Clinical utility of next-generation sequencing in the diagnosis of hereditary haemolytic anaemias.

Hereditary haemolytic anaemias are genetically and phenotypically heterogeneous disorders characterized by increased red cell destruction, with consequences ranging from innocuous to severe life-threatening anaemia. Diagnostic laboratories endeavour to assist clinicians reach the exact patient diagnosis by using tests principally based on morphological and biochemical techniques. However, these routine studies may be inconclusive, particularly in newborn infants and when transfusions have recently been administered. Large numbers and size of the potentially involved genes also impose a practical challenge for molecular diagnosis using routine sequencing approaches. To overcome these diagnostic shortcomings, we have utilized next-generation sequencing to provide a high-throughput, highly sensitive assay. We developed a panel interrogating 28 genes encoding cytoskeletal proteins and enzymes with sequencing coverage of the coding regions, splice site junctions, deep intronic and regulatory regions. We then evaluated 19 samples, including infants with unexplained extreme hyperbilirubinaemia and patients with transfusion-dependent haemolytic anaemia. Where possible, inheritance patterns of pathogenic mutations were determined by sequencing of immediate relatives. We conclude that this next-generation sequencing panel could be a cost-effective approach to molecular diagnosis of hereditary haemolytic anaemia, especially when the family history is uninformative or when routine laboratory testing fails to identify the causative haemolytic process.

Can we cut the incidence of necrotizing enterocolitis in half--today?

Necrotizing enterocolitis (NEC) is a common gastrointestinal emergency of neonates. Population studies estimate the incidence of NEC at between 0.3 and 2.4 per 1000 live births in the United States, with a predominance of cases among preterm neonates born at the earliest gestational ages. The disease burden of NEC includes an overall disease-specific mortality rate of 15-20%, with yet higher rates in those of earliest gestations. The NEC burden also includes an increase in hospital costs approximating $100,000/case, as well as severe late sequellae including parenteral nutrition-associated liver disease and short bowel syndrome. Differentiating NEC from other forms of acquired neonatal intestinal disease is critical to assessing the success of NEC prevention strategies. Promising new prevention strategies are now being tested; one such is prophylactic heparin-binding epidermal growth factor-like growth factor (HB-EGF) administration. However, two prevention strategies have already been shown in meta-analyses to reduce the incidence of NEC, but we speculate that these are not being fully utilized. They are; 1) implementing a written set of feeding guidelines (also called standardized feeding regimens) for newborn intensive care unit (NICU) patients, and 2) implementing programs to increase the availability of human milk for patients at risk of developing NEC.