Publication details
Two Reliable Methodical Approaches for Non-Invasive RHD Genotyping of a Fetus from Maternal Plasma
| Authors | |
|---|---|
| Year of publication | 2020 |
| Type | Article in Periodical |
| Magazine / Source | Diagnostics |
| MU Faculty or unit | |
| Citation | |
| Web | https://www.mdpi.com/2075-4418/10/8/564 |
| Doi | http://dx.doi.org/10.3390/diagnostics10080564 |
| Keywords | non-invasive fetal genotyping; RHDgene; cell-free fetal DNA; real-time PCR; QF PCR; Rh blood group system; red blood cell alloimmunization; hemolytic disease of the fetus and newborn |
| Description | Noninvasive fetalRHDgenotyping is an important tool for predicting RhD incompatibility between a pregnant woman and a fetus. This study aimed to assess a methodological approach other than the commonly used one for noninvasive fetalRHDgenotyping on a representative set of RhD-negative pregnant women. The methodology must be accurate, reliable, and broadly available for implementation into routine clinical practice. A total of 337 RhD-negative pregnant women from the Czech Republic region were tested in this study. The fetalRHDgenotype was assessed using two methods: real-time PCR and endpoint quantitative fluorescent (QF) PCR. We used exon-7-specific primers from theRHDgene, along with internal controls. Plasma samples were analyzed and measured in four/two parallel reactions to determine the accuracy of theRHDgenotyping. TheRHDgenotype was verified using DNA analysis from a newborn buccal swab. Both methods showed an excellent ability to predict theRHDgenotype. Real-time PCR achieved its greatest accuracy of 98.6% (97.1% sensitivity and 100% specificity (95% CI)) if all four PCRs were positive/negative. The QF PCR method also achieved its greatest accuracy of 99.4% (100% sensitivity and 98.6% specificity (95% CI)) if all the measurements were positive/negative. Both real-time PCR and QF PCR were reliable methods for precisely assessing the fetalRHDallele from the plasma of RhD-negative pregnant women. |