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MUST TO KNOW IN CLINICAL MICROSCOPY
URINALYSIS
Nephron Basic structural unit of kidney 1M/kidney Urethra F: 3-4 cm M: 20 cm Urine formation (order) Glomerulus → Bowman’s capsule → PCT → Loop of Henle → DCT → CD PCT 65% of reabsorption ADH Regulate H 2 O reabsorption in DCT and CD Urine composition 95 - 97% H 2 O 3 - 5% solids 60g TS in 24 hrs 35g: Organic = Urea (major) 25g: Inorganic = Cl (#1) > Na+^ > K+ Glomerular Filtration Clearance tests Evaluate glomerular filtration
- Urea clearance
- Creatinine clearance = most common
- Inulin clearance = gold standard
- Beta 2 - microglobulin
- Radioisotopes Creatinine clearance Formula: Cc = U x V x 1. P A Normal values: M = 107-139 mL/min F = 87 - 107 mL/min Tubular Reabsorption Tubular Reabsorption 1 st^ function to be affected in renal disease Concentration tests Evaluate tubular reabsorption Fishberg test (Old) Patient is deprived of fluid for 24hrs then measure urine SG (SG ≥ 1.026) Mosenthal test (Old) Compare day and night urine in terms of volume and SG Specific Gravity (New) Influenced by # and density of particles in a solution Osmolarity Influenced by # of particles in a solution Principle: Freezing point depression
- 1 Osm or 1000 mOsm/kg of H 2 O will lower the FP of H 2 O (0’C) by 1.86’C
- FP = Osm Example: Determine Osm in mOsm/kg Temp. = - 0.90’C Solution: 1000 mOsm/kg = _ _x____
- 1.86’C - 0.90’C x = 484 mOsm/kg Tubular Secretion and Renal Blood Flow PAH test p-aminohippuric acid PSP test Phensulfonphthalein test Obsolete, results are hard to interpret Methods of Collection Midstream/Catheterized Urine culture Suprapubic aspiration Anaerobic urine culture
3 glass technique For detection of prostatic infection
- 1st^ portion of voided urine
- Middle portion of voided urine: Serves as control for kidney and bladder infection
- If (+), result for #3 is considered invalid
- Urine after prostatic massage Compare WBC and Bacteria of specimen 1 and 3 Prostatic infection: 1 < 3 (10x) Pediatric specimen Wee bag Drug Specimen Collection Chain of custody: step by step documentation of handling and testing of legal specimen Required amount: 30 - 45 mL Temperature (urine): 32.5-35.7’C (w/in 4 mins) Blueing agent → Toilet bowl (to prevent adulteration) Types of Urine Specimen Occasional/Single/Random Routine Qualitative UA 24 hr 1 st^ voided urine → discarded w/ preservative Ex. 8AM → 8AM 12 hr Ex. 8AM → 8PM Addis count: measure of formed elements in the urine using hemacytometer Afternoon (2PM-4PM) Urobilinogen (alkaline tide) 4 hr Nitrite determination (1st^ morning/4 hr) NO 3 → NO 2 = (+) UTI 1 st^ morning Pregnancy test (hCG) Ideal specimen for routine UA Most concentrated and most acidic = preservation of cells and casts Fasting/2nd^ morning Glucose determination 2 nd^ voided urine after a period of fasting Changes in Unpreserved Urine Decreased Clarity Bacterial multiplication Precipitation of AU/AP Glucose Glycolysis Ketones Volatilization Bilirubin Photooxidation Urobilinogen Oxidized to urobilin RBC/WBC Disintegrate in alkaline urine Increased pH Urea ---(Urease)---> NH 3 Bacteria Multiplication Odor Urea ---(Urease)---> NH 3 Nitrite Bacterial multiplication Differentiate contamination from true infection Contamination: Bacteria True infection: Bacteria and WBCs Preservation Refrigeration 2 - 8’C SG (hydrometer/urinometer) Precipitate AU/AP Formalin Addis count Boric acid Urine culture
