ABSTRACT
The Rh blood grouping system is a critical standardized test in transfusion medicine,espe-cially for the cases related to haemolytic transfusion reactions and neonatal haemolytic disease caused by clinical RhD blood group incompatibility.In the present case report,we presented two cases with the un-common RHD gene variation RHD*DEL37.The blood samples of the two subjects were mistakenly iden-tified as RhD-negative through conventional serological testing.Firstly,both blood samples were tested negative for the RhD antigen using traditional tube test and gel microcolumn methods.The phenotyping of RhCE were identified as ccEe and ccee for each sample,respectively.Secondly,genetic analysis was performed using polymerase chain reaction-sequence specific prime(PCR-SSP)which revealed that nei-ther sample belonging to the several common RHD gene variants which was found in Asia.Moreover,they turned out to be positive for the RHD haplotype,which indicated that exons 1-10 on one of the RHD al-leles were entirely absent.In addition,a T＞C mutation was observed at bases 1154-31 in intron 8 of the other allele,which was located at the intron 8 breakpoint.This result was obtained after further Sanger sequencing of exons 1-10 of the RHD gene.The mutant allele was designated as RHD*DEL37 by the International Society of Blood Transfusion(ISBT)and was identified as D-elute(Del)by phenotype ana-lysis.Both samples were genotyped as RHD*DEL37 and showed positive results.In summary,the true genotype of the two blood samples,of which the screening results only using serological testing method was negative,were RHD*DEL37/RHD-(RHD*01N.01).Notably,this kind of genotype was reported for the first time in Chinese population.Moreover,the two individuals did not have ties of consanguinity,indicating that some of the Chinese individuals could be carriers of the genetic mutation.Therefore,it might be necessary to further confirm the frequency of this mutation in the Chinese population and the possibility of homozygosity for this mutation.This report identifies infrequent RHD gene mutation samples by coupling molecular biology and serological methods to prevent misclassification of blood groups.Com-bining serological and molecular biology test results to determine blood group is critical in protecting pa-tients during clinical transfusion procedures.
ABSTRACT
Objective:To investigate the germline mutation status in multi-pathway in Chinese female breast cancer patients and explore their correlation with clinicopathological characteristics. Aim to enrich the database of breast cancer germline gene mutations in Chinese population and provide laboratory evidence for the application of breast cancer targeted drugs.Methods:From January 2017 to July 2019, whole blood samples were collected from 148 women (age of onset concentrated in the 24~80 years old) diagnosed pathologically with breast cancer in the Department of breast surgery, Peking University People′s Hospital. Germline mutations in HR, MMR, BER, and KDR pathway related genes were detected by next-generation sequencing. The pathogenicity interpretation was performed, and pathogenic, likely pathogenic, and mutations of uncertain significance were screened. The clinicopathological characteristics including age at the onset, luminal typing, tumor size, metastasis, and family history were analyzed, and the correlation between mutations in different pathway genes and clinicopathological characteristics was analyzed by the Chi-squared test and Fisher′s exact probability test.Results:Among the 148 patients, there were 69 cases of HR mutations (including three types of mutations, including pathogenic, likely pathogenic and uncertain significance), 16 cases of MMR mutations, 6 cases of BER mutations and 8 cases of KDR mutation. ATM mutations in the HR pathway were associated with luminal typing ( P=0.054), and patients with HER2+breast cancer were more likely to carry ATM mutations. PMS2 mutations in the MMR pathway were correlated with tumor size ( P=0.060), and patients with tumor size>50 mm were more likely to carry PMS2 mutations. KDR mutations was significantly correlated with luminal typing and family history. ( P=0.021, P=0.024). Conclusion:The mutation frequency in BER, KDR, MMR and HR pathways in Chinese breast cancer patients increased successively. Germline mutations in ATM, PMS2 and KDR genes may be involved in the development of breast cancer in the Chinese population. Multi-pathway gene detection of breast cancer can provide laboratory evidence for the use of PARP inhibitors, trastuzumab and other targeted drugs.