PubMed · 6810321
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E Cundy. 1982-08-04. Cell separation: a positive process.. https://pubmed.ncbi.nlm.nih.gov/6810321/
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The objective of this study was to evaluate the accuracy of a DNA-based testing methodology in determining the RhD genotypes of fetuses at risk for RhD hemolytic disease. We designed a multiplex polymerase chain reaction-based test based on recent RhD and RhCE sequence information. To improve the accuracy of the results, two different portions of the RhD gene were examined. Deoxyribonucleic acid was extracted from fetal specimens, portions of the RhD gene were amplified by the polymerase chain reaction, and the amplified product was run on a polyacrylamide gel to look for the presence or absence of the RhD gene. We tested 67 amniotic fluid and two chorionic villus specimens to determine the fetal RhD genotype in pregnancies at risk for RhD hemolytic disease. Forty-seven of the 69 specimens were determined to be Rh-positive, and 22 were Rh-negative. Fifty of the 69 fetal specimens--31 Rh-positive and 19 Rh-negative--were serotyped at birth. In all 50, there was complete correlation between the DNA analysis and the serotyping results. RhD gene analysis is a rapid and reliable method that provides an accurate fetal genotype to aid in the prenatal care of RhD-alloimmunized women.
We describe the clinical and biochemical features of an infant with marked transient porphyrinemia in whom blistering developed while the infant was undergoing phototherapy for severe Rh isoimmunization. The cause of the transient porphyrinemia was likely to be multifactorial--abnormal porphyrin metabolism or accumulation in a premature infant with multisystem disease and multiple drug therapy. In addition, the infant received an unusually large amount of phototherapy. No evidence for an associated porphyria has been obtained. We believe this is a unique case because transient porphyrinemia associated with neonatal blistering does not appear to have been reported previously. Furthermore, blistering associated with phototherapy is rare.
To test the noninvasive monocyte monolayer assay in predicting hemolytic severity in utero, we studied 18 patients from two institutions with significant erythrocyte alloantibodies. Serum samples were obtained from each patient. Each subject donated a serum sample during her pregnancy. Monocytes were harvested from a single healthy donor and grown in monolayer culture. Erythrocytes with the appropriate antigens were sensitized with maternal serum and incubated with the monocyte monolayers. Erythrophagocytosis was scored as a percentage of the positive control. Perinatal outcomes were assessed post hoc and cases were classified as unaffected or mildly, moderately, or severely affected using standard definitions. Prenatal management was conducted without knowledge of the results. Six of the 18 patients had severe disease and one of the 18 had moderately severe disease. At a cutoff level of 20, the assay generated the following results: sensitivity was 7 of 7 (100%), specificity was 10 of 11 (90.9%), positive predictive value was 7 of 8 (87.5%), and negative predictive value was 10 of 10 (100%). The monocyte monolayer assay appears to be a useful, noninvasive modality for predicting the severity of hemolytic disease in utero.