Red/Purple/Burgundy red/ purplish red/Portwine Porphyria (Lead poisoning: normal color) Brown/black Methemoglobin (acid urine) Homogentisic acid: Alkaptonuria
- Urine darkens after a period of standing
- (-) Homogentisic acid oxidase Urine Color Changes w/ Commonly Used Drugs Cola-colored Levodopa (Tx: Parkinsonism) Red → Brown (alkaline) Yellow Mepacrine/Atabrine (Tx: Malaria, Giardiasis) Red to brown Metronidazole/Flagyl (Tx: Trichomoniasis, Amoebiasis, Giardiasis) Methyldopa/Aldomet (Antihypertensive) Orange-red (acid) Phenazopyridine/pyridium (Tx: UTI) Bright orange-red (acid) Rifampin (Tx: TB) = all body fluids are red Bright yellow Riboflavin (Multivitamins) Nubecula Faint cloud in urine after a period of standing WBCs, epithelial cells and mucus Bilifuscin (Dipyrrole) Hemoglobin Köln = unstable Red-brown urine Clarity/Transparency/Turbidity Clear Transparent, no visible particulates Hazy Few particulates, print easily seen through urine Cloudy Many particulates, print blurred through urine Turbid Print cannot be seen through urine Milky May precipitate or clot Bacteria Uniform turbidity NOT cleared by acidification or filtration Chyluria Lymph fluid in urine Filariasis Squamous epithelial cells females Radiographic contrast media SG by refractometer (>1.040) Rgt strip: not affected by RCM Vaginal cream Tx: Candida Pseudochyluria Laboratory Correlations in Urine Turbidity Acidic urine AU RCM Alkaline urine AP Carbonates Soluble w/ heat AU Uric acid Soluble w/ dilute acetic acid RBCs AP Carbonates Insoluble in dilute acetic acid WBCs Yeasts Spermatozoa Bacteria Soluble in ether Lipids Lymph fluid Chyle
Specific Gravity SG Density of solution compared w/ density of similar volume of distilled H 2 O at a similar temperature NV = 1.003-1.035 (random) SG <1.003 = not a urine except DI Refractometer (TS meter) Based on refractive index: RI = light velocity in air light velocity in soln Compensated to temperature (15-38’C) Corrections: a. 1g/dL glucose: (-0.004) b. 1g/dL protein: (-0.003) Calibrations: a. Distilled H 2 O = 1. b. 5% NaCl = 1.022 ± 0. c. 9% Sucrose = 1.034 ± 0. Urinometer Requires temperature correction a. 3’C calibration temperature (20’C) = (+0.001) b. 3’C calibration temperature (20’C) = (-0.001) Requires correction for glucose and protein (Rf/U) Rf < U by 0.002 Refractometer reading is lower than the urinometer reading by 0. Urinometer calibration K 2 SO 4 solution: 1L H 2 O + 20.29g K 2 SO 4 SG = 1. Isosthenuria SG = 1.010 (Glomerular filtrate) Hyposthenuria SG < 1. Hypersthenuria SG > 1. Urine Odor Aromatic/Odorless Normal Ammoniacal Urea ---(Urease)---> NH 3 Ex. UTI ( Proteus : urease) Fruity, sweet DM (Ketones) Rotten fish/Galunggong Trimethylaminuria Sweaty feet Isovaleric acidemia Mousy Phenylketonuria Cabbage Methionine malabsorption Caramelized sugar, curry MSUD Bleach Contamination Sulfur Cystine disorder Chemical Examination of Urine Specific Gravity Principle (Rgt Strip) pKa dissociation constant concentration = H+ Indicator: Bromthymol blue = () Blue → Green → Yellow () Other info. Not affected by glucose, protein and RCM Harmonic Oscillation Densitometry Frequency of soundwave entering a solution will change in proportion to the density (SG) of the solution
- Yellow IRIS (Automated): International Remote Imaging System pH Normal Random = 4.5-8. 1 st^ morning = 5.0-6. pH 9.0 = Unpreserved urine
CSF protein = frequently tested
