AF - SPECIMEN COLLECTION - 15-18 weeks OR 2nd trimester amniocentesis is for? -20-42 weeks OR 3rd trimester amniocentesis is for?

- Assess genetic defects (Ex: Trisomy 21/Down syndrome) - Fetal lung maturity (FLM), Fetal hemolytic disease (HDN)

BODY FLUIDS Flashcards by Patricia Baltazar

AMNIOTIC FLUID

Produced by the syncytiotrophoblast cells of the placenta
Peaks during 1st trimester of pregnancy (↑ blood, urine, amniotic fluid)
Composed of 2 subunits:
o Alpha = hCG, LH, FSH, TSH (identical subunits)
o Beta = Confers specificity for hCG

HUMAN CHORIONIC GONADOTROPIN (hCG)

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AMNIOTIC FLUID

what subunit of hCG ang gina gamit to test sa lab?

o Beta = Confers specificity for hCG

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AMNIOTIC FLUID - hCG

Principle:
Specimen:
Cut-off point:
Anti-hCG source:

NOTE:
-ELISA tests are very sensitive, giving positive reactions as early as ____ after conception
- Urine specimen for pregnancy testing should have a specific gravity of at least___?

Principle: Enzyme-immunoassay
Specimen: 1st morning urine
Cut-off point: 25 mIU/mL
Anti-hCG source: Rabbit
10 days
1.015 SG

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AF - PREGNANCY TESTS

CAUSE OF FALSE-POSITIVE?

CAUSE OF FALSE-NEGATIVE?

PREGNANCY TESTS

CAUSE OF FALSE-POSITIVE = Hematuria or proteinuria

CAUSE OF FALSE-NEGATIVE = Dilute urine (low S.G) – dapat first morning urine; increased SG

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Present in the amnion – a membranous sac that surrounds the fetus
source of this is the placenta

AMNIOTIC FLUID

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PRIMARY FUNCTIONS OF AMNIOTIC FLUID

Cushion for the fetus
Stabilizes temperature
Allows fetal movement
Proper lung development

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NV of AF?

Normal = 800-1,200 mL (3rd trimester)

—During 1st trimester, 35 mL of amniotic fluid is derived primarily from the maternal
circulation

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major contributor to the AF volume after the 1st trimester of pregnancy?

Fetal urine

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AMNIOTIC FLUID

CAUSES OF POLYHYDRAMNIOS = INCREASED amniotic fluid volume (>1200 mL) ?

Decreased fetal swallowing of urine
Neural tube defects (ex. Spina
bifida)

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AMNIOTIC FLUID

CAUSES OF OLIGOHYDRAMNIOS = DECREASED amniotic fluid volume (<800 mL)?

Increased fetal swallowing of urine
Membrane leakage
Urinary tract deformities

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AMNIOTIC FLUID

Method of collection of AF?

Amniocentesis — 2-3mL

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AF - SPECIMEN COLLECTION

15-18 weeks OR 2nd trimester amniocentesis is for?
-20-42 weeks OR 3rd trimester amniocentesis is for?

Assess genetic defects (Ex: Trisomy 21/Down
syndrome)
Fetal lung maturity (FLM), Fetal hemolytic
disease (HDN)

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AMNIOTIC FLUID

Quadruple screening tests prior to performing amniocentesis:

“AHUI”

o Alpha-fetoprotein
o Human chorionic gonadotropin (hCG)
o Unconjugated estriol (UE3)
o Inhibin A

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AF - SPECIMEN HANDLING

-Test for Fetal Lung Maturity, temp?
-Test for Cytogenetic Studies, temp?
-Test for HDN ?

