/d43570abd49c16a6e563c9a379074ff4b5024ec6 | doi-access = free }}</ref>▼
=== Invasive extraction ===
▲Non-invasive [[prenatal testing]] can be used if the mother is RhD-.<ref>{{cite journal | vauthors = Saramago P, Yang H, Llewellyn A, Walker R, Harden M, Palmer S, Griffin S, Simmonds M | title = High-throughput non-invasive prenatal testing for fetal rhesus D status in RhD-negative women not known to be sensitised to the RhD antigen: a systematic review and economic evaluation | journal = Health Technology Assessment | volume = 22 | issue = 13 | pages = 1–172 | date = March 2018 | pmid = 29580376 | pmc = 5890172 | doi = 10.3310/hta22130 }}</ref> However, in the case of maternal RhD status being negative, invasive prenatal testing may be used to determine the foetal RhD status instead. The two most common invasive methods of extracting foetal DNA are [[chorionic villus sampling]] (CVS) and [[amniocentesis]] (AMC).<ref>{{cite journal | vauthors = Carlson LM, Vora NL | title = Prenatal Diagnosis: Screening and Diagnostic Tools | journal = Obstetrics and Gynecology Clinics of North America | volume = 44 | issue = 2 | pages = 245–256 | date = June 2017 | pmid = 28499534 | pmc = 5548328 | doi = 10.1016/j.ogc.2017.02.004 }}</ref> These invasive procedures can be conducted on both RhD+ and RhD- mothers. After the invasive procedure, medications that prevent the Rh [[Immunization|immunisation]] are usually prescribed to RhD- mothers<ref>{{Cite journal|last=Crowther|first=Caroline A.|last2=Middleton|first2=Philippa|last3=McBain|first3=Rosemary D.|editor1-first=Caroline A|editor1-last=Crowther|date=2013-02-28|title=Anti-D administration in pregnancy for preventing Rhesus alloimmunisation|journal=The Cochrane Database of Systematic Reviews|issue=2|pages=CD000020|doi=10.1002/14651858.CD000020.pub2|issn=1469-493X|pmid=23450526}}</ref>. This is done to avoid the production of maternal anti-D [[Antibody|antibodies]] which may attack the foetal blood cells should the foetus be Rh incompatible with the mother.<ref name="Mechanisms of anti-D action in the">{{cite journal | vauthors = Brinc D, Lazarus AH | title = Mechanisms of anti-D action in the prevention of hemolytic disease of the fetus and newborn | journal = Hematology. American Society of Hematology. Education Program | volume = 2009 | pages = 185–91 | date = 2009 | pmid = 20008198 | doi = 10.1182/asheducation-2009.1.185 | url = https://semanticscholar.org/paper/d43570abd49c16a6e563c9a379074ff4b5024ec6 | doi-access = free }}</ref>
==== Chorionic villus sampling ====
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