Physiology • Amnion: a membranous sac that surrounds the fetus • Amniotic fluid – Provides protective cushion for fetus – Allows fetal movement – Stabilizes fetal temperature exposure – Permits proper lung development – Exchanges water and chemicals among the fluid, fetus, and maternal circulation
Chemical Composition • Composition similar to maternal plasma with sloughed fetal cells – Cells are used for cytogenetic analysis • The fluid also contains biochemical substances that are produced by the fetus, such as bilirubin, lipids, enzymes, electrolytes, urea, creatinine, uric acid, proteins, and hormones • Fetal urine increases creatinine, urea, and uric acid • Fetal age can be estimated by creatinine – <36 weeks = 1.5 to 2.0 mg/dL – >36 weeks = >2.0
• Needed to determine premature membrane rupture or
accidental puncture of maternal bladder from amniocentesis • Measure creatinine, glucose, protein, and urea – Amniotic fluid has <3.5 mg/dL creatinine and <30 mg/dL urea – Values as high as 10 mg/dL for creatinine and 300 mg/dL for urea may be found in urine – The presence of glucose, protein, or both is associated more closely with amniotic fluid • Fern test: specimen air dries on glass slide; examine microscopically for “fern-like” amniotic fluid crystals
• Collection of fetal epithelial cells in amniotic fluid
indicate the genetic material of the fetus – Examined for chromosome abnormalities by • Karyotyping • Fluorescence in situ hybridization (FISH) • Fluorescent mapping spectral karyotyping • DNA testing • Biochemical substances that the fetus has produced – Analyzed by thin-layer chromatography
Tests for Fetal Distress • HDN – Most commonly Rh-negative mothers – Other red blood cell (RBC) antigens can also produce HDN – Fetal cells with antigens enter maternal circulation and cause production of maternal antibodies – Maternal antibodies cross the placenta and destroy fetal cells with the corresponding antigen
– Alpha-fetoprotein (AFP) produced by the fetal liver prior to 18 weeks’ gestation – Increased levels in maternal blood or amniotic fluid indicate possible anencephaly or spinal bifida – Increased levels are found when skin fails to close over neural tissue – Measure maternal blood first, then amniotic fluid
(maximum AFP 12 to 15 weeks) • Report multiples of the median (MoM) – Median is laboratory’s reference level for a given week of gestation – More than two times the median (MOM) is abnormal • Follow with fluid amniotic acetylcholinesterase (AChE): more specific for neural disorders – Do not perform on a bloody specimen
respiratory distress syndrome (RDS) • Lack of lung surfactant, which keeps the alveoli open during inhaling and exhaling • Surfactant decreases the surface tension on the alveoli so they can inflate more easily • Many laboratory tests are available for FLM
Lecithin-Sphingomyelin (L/S) Ratio • Considered the reference method • Lecithin is the primary component of the lung surfactants; increased production occurs after the 35th week • Sphingomyelin is produced at a constant rate after the 26th week and serves as a control for the rise in lecithin • L/S ratio is 1.6 prior to week 35 and rises to 2.0 or greater for alveolar stability after week 35 • Therefore, preterm delivery is considered safe with an L/S ratio of 2.0 or higher • Test is performed using thin-layer chromatography • Many laboratories have replaced the L/S ratio with the quantitative phosphatidyl glycerol immunoassays and lamellar body density procedures
• Lamellar body diameter ranges in size from 1.7 to 7.3 fL or 1 to 5 µm
• LBCs can be obtained using the platelet channel of automated hematology analyzers • Advantages of LBC – Rapid turnaround time – Low reagent cost – Wide availability – Low degree of technical difficulty – Low volume of amniotic fluid required – Excellent clinical performance • Specimens contaminated with meconium or mucous cannot be used
Protocol for Performing LBC 1. Mix the amniotic fluid sample by inverting the capped sample container five times. 2. Transfer the fluid to a clear test tube to allow for visual inspection. 3. Visually inspect the specimen. Fluids containing obvious mucus, whole blood, or meconium should not be processed for an LBC. 4. Cap the tube and mix the sample by gentle inversion or by placing the test tube on a tube rocker for 2 minutes. 5. Flush the platelet channel; analyze the instrument’s diluents buffer until a background count deemed acceptable by the laboratory is obtained in two consecutive analyses. 6. Process the specimen through the cell counter and record the platelet channel as the LBC.