Test for Fetal Lung Maturity = Placed on ice on delivery, kept refrigerated or frozen, –Filtration prevents loss of phospholipids – “Freeze LaMig”
Test for Cytogenetic Studies = Kept at room temperature or at 37oC
Test for HDN = Protected from light

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Detects ruptured amniotic membranes
Also used to diagnose early pregnancy (↑ estrogen)
Procedure: Specimen (Vaginal Fluid) → Slide (Air Dry) → (+) Fern-like crystals = AMNIOTIC FLUID (Due to presence of sodium chloride and
proteins)

FERN TEST

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AMNIOTIC FLUID COLOR

-Colorless/ pale yellow

-NORMAL

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AMNIOTIC FLUID COLOR

-yellow

HDN (Bilirubin), fetal distress

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AMNIOTIC FLUID COLOR

Dark-green

Meconium = 1st fetal bowel movement (sign of distress)

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AMNIOTIC FLUID COLOR

Dark red-brown

Fetal death

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AMNIOTIC FLUID

if ↑ UREA and ↑ CREA?

Maternal urine

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AMNIOTIC FLUID

if ↑ PROTEIN and ↑ GLUCOSE?

Amniotic fluid

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AF
o Most frequent complication of early delivery
o 7th most common cause of morbidity and mortality in the premature infant
o Caused by insufficiency of lung surfactant (phospholipids) production & fetal
lung immaturity

Respiratory distress syndrome

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TEST FOR FETAL LUNG MATURITY

-reference method?
-Ratio of ____ = mature fetal lungs

-Lecithin/ Sphingomyelin ratio

->2.0

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TEST FOR FETAL LUNG MATURITY

= for alveolar stability?

Lecithin

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TEST FOR FETAL LUNG MATURITY = serves as a control (due to constant production)

Sphingomyelin

TEST FOR FETAL LUNG MATURITY - Immunologic test for phosphatidylglycerol (PG) o Not affected by blood or meconium o Production of PG is delayed among diabetic mothers

AmniostatFLM

TEST FOR FETAL LUNG MATURITY that uses: - Amniotic fluid + 95% Ethanol → Shake for 15 secs →Stand for 15 mins

Foam stability (Foam/Shake test) ---- (+) Foam/Bubbles = MATURE FETAL LUNGS (Presence of phospholipids)

TEST FOR FETAL LUNG MATURITY - obsolete - Measured by fluorescence polarization, also test for FLM

Microviscosity

TEST FOR FETAL LUNG MATURITY -- a method that can be done using impedance and/or optical scatter methods since same size man gud sa PLT

Lamellar body count (LBC) ---Type ll pneumocytes produce alveolar surfactants stored in the form of lamellar bodies

TEST FOR FETAL LUNG MATURITY how many LBC indicates adequate FLM?

>32,000/uL lamellar body count = ADEQUATE FLM

TEST FOR FETAL LUNG MATURITY - ↑ Lamellar bodies = ↑ O.D. (Absorbance) - O.D. of >0.150 is equivalent to: L/S ratio of > 2.0 and the presence of PG- phosphatidyl glycerol

OD 650 nm

TEST FOR FETAL AGE amniotic fluid CREATININE if prior to 36 weeks' gestation?

1.5 to 2.0 mg/dL

TEST FOR FETAL AGE amniotic fluid CREATININE if 36 weeks (9 months?

>2.0 mg/dL

TEST FOR HDN The oldest routinely performed lab test on AF evaluates the severity of fetal anemia due to HDN

Optical Density (Absorbance) 450 -- Absorbance of amniotic fluid: o Normal = ↑ at 365nm, ↓ at 550nm o HDN = ↑ at 450 nm (bilirubin)

TEST FOR HDN - Optical Density (Absorbance) 450 Results are plotted on a Liley graph: WHAT ZONE? o Severely affected fetus (requires intervention)? o Non-affected or mildly affected fetus? o Moderately affected fetus (requires close monitoring)?

o Zone III o Zone I o Zone II

WHAT CONDITION? ("'split spine”) is a birth defect where there is incomplete closing of the backbone & membranes around the spinal cord.

Spina bifida

WHAT CONDITION? the absence of a major portion of the brain, skull, and scalp that occurs during embryonic development

Anencephaly

-Screening test for NTD? - if increased AFP? - if decreased AFP?

-Alpha-fetoprotein (AFP) -Neural tube defects -Down syndrome

Confirmatory test for Neural Tube Defect?

