Polymerase chain reaction determination of RhD blood type: an evaluation of accuracy.
OBJECTIVE
To evaluate the accuracy of polymerase chain reaction (PCR) amplification of a portion of the RhD gene by testing a large number of DNA samples derived from individuals whose RhD status was established by the standard serologic method.


METHODS
Seven hundred sixty-five samples were obtained from two sources: subjects taking part in studies at the University of Iowa Hospitals and Clinics (n = 107), and Centre d'Etude du Polymorphisme Humain (CEPH) families used for studies of genetic variation (n = 658). Deoxyribonucleic acid was extracted from blood samples of University of Iowa volunteers and from CEPH families by standard techniques. With few modifications, published primers, reaction and electrophoresis conditions, which yield a 1200-bp fragment in all individuals and a 600-bp fragment in RhD-positive individuals, were used.


RESULTS
By standard serologic techniques, we identified samples from 632 RhD-positive and 133 RhD-negative individuals. Two (both from CEPH) of the 632 RhD-positive individuals were characterized as RhD-negative by PCR. Seven of the 133 RhD-negative samples were judged to be RhD-positive by PCR because of the presence of a light 600-bp band. Despite repeated attempts, no bands or DNA were identified in one RhD-negative sample. Thus, the sensitivity of RhD typing by PCR was 99.7%, the specificity 94.0%.


CONCLUSIONS
Based on our data, it would appear that the use of PCR to establish RhD type can be introduced cautiously into current management schemes in the evaluation of RhD sensitization.
Yankowitz
J
J
Department of Obstetrics and Gynecology, University of Iowa School of Medicine, Iowa City, USA.
Li
S
S
Murray
J C
JC
D001788:Blood Grouping and Crossmatching / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000592:standards*; D011237:Predictive Value of Tests; D011247:Pregnancy; D011296:Prenatal Diagnosis; D015203:Reproducibility of Results; D012203:Rh Isoimmunization / Q000097:blood / Q000175:diagnosis; D012204:Rh-Hr Blood-Group System / Q000235:genetics; D012680:Sensitivity and Specificity
D012204:Rh-Hr Blood-Group System