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A 26-year-old white woman had premature rupture of membranes at 35 weeks' gestation. Cervical specimens initially demonstrated group B streptococci and Chlamydia trachomatis. Amniocentesis was performed and the amniotic fluid was positive for chlamydia by direct fluorescent antibody stain. The Gram stain was negative. The patient had an elevated white blood cell count. Labor was induced because of suspected chorioamnionitis. A 2120-g female infant was delivered with an Apgar score of 9 at 1 and 5 minutes. The infant's eye and nasopharyngeal specimens were positive for chlamydia by direct fluorescent antibody stain. After delivery, both Ureaplasma urealyticum and C trachomatis were isolated from the amniotic fluid and fetal membranes. This is the first reported case of chlamydial isolation in amniotic fluid.

A test that circumvents the complex methodology needed for the determination of L/S ratios has been developed. It has been observed that the optical density, measured at 400 nm, of supernatants collected from fresh amniotic fluids centrifuged at 2000 g for 10 minutes correlates with L/S ratios.

Amniotic fluids obtained by amniocentesis at 16 weeks to term were examined for the presence of Chlamydia trachomatis, Mycoplasma hominis, and Ureaplasma urealyticum. Of 140 fluids tested, none harbored chlamydiae, and only one harbored mycoplasma, M. hominis. A number of amniotic fluids were subsequently tested for their ability to inhibit the growth of these microorganisms. Amniotic fluids and chlamydial suspensions in a 2:1 ratio were incubated 30 to 90 minutes before their inoculation in McCoy cells. Procedures were followed for chlamydial isolation. Genital mycoplasmas were incubated with amniotic fluid samples for 24 or 48 hours at 35 degrees C. Growth in amniotic fluid specimens was compared with growth in pseudoamniotic fluid and broth controls. Fourteen amniotic fluid specimens collected from gestations of 16 to 40 weeks, were found to be inhibitory to the formation of inclusions of C. trachomatis in McCoy cells. Ten amniotic fluid specimens (16 to 39 weeks, gestation) demonstrated various degrees of inhibition against M. hominis, and three fluids were inhibitory to the growth of Ureaplasma. The inhibitor was heat and protease resistant and activity was proportional to concentration. The molecular weight of the inhibitor was probably greater than 10,000 daltons, and pH, although perhaps a contributing factor, was not the cause of the inhibition.

In this study, we have attempted to correlate optical density measurements of amniotic fluids with L/S ratios. We may conclude, with over a 98 per cent accuracy, that fluids having optical density readings of 0.15 and above, at 650 nm. will have an L/S ratio over 2.0. Fluids having optical density readings up to 0.05 will have L/S ratios of about 1.3. Finally, amniotic fluids having optical densities greater than 0.05 and less than 0.15 will have L/S ratios of approximately 1.5.

Optical density readings of amniotic fluids of 0.15 or greater at 650 nm have been noted to correlate with fetal pulmonary maturity. The amniotic fluid absorbance has been shown to be due not only to lecithin and sphingomyelin but also to other surfactants, including phosphatidyl glycerol and inositol. The addition of lecithin and sphingomyelin to previously centrifuged amniotic fluid (i.e., optical) density less than 0.001: L/S ratio, nondetectable) results in an increase in absorbance. At any simulated L/S ratio, the addition of phosphatidyl glycerol and/or phosphatidyl inositol results in a further increase in optical density. It is suggested that optical density readings represent more closely the surfactant composition of amniotic fluid than L/S ratios; therefore, it appears that optical density measurements are a better predictor of fetal pulmonary development than are L/S ratios.

A simple, rapid, economical, and accurate test to assess fetal pulmonary maturity on a 24-hour basis, 7 days a week, is urgently needed. A 15-minute test has been developed that correlates well with fetal pulmonary maturity. Optical density (OD) readings at 650 nm of greater than or equal to 0.15 in pigment-free, centrifuged (2000 Xg, 10 minutes) amniotic fluids obtained from gestations of 33 to 42 weeks correlate with fetal pulmonary maturity (131 of 131, 100%). When OD readings are less than 0.15 in fluids from gestations of 24 to 40 weeks, and antepartum steroids are utilized, the incidence of respiratory distress syndrome (RDS) is 8.3% (14 of 169). The true false-negative rate in this group is therefore unknown.

A simple and rapid test that correlates with L/S ratios has been developed. By centrifuging fresh, unfrozen amniotic fluids at 200g for 10 minutes and measuring the optical density at at 650 nm, correlation with L/S ratios is obtained. Optical density readings of 0.15 or greater correlate 100% with L/S ratios of 2.0 or greater. Optical readings below 0.15 correlate 94% with L/S ratios below 2.0.

Fetal pulmonary maturity is generally determined by analyzing amniotic fluid for surfactants. This task is accomplished by lipid extraction of the fluid and resolution, identification and quantitation of the isolated lipids with thin-layer chromatography. These methods are lengthy, cumbersome and often not available on demand. A quick, simple, reliable and economical test therefore would be highly desirable. We have been able to correlate an optical density (OD) reading of amniotic fluid at 650 nm greater than or equal to 0.15 with the absence of hyaline membrane disease (HMD). For 428 fluids in which an OD reading of greater than or equal to 0.15 was found and delivery occurred within 48 hours, HMD was present in only two infants. The accuracy of the test was 99.53% , with a false-positive rate of 0.47%. With the use of this simple and accurate test one can satisfy the requirement of an on-demand test to determine fetal pulmonary maturity.

In this study we have attempted to correlate amniotic fluid optical density measurements and lecithin/sphingomyelin (L/S) ratios with the presence of phosphatidylglycerol. An 85% correlation (126 of 148) of all three parameters was noted. When the assumption was made that phosphatidylglycerol is indicative of fetal pulmonary maturity, 93 of 94 fluid samples having an optical density at 650 nm greater than or equal to 0.15 had demonstrable phosphatidylglycerol for a 1% false positive rate. In 53 fluid samples having an optical density at 650 nm less than 0.15, phosphatidylglycerol was present in 11 and absent in 42, a 21% false negative rate. The same analyses with lecithin/sphingomyelin ratios showed a 5% false positive rate and a 31% false negative rate.