Confirmatory test = Acetylcholinesterase

it is the major protein produced by the fetal liver during early gestation (prior to 18 weeks)

AFP

-Not a sterile body fluid - Tracheobronchial secretions - 95% water and 5% solids other sterile BF?

-sputum - "SUB" SYNOVIAL URINE INSIDE BALDDER BLOODC

most important single component of sputum viscosity

sialic acid

Acceptable sputum specimen?

<10 S.E.C./LPF and >25 WBC/LPF alveolar macro/ dust cells

SPECIMEN COLLECTION FOR SPUTUM -most preferred sample (most concentrated; routine)? - for volume measurement? -for pediatric patients - for non-cooperative patients? - for debilitated or unconscious patients? - Specimen preservation methods

-1st morning - 24-hour sputum - Throat swab - Sputum induction - Tracheal aspiration - Refrigeration or 10% formalin

SPUTUM - MACROSCOPIC EXAMINATION Colorless or translucent?

Made up of mucus only

SPUTUM - MACROSCOPIC EXAMINATION Anchovy sauce or rusty brown

Old blood, pneumonia, gangrene

SPUTUM - MACROSCOPIC EXAMINATION Rusty (with pus)

Lobar pneumonia (S. pneumoniae)

SPUTUM - MACROSCOPIC EXAMINATION Rusty (without pus)

Congestive heart failure

SPUTUM - MACROSCOPIC EXAMINATION Currant, jelly-like

Klebsiella pneumoniae infection

SPUTUM - MACROSCOPIC STRUCTURES -Yellow or gray material, size of a pinhead; Produces foul odor when crushed -CLIN SIG: Bronchitis, bronchiectasis, Bronchial asthma

Dittrich' s plugs

SPUTUM - MACROSCOPIC STRUCTURES - Hard concretions in a bronchus (lung stones) - CLIN SIG: Histoplasmosis (most common) ; Chronic tuberculosis

Pneumoliths or Broncholiths

SPUTUM - MACRO and MICRO STRUCTURES seen in BRONCHIAL ASTHMA?

"3C-D" -charcot leyden -curshmann's spiral -creola bodies - dittrich's plugs

SPUTUM - MICROSCOPIC STRUCTURES Colorless, hexagonal, double pyramid, often needle-like; arise from disintegration of eosinophils Bronchial asthma (3 C’s), AMOEBIASIS

CharcotLeyden crystals

SPUTUM - MICROSCOPIC STRUCTURES Heart failure cells: hemosiderin-laden macrophage ; Carbon-laden cells: angular black granules Congestive heart failure ; Heavy smokers

Pigmented cells

SPUTUM - MICROSCOPIC STRUCTURES Coiled mucus strands; Can also be observed macroscopically Bronchial asthma (3 C’s)

Curschmann’s spirals

SPUTUM - MICROSCOPIC STRUCTURES Colorless globules occurring in a variety of sizes and bizarre forms No significance; Mistaken as Blastomyces

Myelin globules

SPUTUM - MICROSCOPIC STRUCTURES Clusters of columnar epithelial cells/ KUMPOL KUMPOL Bronchial asthma (3 C’s)

Creola bodies

- A procedure for collecting the cellular milieu of the alveoli by use of a bronchoscope through which saline is instilled into distal bronchi and then withdrawn - Important diagnostic test for Pneumocystis carinii (P. jirovecii) in immunocompromised patients

BRONCHOALVEOLAR LAVAGE (BAL)

best STAIN delineates the cysts of Pneumocystis jirovecii

Grocott's methenamine silver stain

CELLS SEEN IN BRONCHOALVEOLAR LAVAGE Interstitial disease, pulmonary lymphoma, nonbacterial infections

1-15% Lymphocytes

CELLS SEEN IN BRONCHOALVEOLAR LAVAGE Cigarette Smokers, bronchopneumonia, toxin exposure

<3% Neutrophils PRIMARY GRANULOCYTES

- Used to diagnose Cystic fibrosis (Mucoviscidosis) o Autosomal recessive metabolic disorder affecting the mucous secreting glands of the body o Associated with pancreatic insufficiency, respiratory distress & intestinal obstruction o ↑ Na+ & Cl- due to inability of the sweat glands to reabsorb them before the sweat is secreted

