Prenatal RHD gene determination and dosage analysis by PCR: clinical evaluation.

BACKGROUND: Use of the polymerase chain reaction (PCR) for detection of the RHD gene can measure the RHD gene status for unborn babies at risk for hemolytic disease of the newborn (HDN). The occurrence of D gene variants has led to errors in prenatal typing. Previous reports have highlighted the danger of assigning a positive fetus as negative, resulting in intrauterine fetal deaths. OBJECTIVE: To evaluate the effectiveness of a testing strategy whereby PCR was not only performed to determine the presence/absence of the RHD gene, but also used to assess the D gene copy number (zero, one or two RHD genes) in family studies for at risk pregnancies. METHODS: Samples comprising maternal (57) and paternal (42) peripheral blood samples, amniotic fluid (64), and matching cord blood (64) were collected. Rhesus (Rh) serotyping was performed on all blood samples. For RHD genotyping, DNA was extracted from all samples except for 28 cord samples, where only serotyping was performed (total 199 DNA genotyping). RHD gene PCR amplified exon 4 and exon 7 regions of the RHD gene. The dosage of RHD gene was determined by comparing the intensity of the RHD gene to that of the RHCE gene. RESULTS: A total of 197/199 samples showed concordance between exon 4 and exon 7 PCR results. Two discrepant results occurred in one family: the father carried one normal D gene and one D gene variant where PCR was tested to be positive using exon 4 but negative using exon 7. One of a pair of dizygotic twins inherited this abnormal D gene and was mildly affected by HDN. This was correctly identified antenatally and the pregnancy successfully managed. The concordance rate between serotypes and genotypes for 135 blood samples was 100%. Amongst the family groups, 8/14 heterozygous fathers transmitted the D gene and 26/26 homozygous fathers transmitted the D gene to the babies. The concordance rate between RHD genotypes from amniotic fluid and Rh D serotypes from cord blood was also 100%. CONCLUSION: The present study demonstrates the effectiveness of using PCR in a clinical setting. It verifies the importance of testing more than one region of the gene, and also the need for a testing strategy where both maternal and paternal testing for RHD gene dosages are performed.

Vacuum extraction: a randomized controlled comparison of the New Generation cup with the original Bird cup.

A new design of vacuum extractor cup--'the New Generation cup'--has recently been introduced into clinical practice. Its major modification is a traction cord which passes around the rim of the cup for 180 degrees and is free to slide within the rim. Claims that this design 'enables the operator to pull obliquely without causing the cup to tilt' and thereby 'reduces failure rates, reduces the incidence of scalp trauma and increases operator confidence' have been evaluated in a randomized controlled trial. 123 women with singleton pregnancies of 37 completed weeks or more, with a cephalic presentation and for whom a decision to deliver by vacuum extraction had been taken, were randomly allocated to the 'New Generation' cup or BIRD's original vacuum extractor cup; 50 mm anterior and posterior cups were used in both groups as appropriate. The two groups were comparable at entry and delivered by obstetricians of similar status. The two types of cup were similar in respect of number of failures to deliver with the vacuum extractor, correct positioning of the cup, number of pulls required for delivery and time taken to expedite delivery. Cup detachments occurred in nine cases allocated to the 'New Generation' cup compared with four allocated to the original BIRD cup. The babies in the two groups were in similar condition at birth and sustained similar amounts of scalp trauma. Neonatal jaundice, both clinical and biochemical, was more common in babies delivered with the 'New Generation cup' and this was reflected in greater use of phototherapy in this group. Operators were equally divided in their preference of cup.(ABSTRACT TRUNCATED AT 250 WORDS)

Follow up of babies delivered in a randomized controlled comparison of vacuum extraction and forceps delivery.

Babies delivered in a randomized controlled comparison of vacuum extraction vs. forceps delivery were reassessed at 9 months of age. There were no statistically significant differences between the two groups as regards head circumference, weight, or head circumference to weight ratio, nor in the results of hearing and vision tests. The reasons for hospital readmission, pediatric follow-up and parents' and health visitors' worries appeared to be unrelated to the mode of delivery in nearly all cases. The finding in the original trial that neonatal jaundice was more common following vacuum extraction was reinforced by an additional case of jaundice in the vacuum extractor group which had necessitated readmission to hospital.