Comparison of infrared spectroscopic and fluorescence depolarization assays for fetal lung maturity

Infrared spectroscopic analysis of amniotic fluid was recently shown to be a potential useful method for the determination of fetal lung maturity. Those studies used thin-layer chromatography as a reference method for the calibration of the infrared-based technique. However, thin-layer chromatography is compromised by large intra-assay and interlaboratory coefficients of variation. Therefore in this study we have used a reference method that is based on fluorescence depolarization, the TDx FLM II assay, to verify the sensitivity and precision of infrared spectroscopy for assessment of fetal lung maturity status.

Samples of amniotic fluid were obtained by amniocentesis from 101 patients between the 24th and 40th weeks of pregnancy. Small volumes (35 microL) of amniotic fluid specimens were dried, and the infrared spectra were measured with a commercial infrared spectrometer. The fetal lung surfactant/albumin ratio was determined separately for each specimen with the TDx FLM II assay. The proposed infrared method was then calibrated and tested with a partial least-squares regression analysis to quantitatively correlate the infrared spectra with the surfactant/albumin ratios provided by the TDx FLM II assays.

A total of 144 training spectra were used to build the partial least-squares calibration model. The correlation coefficient for the training set was excellent (r = 0.92), with an SE between infrared-predicted and reference surfactant/albumin ratios of 17 mg/g. The model was then validated on a set of 69 test spectra and yielded an SE of 14 mg/g (r = 0.86). The final partial least-squares model included the 900- to 1500-cm(-1) and 2800- to 3200-cm(-1) spectral ranges and 6 partial least-squares factors.

Because the infrared-based fetal lung maturity measurements correlated well with assays from both of the current standard clinical techniques (thin-layer chromatography and fluorescence depolarization) and the procedure is less labor and training intensive, we concluded that infrared spectroscopy has the potential to emerge as the method of choice for prediction of fetal lung maturity from amniotic fluid analysis.