SWEAT TEST

- Pilocarpine + mild current = induce sweat production - Application of 0.16 mA current for 5 minutes

GIBSON AND COOKE PILOCARPINE IONTOPHORESIS

Sweat Na+ and Cl- values: - _______= Diagnostic for CF - ______ = Borderline for CF (Repeat testing)

- >70 mEq/L= Diagnostic for CF - 40 mEq/L= Borderline for CF (Repeat testing)

CEREBROSPINAL FLUID FUNCTIONS?

- 3rd major body fluid - Functions: "NMB" o Supply nutrients to the nervous system o Remove metabolic waste o Produce a mechanical barrier to cushion the brain & Spinal cord against trauma

CSF Layer of meninges that Lines the skull & vertebral canal

Dura mater (Outer layer)

CSF Layer of meninges; Portion where CSF flows

Subarachnoid space (Below arachnoid)

CSF Layer of meninges ; Filamentous inner membrane

Arachnoid mater (Spiderweb-like)

CSF Layer of meninges ;Lines the surface of brain & spinal cord

Pia mater (Innermost layer)

CSF produces CSF by selective filtration (at a rate of 20 mL/hour) --- USING MANOMETER

CHOROID PLEXUS

CSF Reabsorbs CSF

ARACHNOID VILLI/GRANULATIONS

CSF o Protects brain from chemicals & other substances circulating in the blood that can harm the brain tissue o Disruption of BBB allows WBCs, proteins & other chemicals to enter the CSF (Ex: Meningitis, Multiple sclerosis)

BLOOD BRAIN BARRIER (BBB)

CSF - Up to ___mL CSF can be collected using a manometer attached to a spinal needle - Method of collection = ________ (between L3-L4 [adults] or L4-L5 [infants])

-20mL - LUMBAR PUNCTURE

CSF Tube 1? Tube 2? Tube 3? (Tube 4)?

Tube 1 = Chemistry/Serology Freezing temperature -- > -20C Tube 2 = Microbiology Room temperature - 25-27C for 30 mins (CM books); 35-37C (Microbiology books) Tube 3 = Hematology Refrigeration temperature/2-6C for 4 hrs (Tube 4) Microbiology or Serology

CSF TOTAL VOLUME - Adults = - Neonates =

- Adults = 90-150mL - Neonates = 10-60 mL

CSF If 1 CSF tube only? what is the order of sections?

"MHC" Microbiology--- Hema -- Chem/Serology

CSF - Crystal clear - Xanthochromic (Pink/Yellow/Orange)

-Normal - Pink = Slight amount of oxyhemoglobin Yellow = Oxyhemoglobin → Bilirubin Orange = Heavy hemolysis -- Other causes: ↑ Carotene, ↑ Melanin, ↑ Protein (> 150 mg/dL), Rifampin

CSF Traumatic Tap Distribution of blood on 3 tubes: Clot formation: Supernatant: Erythrophages (Macrophages w/ ingested RBCs): D-dimer:

Uneven (1>2>3) (+) Due to plasma fibrinogen CLR ABSENT NEG

CSF Intracranial Hemorrhage Distribution of blood on 3 tubes: Clot formation: Supernatant: Erythrophages (Macrophages w/ ingested RBCs): D-dimer:

-Even (1=2=3) -(-) CSF has no fibrinogen -Xanthochromic (RBCs in CSF lyse after 2 hours) -Present (+) Hematoidin & Hemosiderin -Positive

CSF Formation of Pellicle

Tubercular meningitis

CSF Normal values: WBC o Adults = o Neonates =

o Adults = 0-5 WBCs/uL o Neonates = 0-30 WBCs/uL

CSF Clear Slightly Hazy Hazy Slightly Cloudy Cloudy/Slightly Bloody Bloody/Turbid

Undiluted 1:10 1:20 1:100 1:200 1:10,000

CSF RBC Count - Done only in cases of traumatic tap - To correct for WBC count & total protein concentration o -1 WBC for every _____ rbc? o -8 mg/dL total protein concentration for every __ ? o -1 mg/dL total protein concentration for every __?