- Det: TCA (preferred) and SSA SSA Reactions (Protein) Negative No increase in turbidity < 6 mg/dL Trace Distinct turbidity 6 - 30 mg/dL 1+ Noticeable turbidity w/ no granulation 30 - 100 mg/dL 2+ Turbidity w/ granulation but no flocculation 100 - 200 mg/dL 3+ Turbidity w/ granulation and flocculation 200 - 400 mg/dL 4+ Clumps of protein > 400 mg/dL Glucose Glucose Most frequently tested in urine Threshold substance Renal threshold = 160- 180 mg/dL
- Plasma concentration of a substance at w/c tubular reabsorption stops and amount of substance in the urine Other substances in urine ID: TLC
- Fructose (Levulose): fruits, honey syrup
- Galactose: infants (Galactosemia: enzyme deficiencies)
- Galactose- 1 - uridyltransferase deficiency
- Galactokinase deficiency
- Lactose
- During lactation
- Towards the end of pregnancy
- Patient on strict milk diet
- (+) Rubner’s test (Lead acetate)
- Pentose
- Xylose, arabinose
- Xylulose:Benign pentosuria
- Sucrose
- Intestinal disorders
- Nonreducing sugar
- (-) Copper reduction test Hyperglycemia associated Glycosuria Blood glucose, Urine glucose
- DM
- Cushing’s syndrome/disease = cortisol
- Pheochromocytoma = catecholamines
- Acromegaly = GH
- Hyperthyroidism = T3/T Renal associated Glycosuria N-Blood glucose, impaired tubular reabsorption of glucose
- Fanconi’s syndrome: defective tubular reabsorption of glucose and amino acids Principle (Rgt Strip) Double sequential enzyme reaction:
- Glucose oxidase
- Peroxidase Chromogen:
- KI (Brown)
- Tetramethylbenzidine (Blue) Copper Reduction test (Clinitest) Blue tablet Relies on the ability of glucose and other substances to reduce CuSO 4 to Cu 2 O in the presence of alkali and heat CuSO 4 (Blue) -------------> Cu 2 O (Brick red)
Pass through phenomenon Occurs if >2 g/dL sugar is present in urine Blue → Green → Yellow → Brick red → Blue (Pass through) To prevent, use 2 gtts urine (instead of 5 gtts) + 10 gtts H 2 O + Clinitest (-) Glucose oxidase (+) Clinitest (+) Nonglucose reducing substance 1+ Glucose oxidase (-) Clinitest True glucosuria Small amount of glucose present 4+ Glucose oxidase (-) Clinitest False (+) Possible oxidizing agent interference on reagent strip Ketones Ketones Result from increased fat metabolism due to inability to metabolize CHO 78% BHA = major ketone but not detected 20% AA/Diacetic acid = parent ketone 2% Acetone Significance Diabetic acidosis Insulin dosage monitoring Starvation Vomiting Principle (Rgt Strip) Legal’s test (Sodium nitroprusside reaction) AAA + Sodium nitroprusside --------------> (+) Purple (Acetone) (Glycine) Acetest Sodium nitroprusside Glycine Disodium phosphate Lactose Blood Hematuria Cloudy red urine (Intact RBCs) Renal calculi GN Strenuous exercise Anticoagulants Hemoglobinuria Clear red urine Intravascular hemolysis Myoglobinuria Clear red urine Rhabdomyolysis Hgb vs. Mgb 1. Plasma examination
- Hgb: Red/pink plasma, haptoglobin
- Mgb: Pale yellow, CK, Aldolase
- Blondheim’s test (Ammonium SO 4 ): Precipitates Hgb Urine + 2.8g NH 4 SO 4 (80% Satd.) ---(Filter/Centrifuge)---> Supernatant Supernatant: Red = Myoglobin = (+) Rgt strip Clear w/ red ppt. = Hemoglobin = (-) Rgt strip Hemolytic anemia 1 hr post transfusion urine = Hgb Week after = Hemosiderin Principle (Rgt Strip) Pseudoperoxidase activity of hemoglobin Chromogen: TMB [(-) Yellow/(+){}Green → Blue {}] H 2 O 2 + Chromogen ---(Heme)---> Oxidized chromogen + H 2 O Hgb/Mgb Uniform green/blue Hematuria Speckled/spotted Extravascular lysis Unconjugated bilirubin Urine and fecal urobilinogen
