Schousboe Peter, Verder Henrik, Jessen Torben E, Heiring Christian, Bender Lars, Ebbesen Finn, Dahl Marianne, Eschen Christian, Fenger-Grøn Jesper, Höskuldsson Agnar, Reinholdt Jes, Scoutaris Nikolaos, Smedegaard Heidi
Department of Paediatrics, Holbaek University Hospital, Holbaek, Denmark.
Department of Clinical Biochemistry, Holbaek University Hospital, Holbaek, Denmark.
Acta Paediatr. 2020 Feb;109(2):280-284. doi: 10.1111/apa.14896. Epub 2019 Jul 14.
To develop a fast bedside lung maturity test.
Gastric aspirates obtained from premature infants contain lamellar bodies, carrying lung surfactant. To estimate lung maturity, we isolated lamellar bodies from fresh gastric aspirates by centrifugation. Erythrocytes and other cells were lysed by adding water and discarded subsequently with the supernatant. Mid-infrared spectroscopy was then performed to measure the lung maturity as lecithin-sphingomyelin ratio. Lecithin was determined as dipalmitoylphosphatidylcholine, the most surface-active phospholipid. Algorithms to measure lecithin and sphingomyelin concentrations in fresh gastric aspirates were developed on aspirates from 140 premature infants. Each gastric aspirate sample was divided into two samples: one for mass spectrometry as reference and one for spectroscopy. Development of the algorithm is described in detail in Appendix S1.
Gastric aspirates stored at 4-5°C avoid flocculation of proteins and phospholipids in contrast to when the aspirates were frozen and thawed. Omission of freezing and concentration of the lung surfactant by centrifugation combined with diminished influence of proteins improves the spectroscopic measurement of lecithin-sphingomyelin ratio. Measurement of lecithin-sphingomyelin ratio by the new method was performed within 10-15 minutes.
We present a new fast bedside lung maturity test on fresh gastric aspirate for early targeted surfactant treatment.
开发一种快速的床边肺成熟度检测方法。
早产婴儿的胃吸出物中含有携带肺表面活性物质的板层小体。为了评估肺成熟度，我们通过离心从新鲜胃吸出物中分离出板层小体。通过加水使红细胞和其他细胞裂解，随后与上清液一起丢弃。然后进行中红外光谱法以测量作为卵磷脂 - 鞘磷脂比率的肺成熟度。卵磷脂被确定为二棕榈酰磷脂酰胆碱，即最具表面活性的磷脂。基于140名早产婴儿的吸出物开发了测量新鲜胃吸出物中卵磷脂和鞘磷脂浓度的算法。每个胃吸出物样本被分成两个样本：一个用于质谱分析作为参考，另一个用于光谱分析。算法的开发在附录S1中有详细描述。
与冷冻和解冻吸出物相比，在4 - 5°C下储存的胃吸出物可避免蛋白质和磷脂的絮凝。通过离心省略冷冻和浓缩肺表面活性物质，同时减少蛋白质的影响，可改善卵磷脂 - 鞘磷脂比率的光谱测量。用新方法测量卵磷脂 - 鞘磷脂比率在10 - 15分钟内完成。
我们提出了一种基于新鲜胃吸出物的新型快速床边肺成熟度检测方法，用于早期靶向性表面活性物质治疗。