- 700 RBCS seen - 10,000 RBCs/uL -1,200 RBCs/uL

CSF IN CYTOCENTRIFUGE, WHAT IS ADDED TO Increases cell yield or recovery, Decreases cellular distortion?

Addition of 30% albumin

PREDOMINANT CELLS IN CSF - Adults? - Neonates?

PREDOMINANT CELLS IN CSF - Adults: (70:30 ratio) o 70% Lymphocytes o 30% Monocytes - Neonates (inversed ratio) O Up to 80% MONOCYTES (considered normal) O 20% Lymphocyte

CSF TERM USED? o an abnormal increase in the number of white blood cells in the cerebrospinal fluid (CSF)

PLEOCYTOSIS

-CSF - PROTEIN IS INCREASED IN WHAT CONDITIONS? - Major CSF protein? 2nd Most Prevalent? Alpha-globulins? Beta-globulins? Gamma-globulins? NOT found in normal CSF?

-meningitis, - multiple sclerosis - hemorrhage -Albumin -Prealbumin -ceruloplasmin, haptoglobulin, a2 macroglobulin "CHA" -Beta2-transferrin (tau) o Carbohydrate-deficient transferrin o Found in CSF but not in serum -IgG and some lgA -IgM, Fibrinogen, Lipids

CSF - PROTEIN Normal values - Adults = - Infants = - Immature =

- Adults = 15-45 mg/dL (<1% or 1/200 that of serum protein) - Infants = 150 mg/dL - Immature = 500 mg/dL

CSF PROTEIN DETERMINATION - Precipitates albumin only - To precipitate globulins, add sodium sulfate (Na2SO4

3% Sulfosalicylic acid (SSA) - TURBIDIMETRIC

CSF PROTEIN DETERMINATION - Preferred method - Precipitates BOTH albumin & globulins

3% Trichloroacetic acid (TCA) - TURBIDIMETRIC

Oligoclonal Banding in CSF but NOT in SERUM

(Ms. Neng) - Multiple Sclerosis (persistent) - Neurosyphilis - Encephalitis - Neoplastic disorders - Guillain-Barré syndrome

CSF PROTEIN DETERMINATION - Protein binds to dye → Dye turns from red to blue - ↑ Protein = ↑ Blue color

Coomassie Brilliant Blue

CSF ELECTROPHORESIS -These bands indicate immunoglobulin production - Done in conjunction with serum electrophoresis to ensure that banding is due to neurologic inflammation

oligoclonal bands

Oligoclonal Banding in SERUM but NOT in CSF

- Leukemia - Lymphoma - Viral infections -------Bands may also appear in CSF as a result of BBB leakage or traumatic tap

- Protein component of the lipid-protein complex that insulate the nerve fibers - Presence of MBP in CSF indicates destruction of myelin sheath - Used to monitor the course of multiple sclerosis -MTD : IMMUNOASSAY

MYELIN BASIC PROTEIN (MBP

- Demyelinating disorder - Findings: o (+) Anti-myelin sheath autoantibody o (+) Oligoclonal band in CSF but not in serum o (+) Myelin basic protein (MBP) o ↑ IgG index AND PARESTHESIA

MULTIPLE SCLEROSIS

-Done in conjunction with blood glucose -Specimen for blood glucose should be drawn 2 hours prior to spinal tap (to allow time for equilibration between CSF and plasma glucose)

CSF GLUCOSE (GLU

Oligoclonal Bandingin Serum AND CSF

- HIV

CSF GLUCOSE (GLU NORMAL VALUE?

-- 60-70% of blood glucose (65% or 2/3) - (50-80 mg/dL)

- Inversely proportional to glucose - NV : 10-22 mg/dL

CSF LACTATE (LAT)

- Product of ammonia & alpha-ketoglutarate - Indirect test for the presence of excess ammonia in the CSF - INCREASED IN Reye's syndrome

CSF GLUTAMINE

NV OF CSF GLUTAMINE?