Principle (Rgt strip) Indoxyl carbonic acid ester + Diazonium salt ---(LE)---> Indoxyl + Acid indoxyl ----------> (+) Purple Strip can detect even lysed WBCs Reading Time (Reagent Strips) 30 seconds Glucose Bilirubin 40 seconds Ketones 45 seconds SG 60 seconds “PPBUN” pH Protein Blood Urobilinogen Nitrite 120 seconds Leukocytes Vitamin C (Ascorbic acid) 11 th^ reagent pad Reducing property False (-) rgt strip: “BB LNG”
- Blood
- Bilirubin
- Leukocytes
- Nitrite
- Glucose Rgt: Phosphomolybdate Phosphomolybdate + Vitamin C (≥5 mg/dL) --------> (+) Molybdenum blue Sources of Error/Interference (Reagent Strips) False-positive False-negative SG High concentrations of protein Highly alkaline urines (>6.5) pH No known interfering substance Runover from adjacent pads Old specimens Protein Highly buffered alkaline urine Pigmented specimens, phenazopyridine Quarternary ammonium compounds (detergents) Antiseptics, chlorhexidine Loss of buffer from prolonged exposure of the reagent strip to the specimen High specific gravity Proteins other than albumin Glucose Contamination by oxidizing agents and detergents High levels of ascorbic acid High levels of ketones High specific gravity Low temperatures Improperly preserved specimens Ketones Phthalein dyes Highly pigmented red urine Levodopa Medications containing free sulfhydryl grps Improperly preserved specimens
False-positive False-negative Blood Strong oxidizing agents Bacterial peroxidases Menstrual contamination High specific gravity/crenated cells Formalin Captopril High concentration of nitrite Ascorbic acid >25 mg/dL Unmixed specimens Bilirubin Highly pigmented urines, phenazopyridine Indican (intestinal disorders) Metabolites of Lodine Specimen exposure to light Ascorbic acid >25 mg/dL High concentrations of nitrite Urobilinogen Porphobilinogen Indican p-aminosalicylic acid Sulfonamides Methyldopa Procaine Chlorpromazine Highly pigmented urine Old specimens Preservation in formalin Nitrite Improperly preserved specimens Highly pigmented urine Nonreductase-containing bacteria Insufficient contact time between bacteria and nitrate Lack of urinary nitrate Large quantities of bacteria converting nitrite to nitrogen High concentrations of ascorbic acid High specific gravity Leukocytes Strong oxidizing agents Highly pigmented urine, nitrofurantoin High concentrations of protein, glucose, oxalic acid, ascorbic acid, gentamicin, cephalosporins, tetracyclines Microscopic Exam of Urine Phase-contrast microscopy Visualization of elements w/ low refractive indices:
- Hyaline casts
- Mixed cellular casts
- Mucous threads
- Trichomonas Polarizing microscopy ID of cholesterol in OFB, FC and crystals Interference contrast microscopy Produces 3D microscopy-image and layer-by-layer imaging of a specimen
- Hoffman microscope: modulation contrast microscope
- Nomarski microscope: differential interference contrast microscope Sternheimer-Malbin Crystal violet and safranin Nucleus and cytoplasm ID: WBCs, ECs, casts Toluidine blue (Supravital) Enhances nuclear detail Differentiates WBCs and RTE Lipid stains: ORO and Sudan III Stain TG and neutral fats orange red ID: free fat droplets and lipid-containing cells and casts Gram stain Differentiates Gram (+) and Gram (-) bacteria ID: bacterial casts Hansel stain Eosin Y and Methylene blue ID: Eosinophils Prussian blue stain Stains structures containing iron
