Objective To investigate the application of multiple ligation-dependent probe amplification (MLPA) in allele detection of 2 rare Rh partial D individuals.Methods 2 patients' peripheral blood samples were collected from local hospital in April and September 2012 for RhD phenotyping.Genomic DNA was extracted from peripheral blood after the Rhesus D phenotype had been identified.Multiple ligation dependent probe amplification (MLPA) was used to determinate the RHD and RHCE gene,then the data including gene copy numbers,point mutation or deletion and hybrid fusion were analyzed by GeneMarker software.Results coming from polymerase chain reaction-sequence specific primer (PCR-SSP) and conventional serologic methods were compared with the results of MLPA.Results The MLPA results showed the copy numbers of RHD and RHCE gene of the subject samples as following:RHD was 1.0,RHC and RHc were 1.0,RHE was 0,RHe was 3.0.The copy numbers of D03-380T,D03-455A,D04 602C,D05-667T,D04-514A,D05-787G were 0.Homozygous deletion were found in exon 3 5 of RHD gene.Deletion of exons 3-5 of RHD gene were found in PCR-SSP while alleles of C,c and e gene were found to be positive.Serologic results indicated that RBC of subject samples could be agglutinated with monoclonal anti-D in tubes and the agglutination strength was 2+,serologic typing results of Rh antigens were C+c+E-e+.The PCR-SSP results and serologic results were basically consistent with MLPA.Conclusions The RhD phenotype of the 2 individuals were D Ⅵ type 4,which was found in Chinese for the first time.As MLPA could detect known point mutation,deletion,hybridization and copy numbers of gene,so it can be used to determine the allele of variant RhD.

Background: The RHCE allele is highly polymorphic and many variants have been described, especially in individuals of African origin. Donors carrying these variants can be falsely typed and elicit transfusion reactions, and patients carrying such a variant may be at risk to develop allo-antibodies in response to mismatched transfusions. Not much is known about the frequency of RHCE variants in Chinese populations, whereas in China genotyping assays are increasingly applied for typing of blood donors and patients.

BackgroundThe para‐Bombay phenotype is characterized by the absence or weak expression of ABH antigens on the surface of red blood cells, but normal expression in saliva.Study Design and MethodsThe para‐Bombay phenotype of the nine Chinese probands was identified by standard serologic techniques. The coding regions of FUT1 and FUT2 genes were amplified by polymerase chain reaction and then directly sequenced. ABO genotyping was performed by polymerase chain reaction with sequence‐specific priming method. The FUT1 and FUT2 genotypes and the distribution in all reported Chinese para‐Bombay individuals including our study were also summarized.ResultsFive FUT1 genotypes, h1h3 (n = 3), h1h2 (n = 3), h1h1 (n = 1), h3h3 (n = 1), and h2h3 (n = 1), and three functional FUT2 genotypes, Se357Se357 (n = 4), Se357Se357, 716 (n = 4), and Se357Se357, 385 (n = 1) described before were identified in nine probands.ConclusionsThe review of the literature shows that a total of 17 FUT1 alleles and four FUT2 alleles (Se357, Se357,716, Se357 385, Se) have been identified in Chinese para‐Bombay individuals. The four FUT1 alleles, h1 (547delAG), h2 (880delTT), h3 (C658T), and h4 (C35T; A980C) are most prevalent, which account for more than 90% of all allele counts and are essential to be involved when developing para‐Bombay genotyping kit for Chinese.