Normal value - 8-18 mg/dL

CSF ENZYMES - ↑ in stroke, MS, degenerative disorders, brain tumors, viral & bacterial meningitis, seizures -↑ in intracerebral and subarachnoid hemorrhage, bacterial meningitis

- CK - AST

CSF ENZYMES Serum LDH: -Normal Serum LDH? -Flipped pattern ? CSF LDH: o Normal pattern = o Neurological abnormalities = o Bacterial meningitis = CSF LDH ISOENZYMES LD 1 & 2: LD 2 & 3: LD 4 & 5:

Serum LDH: o Normal = 2>1>3> 4>5 o Flipped pattern (AMI) = 1>2 CSF LDH: o Normal pattern = 1>2>3>4>5 o Neurological abnormalities = 2>1 o Bacterial meningitis = 5>4>3>2>1 (↑Neutrophils) CSF LDH ISOENZYMES LD 1 & 2: Brain tissues LD 2 & 3: Lymphocytes LD 4 & 5: Neutrophils

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS BACTERIAL MENINGITIS -↑ WBC Type: -PROTEIN: -LACTATE: -GLUCOSE: -AGENTS:

BACTERIAL MENINGITIS -↑ WBC Type: Neutro -PROTEIN: ↑↑ -LACTATE: ↑↑ -GLUCOSE: ↓↓ -AGENTS: "in Senior High, Not Super Landi" S. agalactiae --------- birth to 1m old H. influenzae --------- 1m old to 5 yrs old N. meningitidis ------ 5 yrs old to 29 yrs old Strep. pneumoniae-- >29 yrs old L. monocytogenes-- infants, elderly, immunocom ------ (+) Gram stain - (+) Culture - positive Limulus Lysate test

SEROLOGIC and MOLECULAR TESTING = for detection of bacterial antigens? = recommended by CDC for the detection of Neurosyphilis? = amplification of regions of rRNA genes to detect differentiate agents of meningitis?

=Latex agglutination test and ELISA =VDRL = PCR

CSF -Detects Gram-negative endotoxin in body fluids & surgical instrument Reagent: ?

LIMULUS AMOEBOCYTE LYSATE (LAL) TEST RGT: Blood of horseshoe crab (Limulus polyphemus); Hemocyanin = copper ---Principle o In the presence of endotoxin, the amoebocytes (WBCs) will release lysate (protein) o (+) Clumping or clot formation

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS VIRAL MENINGITIS -↑ WBC Type: -PROTEIN: -LACTATE: -GLUCOSE: -AGENTS:

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS VIRAL MENINGITIS -↑ WBC Type: Lymphocytes -PROTEIN: ↑ -LACTATE: N -GLUCOSE: N -AGENTS: Enteroviruses Poliovirus Echovirus Coxsackievirus

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS TB MENINGITIS -↑ WBC Type: -PROTEIN: -LACTATE: -GLUCOSE: -AGENTS:

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS TB MENINGITIS -↑ WBC Type: Lymphocytes , Monocytes -PROTEIN: ↑↑ -LACTATE: ↑↑ -GLUCOSE: ↓↓ -AGENTS: Mycobacterium tuberculosis ---- (+) AFB stain - (+) Pellicle or web-like clot formation after 12-24 hour refrigeration

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS FUNGAL MENINGITIS -↑ WBC Type: -PROTEIN: -LACTATE: -GLUCOSE: -AGENTS:

CSF - DIFFERENTIAL DIAGNOSIS OF MENINGITIS FUNGAL MENINGITIS -↑ WBC Type: Lymphocytes , Monocytes -PROTEIN: ↑↑ -LACTATE: ↑↑ -GLUCOSE: ↓↓ -AGENTS: Cryptococcus neoformans --- (+) Gram stain = classic starburst pattern - (+) Latex agglutination test - (+) India ink stain = capsule (unstained); background (black) - Cryptococcal meningitis is now commonly encountered in the clinical laboratory

BODY FLUIDS Flashcards

(110 cards)