Pingpong disease S. haematobium “Hematuria” Specimen: 24 hr unpreserved urine E. vermicularis Most common fecal contaminant Casts (Cylindruria) Formed in the DCT and CD ♫ Tamm-Horsfall protein (Uromodulin)
- Major constituent
- Glycoprotein secreted by RTE cells of DCT and CD Hyaline casts NV = 0-2/lpf Beginning of all types of casts (prototype cast) a. Physiologic:
- Strenuous exercise (HC, GC, RC)
- Heat b. Pathologic:
- GN
- PN
- CHF RBC casts Bleeding w/in the nephron a. GN b. Strenuous exercise (HC, GC, RC) WBC casts Inflammation w/in the nephron Differentiates upper UTI (pyelonephritis, w/ cast) from lower UTI (cystitis, no cast) To differentiate from EC cast:
- Phase contrast microscopy
- Supravital stain Seen in:
- PN
- AIN Bacterial casts Pyelonephritis Epithelial cell casts Renal tubular damage Advanced tubular destruction Coarse/Fine granular casts Formed from the disintegration of cellular cast GN PN Strenuous exercise (HC, GC, RC) Fatty casts Nephrotic syndrome: lipiduria Not stained by Sternheimer-Malbin Waxy casts Final degenerative form of all types of casts Stasis of renal flow Chronic renal failure Brittle, highly refractile, w/ jagged ends Broad casts “Renal failure casts” Extreme urine stasis Widening and destruction of tubular walls Any type of cast can be broad Sediment preparation Urine → Centrifuge: 400 RCF for 5 mins → Decant → Remaining: 0.5mL/1.0mL Urine sediment: 20μL (0.02 mL)
- 10 lpf
- 10 hpf
- Reduced light RCF 1.118 x 10-^5 x radius (cm) x (rpm)^2
Urine Crystals Amorphous Urates (Normal) (pH: acid) Yellow-brown granules Pink sediment (Uroerythrin) Mistaken as cystine crystals Rhombic, wedge, rosette, hexagonal, four-sided plate (whetstone) Lemon-shaped (Henry) Lesch-Nyhan syndrome: orange sands in diaper Gout Chemotherapy Calcium Oxalate (Normal) (pH: acid/alkaline/neutral)
- Weddelite = dihydrate
- Whewellite = monohydrate
- Oval, dumbbell
- Ethylene glycol poisoning (antifreeze agent) Most renal stones consist of CaOx Amorphous Phosphates (Normal) (pH: alkaline/neutral) White precipitate Granular appearance After meal (alkaline tide) Ammonium Biurate (Normal) (pH: alkaline) Yellow-brown Thorny apples Old specimen: due to the presence of urea-splitting bacteria Triple Phosphate (Normal) (pH: alkaline) A.k.a. Magnesium ammonium phosphate Coffin lid, “Struvite”, staghorn appearance Presence of urea-splitting bacteria Calcium Phosphate (Normal) (pH: alkaline/neutral) Colorless, flat rectangular plates or thin prisms often in rosette formation Rosettes may resemble sulfonamides
- To differentiate: CaPO 4 dissolves in acetic acid
- Calcium Phosphate = Apatite
- Basic Calcium Phosphate = Hydroxyapatite
- Calcium Hydrogen Phosphate = Brushite Calcium Carbonate (Normal) (pH: alkaline) Small and colorless Dumbbell or spherical shapes Acetic acid: (+) Effervescence Cystine (Abnormal) (pH: acid) Colorless hexagonal plates Cystinuria Cholesterol (Abnormal) (pH: acid) Rectangular plate w/ notch in one or more corners Staircase pattern Lipiduria (Nephrotic syndrome) Resemble crystals of RCM, to differentiate a. Patient history b. Correlate w/ other UA results c. RCM: SG by refractometer ≥1. Tyrosine (Abnormal) (pH: acid/neutral) Colorless to yellow needles Liver disease (more common) (+) Nitroso-naphthol Leucine (Abnormal) (pH: acid/neutral) Yellow-brown spheres w/ concentric circles and radial striations Liver disease Bilirubin (Abnormal) (pH: acid) Clumped needles or granules w/ yellow color (+) Diazo reaction Liver disease
