Amniotic Fluid

By: Cabadin, Cayetano, Chumacera,

and Detras
Amniotic Fluid

❏ Product of fetal metabolism

❏ Fetal distress and fetal maturity
❏ Present in the amnion
❏ Membranous sac
❏ Metabolically active
❏ Produce peptides, growth factors, and cytokines
Primary Functions

❏ Provide protective cushion for the fetus

❏ Allow fetal movement
❏ Stabilize temperature
❏ Permit proper lung development
Volume

❏ is regulated by a balance between

❏ production of fetal urine and lung fluid
❏ absorption from fetal swallowing and intramembranous flow
❏ 800 to 1200 mL
❏ Polyhydramnios
❏ Oligohydramnios
Volume

❏ First trimester: maternal circulation

❏ Latter third to half: lung liquid
❏ Lung surfactants
❏ Fetal urine and fetal swallowing
❏ Polyhydramnios
❏ Oligohydramnios
Chemical Composition

❏ Placenta
❏ Similar to maternal plasma
❏ Small amount of sloughed fetal cells
❏ Biochemical substances
❏ Alpha-fetoprotein
❏ acetylcholinesterase
Chemical Composition

❏ Fetal urine production

❏ inc: creatinine, urea, UA
❏ dec: glucose, proteins
❏ Amniotic fluid creatinine
❏ 1.5-2.0 mg/dL
❏ above 2.0 mg/dL
Differentiating Maternal Urine from
Amniotic Fluid

❏ Determine possible premature membrane rupture or accidental

puncture or maternal bladder during specimen collection
❏ Chemical analysis of
❏ Creatinine and urea
❏ Glucose and protein
❏ Fern test
Specimen Collection

❏ Indications for Amniocentesis

❏ Recommended for neural tube defects
❏ To detect genetic disorders or evaluate the
health of the fetus
❏ Fetal body measurements (aided by ultrasonography)
❏ Fetal epithelial cells
❏ Indicate genetic material and biochemical
substances
Amniocentesis

❏ Safe procedure
❏ Performed after 14th week of gestation
❏ Fluid for chromosome analysis (16 weeks of
gestation)
❏ Tests for intrauterine growth retardation (end of 2nd
trimester)
❏ Tests for fetal distress and maturity (3rd trimester)
❏ Transabdominal amniocentesis (most common)
Collection

❏ Vaginal amniocentesis (greater risk of infection)

❏ Maximum of 30 ml
❏ First 2 or 3 ml (can be contaminated)
❏ Fluid for bilirubin analysis (diagnosis of HDN)
❏ Protected from light at all times
Specimen Handling and Processing

❏ Vary with the test requested and with the

methodology used by the lab
❏ Fluid for fetal lung maturity
❏ Bilirubin testing
❏ Cytogenetic/Microbial studies
❏ Fluid for chemical testing
Color and Appearance

❏ Normal amniotic fluid = colorless, slight to

moderate turbidity
❏ Blood streaked - traumatic tap
❏ Kleihauer-Betke Test
❏ Yellow colored - presence of bilirubin (HDN)
❏ Very Dark red-brown fluid - fetal death
❏ Meconium - first bowel movement
❏ May result due to fetal distress
❏ Dark green, mucus-like material
Tests for Fetal Distress

HDN (Hemolytic Disease of the Newborn)

❏ Evaluates the severity of fetal anemia produced by

HDN
❏ Incidence is decreasing rapidly
❏ Amniotic fluid bilirubin - measured by
spectrophotometer
❏ Optical density - measured between 365 - 550 nm
❏ Absorbance difference at 450 nm
Cont. of HDN

❏ Liley graph plots the absorbance difference at

450 nm
❏ Zone 1 - mildly affected fetus
❏ Zone 2 - moderate hemolysis and
require careful monitoring anticipating
an early delivery
❏ Zone 3 - severe hemolysis and severely
affected fetus
Cont. HDN

❏ Specimens contaminated with meconium

❏ Falsely low A450 values and not acceptable
❏ Specimens contaminated with blood
❏ Not acceptable because of the maximum absorbance of oxyhemoglobin
❏ Interference can be removed by extraction with chloroform
Neural Tube Defects

❏ Detected by the presence of

❏ Maternal serum alpha-fetoprotein (AFP)
❏ Ultrasound
❏ Amniocentesis
❏ Indicated by increased levels of AFP
❏ Examples
❏ Spina bifida
❏ Anencephaly
Neural Tube Defects

❏ AFP
❏ Produced by the fetal liver
❏ Increased levels in NTD
❏ Normal values are based on the week of gestational age
❏ Reported in multiples of median (MoM)
❏ A value 2x greater than the median value is abnormal
❏ High levels of AFP → measure amniotic acetylcholinesterase
Fetal Lung Maturity

❏ Respiratory distress syndrome

❏ Caused by an insufficiency of lung surfactant production
or an anatomical defect
❏ Atelectasis
❏ Hypoxemia
❏ Surfactant
❏ Prevents the collapse of the alveoli
❏ Decrease surface tension
❏ Amount of surfactant in fetal lungs
❏ Measured by amount of surfactant in amniotic fluid
Lecithin-Sphingomyelin Ratio

Lecithin
❏ Primary component of surfactant
❏ Produced at a relatively low and constant rate until 35th week of gestation
Sphingomyelin
❏ Produced at a constant rate at about 26 weeks’ gestation
L:S ratio -> <1.6 prior to 35th week of gestation
-> >2.0 to prevent alveolar collapse
❏ Falsely elevated: with blood or meconium
Phosphatidyl glycerol

❏ Lung surface lipid

❏ Essential for adequate lung maturity
❏ Detected after 35th week gestation

Aminostat-FLM

❏ Polyclonal anti-PG bodies specific for PG-containing lamellar bodies

❏ Negative= Pulmonary immaturity
❏ Positive=Pulmonary maturity
 Foam stability index
❏ Foam or Shake test
❏ For individual lung-surface lipid concentration
❏ Amniotic fluid+95% etOH

(+)continuous line of bubbles

❏ Semiquantitative measurement

Gradient of ethanol/fluid (0.42 ml to 0.55ml in 0.01ml increment)

value= >47 means (+) for FLM

Lamellar bodies
❏ 90% phospholipid
❏ 10% protein
❏ Type II pneumocytes of the fetal
lung @ 24th week
❏ 26th week
❏ No. of lamellar bodies=amt of phospholipid in the fetal lungs
Lamellar body count
❏ Platelet channels of automated hematology counters
❏ Impedance or optical methods

Advantage of LBC:
1. Rapid turnaround time
2. Low reagent cost
3. Wide availability
4. Low degree of technical difficulty
5. Low volume of amniotic fluid required
6. Excellent clinical performance