Genetic diagnosis of MN and ABO hemolytic disease of the newborn complicated with pyruvate kinase deficiency: a case report and literature review / 中国输血杂志

【Objective】 To investigate the clinical and genetic characteristics of hemolytic disease of the newborn(HDN) induced by anti-M complicated with pyruvate kinase deficiency (PKD) disease. 【Methods】 The clinical data of a pregnant woman with unexplained adverse pregnancy outcome in the third trimester were retrospectively analyzed, and neonate anemia status and blood transfusion were followed up. With informed consent, peripheral blood of the neonate and her parents were collected for serological tests and disease-related gene target sequence capture and sequencing. The clinical and genetic characteristics of HDN induced by anti-M or PKD were reviewed. 【Results】 The neonate presented severe anemia, hepatosplenomegalism, hyperbilirubinemia at birth, and was confirmed MN combined with ABO neonatal hemolysis by serological tests. The neonate recovered by receiving blood exchange and phototherapy treatments, but he needed to receive blood transfusion each month because of hemolytic anemia. Genetic analysis showed that the neonate had compound heterozygous mutations (c. 1096C>T; c. 941T>C) of PKLR gene inherited from her parents. 【Conclusion】 The clinical manifestations of PKD are similar to that of HDN caused by anti-M in the early stage of the disease. Clinicians should exclude the metabolic diseases of red blood cells when diagnosing severe HDN caused by anti-M.

Novel mutations associated with pyruvate kinase deficiency in Brazil

Abstract Background: Pyruvate kinase deficiency is a hereditary disease that affects the glycolytic pathway of the red blood cell, causing nonspherocytic hemolytic anemia. The disease is transmitted as an autosomal recessive trait and shows a marked variability in clinical expression. This study reports on the molecular characterization of ten Brazilian pyruvate kinase-deficient patients and the genotype-phenotype correlations. Method: Sanger sequencing and in silico analysis were carried out to identify and characterize the genetic mutations. A non-affected group of Brazilian individuals were also screened for the most commonly reported variants (c.1456C>T and c.1529G>A). Results: Ten different variants were identified in the PKLR gene, of which three are reported here for the first time: p.Leu61Gln, p.Ala137Val and p.Ala428Thr. All the three missense variants involve conserved amino acids, providing a rationale for the observed enzyme deficiency. The allelic frequency of c.1456C>T was 0.1% and the 1529G>A variant was not found. Conclusion: This is the first comprehensive report on molecular characterization of pyruvate kinase deficiency from South America. The results allowed us to correlate the severity of the clinical phenotype with the identified variants.

Genetic diagnosis of erythrocyte pyruvate kinase deficiency and discovery of PKLR gene novel mutations / 中华实用儿科临床杂志

Objective To investigate the clinical symptoms of pyruvate kinase deficiency (PKD) and the new mutation type of PKLR gene in 3 cases of PKD,and to explore the method for PKD gene diagnosis.Methods Sequencing of blood system-related genes in 3 children was performed by target sequence capture and high-throughput sequencing technology,and the protein function of mutant gene was forecasted,after detecting the pathogenicity of the patients,these genotypes were confirmed by Sanger sequencing.Results In the 3 children,5 types of PKLR gene mutations were found:double heterozygous mutations c.1529G ＞ A(p.R510Q) and c.1031T ＞ G(p.I344S),homozygous mutation c.847G ＞ T (p.V283F),double heterozygous mutations c.979delC (p.L327fs)and c.604_617del (p.V202fs).PKLR gene c.1529G ＞ A(p.R510Q) mutation had been reported previously,and the other four mutations were new.c.1031T ＞ G (p.I344S) and c.847G ＞ T (p.V283F) was possibly pathogenic mutation,which meant that the probability of mutation of this gene was more than 90％ and c.979delC (p.L327fs) and c.604 _617del (p.V202fs) variation was a pathogenic variation.These 5 mutations had a greater effect on protein function,and all ofthem were pathogenic mutations.Conclusion Since PKD patients are difficult to be diagnosed clinically,PKLR gene variation can be detected by target sequence capture and high throughput sequencing technology,and the pathogenicity of the new mutant is evaluated.