- Oval fat bodies, fatty and waxy casts Telescoped sediments Simultaneous appearance of the elements of acute/chronic GN and nephrotic syndrome Cells and Casts a. Lupus nephritis b. SBE UTI E. coli = 90% cases of UTI S. saprophyticus = UTI among sexually active young females G. vaginalis = bacterial vaginosis S. pyogenes = AGN and ARF Viridans Streptococci = SBE Rapidly progressive (Crescentic) GN Deposition of immune complex from systemic immune disorders on the glomerular membrane Goodpasture syndrome Attachment of cytotoxic antibody to glomerular and alveolar basement membrane Wegener’s granulomatosis Antineutrophilic cytoplasmic autoantibody Henoch-Schönlein purpura Occurse in children following viral respiratory infection Decrease in platelets disrupts vascular integrity Membranous GN Thickening of the glomerular membrane following IgG immune complex deposition Membranoproliferative GN Cellular proliferation affecting the capillary walls or the glomerular basement membrane Chronic GN Marked decrease in renal function resulting from glomerular damage precipitated by other renal disorders IgA nephropathy (Berger’s disease) Deposition of IgA on the glomerular membrane Nephrotic syndrome Disruption of the electrical charges that produce tightly fitting podocyte barrier Minimal change disease (Lipoid nephrosis) Disruption of the podocytes occurring primarily in children following allergic reaction and immunization FSGS Disruption of podocytes in certain areas of glomeruli associated w/ heroin and analgesic abuse and AIDS Alport syndrome Lamellated and thinning of glomerular basement membrane Diabetic Nephropathy (Kimmelstiel-Wilson disease) Most common cause of ESRD Microalbuminuria Acute tubular necrosis Damage to the renal tubules caused by ischemia or toxic agents Fanconi syndrome Generalized defect in renal tubular reabsorption in the PCT Nephrogenic DI Inability of the renal tubules to respond to ADH Neurogenic DI Inability of the hypothalamus to produce ADH Renal glucosuria Inability of the renal tubules to reabsorb glucose Cystitis Ascending bacterial infection of the bladder Acute PN Infection of the renal tubules and interstitium Chronic PN Recurrent infection of the renal tubules and interstitium Visicoureteral reflux: most common cause
- Reflux of urine from the bladder back into the ureters Screening for Metabolic Disorders Aminoaciduria 1. Overflow type AA in blood AA in urine Ex. PKU, alkaptonuria, MSUD
- Renal type N-AA in blood Impaired tubular reabsorption of AA Ex. Cystinuria (COLA), Fanconi’s syndrome Phenylalanine-Tyrosine Disorders Phenylalanine (-) PAH PKU Phenylpyruvic acid Tyrosine Tyrosine transaminase (-) p-Hydroxyphenylpyruvic acid Tyrosinemia Tyrosyluria: p-Hydroxyphenylpyruvic acid oxidase p-OHPPA Homogentisic acid (-) p-OHPLA Homogentisic acid oxidase Alkaptonuria Maleylacetoacetic acid Homogentisic acid Fumarylacetoacetic acid Fumaric acid and Acetoacetic acid Phenylketonuria Severe mental retardation Mousy odor (-) PAH Screen: FeCl 3 → (+) Blue-green Confirm: Guthrie test (Bacterial inhibition)
- B. subtilis
- Inhibitor: Beta 2 - thienylalanine (neutralized by phenylalanine)
- Growth = (+) PKU
