Academic Journal
Rapid RHD Zygosity Determination Using Digital PCR.
| Τίτλος: | Rapid RHD Zygosity Determination Using Digital PCR. |
|---|---|
| Συγγραφείς: | Sillence, KA, Halawani, AJ, Tounsi, WA, Clarke, KA, Kiernan, M, Madgett, TE, Avent, ND |
| Στοιχεία εκδότη: | England Oxford University Press (OUP) |
| Έτος έκδοσης: | 2017 |
| Συλλογή: | PEARL (Plymouth Electronic Archiv & ResearchLibrary, Plymouth University) |
| Θεματικοί όροι: | Hemizygote, Homozygote, Humans, Phenotype, Polymerase Chain Reaction, Rh-Hr Blood-Group System, Time Factors |
| Περιγραφή: | BACKGROUND: Paternal zygosity testing is used for determining homo- or hemizygosity of RHD in pregnancies that are at a risk of hemolytic disease of the fetus and newborn. At present, this is achieved by using real-time PCR or the Rhesus box PCR, which can be difficult to interpret and unreliable, particularly for black African populations. METHODS: DNA samples extracted from 58 blood donors were analyzed using 2 multiplex reactions for RHD-specific targets against a reference (AGO1)(2) to determine gene dosage by digital PCR. Results were compared with serological data, and the correct genotype for 2 discordant results was determined by long-range PCR, next-generation sequencing, and conventional Sanger sequencing. RESULTS: The results showed clear and reliable determination of RHD zygosity using digital PCR and revealed that 4 samples did not match the serologically predicted genotype. Sanger sequencing and long-range PCR (LR-PCR) followed by next-generation sequencing revealed that the correct genotypes for samples 729M and 351D, which were serologically typed as R1R2 (DCe/DcE), were R2r' (DcE/dCe) for 729M and R1r" (DCe/dcE), R0r(y) (Dce/dCE), or RZr (DCE/dce) for 351D, in concordance with the digital PCR data. CONCLUSIONS: Digital PCR provides a highly accurate method to rapidly define blood group zygosity and has clinical application in the analysis of Rh phenotyped or genotyped samples. The vast majority of current blood group genotyping platforms are not designed to define zygosity, and thus, this technique may be used to define paternal RH zygosity in pregnancies that are at a risk of hemolytic disease of the fetus and newborn and can distinguish between homo- and hemizygous RHD-positive individuals. |
| Τύπος εγγράφου: | article in journal/newspaper |
| Περιγραφή αρχείου: | 1388-1397; Print-Electronic; application/pdf |
| Γλώσσα: | English |
| ISSN: | 0009-9147 1530-8561 |
| Relation: | E-ISSN:1530-8561; http://hdl.handle.net/10026.1/9580 |
| DOI: | 10.1373/clinchem.2016.268698 |
| Διαθεσιμότητα: | http://hdl.handle.net/10026.1/9580 https://doi.org/10.1373/clinchem.2016.268698 |
| Αριθμός Καταχώρησης: | edsbas.47593635 |
| Βάση Δεδομένων: | BASE |
| ISSN: | 00099147 15308561 |
|---|---|
| DOI: | 10.1373/clinchem.2016.268698 |