- No growth = (-) PKU Tyrosyluria Rancid butter odor (-) Tyrosine transaminase and p-OHPPA oxidase Screen: FeCl 3 → (+) Transient green Confirm: Nitroso-naphthol → (+) Orange-red Alkaptonuria Urine darkens after a period of standing (-) Homogentisic acid oxidase Homogentisic acid in blood and urine FeCl 3 → (+) Transient blue Clinitest/Benedict’s → (+) Yellow ppt. Melanuria Overproliferation of melanocytes FeCl 3 → Gray or black ppt. Ehrlich’s → Red Branched-Chain Amino Acid Disorders MSUD Accumulation of leucine, isoleucine and valine in blood and urine 2,4-DNPH → (+) Yellow turbidity/ppt. Organic acidemias 1. Isovaleric acidemia = sweaty feet
- Propionic acidemia
- Methylmalonic acidemia Tryptophan Disorders Indicanuria Intestinal disorder Blue color
Screening tests (porphyria) 1. Ehrlich reaction = (+) D-ALA and porphobilinogen
- Fluorescence at 550-600nm = Uro/Copro/Protoporphyrin = (+) Red/pink/violet = (-) Blue
- Free Erythrocyte Protoporphyrin (FEP) = CDC recommended test for Lead poisoning Specimens Urine: red/purple/portwine (normal: Lead poisoning) Stool Blood Bile Lead poisoning RBC inclusion coarse basophilic stippling Qualitative Tests for Protein (+) White Ring Heller’s Robert’s Spiegler’s (+) Violet Biuret (Albumin) (+) White turbidity/ cloudiness Heat and acetic acid SSA Purdy’s Potassium ferrocyanide Picric acid Kingsbury-Clark (Rgt: SSA) (+) coagulum (24 hrs) Esbach’s
- Rgt: Picric acid + Citric acid Tsuchiya’s (+) coagulum (72’C for 5mins) Kwilecki’s
- Rgt: Esbach’s + 10% FeCl 3 Qualitative Tests for Sugars Benedict’s Reducing substances Seliwanoff’s Rgt: Resorcinol Fructose → (+) Red Rubner’s Rgt: Lead acetate, NH 3 H 2 O Lactose → (+) Bright red w/ red ppt. Glucose → (+) Red color w/ yellow ppt. Bial Orcinol Pentose → (+) Green Tauber’s Pentose → (+) Green Others Osazone or phenylhydrazine (Kowarsky) Nylander’s Moore Heller Borchardt’s Qualitative Tests for Ketones Frommer’s Acetone → (+) Purplish red ring Rothera’s Acetone & AAA → (+) Purple ring Lange Acetone & AAA → (+) Purple ring Acetest/Ketostix Acetone → (+) Purple Gerhardt’s AAA → Bordeaux red Qualitative Tests for Bile Pigments Gmelin Bile → (+) Play of colors Smith Bile → (+) Emerald green Harrison’s spot Bile → (+) Blue to green
Ictotest Bile → (+) Blue to purple mat Wallace and Diamond Rgt: PDAB Urobilinogen → (+) Cherry red Schlesinger Rgt: Lugol’s iodine, Alc. Zinc acetate Urobilin → (+) Greenish fluorescence Qualitative Tests for Hemoglobin Benzidine (+) Green-blue Guiac (+) Blue Ortho-toluidine (+) Blue Qualitative Tests for Melanin FeCl 3 (Screening) (+) Black (after 24 hrs) Thomahlen (+) Dark green or blue color (fresh urine) Blackberg & Wanger (+) Brown to black ppt. (24 hr urine) Qualitative Tests for Chloride Fantus (+) Reddish ppt Mercurimetric titration (Schales & Schales) (+) Blue-violet colored complex Qualitative Test for Calcium Sulkowitch (+) Precipitation Renal Function Tests Test for Glomerular filtration Clearance Test for Tubular reabsorption Concentration tests
- Fishberg (old)
- Mosenthal (old)
- SG (new)
- Osmolality (new) Fishberg test Patient deprived of fluid for 24 hrs = SG ≥1. Patient deprived of fluid for 12 hrs = SG ≥1. Test for Tubular Secretion and Renal Blood flow
PAH
PSP
Tests for NPN 1. Urea: Urease, DAM (NV = 6-17 g/24 hr urine)
- Creatinine: Jaffe (NV = 1-2 g/24 hr urine)
- Uric acid: Uricase, PTA (NV = 0.25-0.75 g/24 hr urine) BCR (BUN: Crea Ratio) a. NV = 10:
- BUN: 90% excreted, 10% reabsorbed
- Crea: 99% excreted, 1% reabsorbed b. Renal disease: Normal ratio BUN, Crea c. Pre- and Post-renal disease: Ratio BUN, N-crea Other Topics Biohazard Symbol 4 circles Top = Source Left = Host Right = Transmission PPE Gloves Fluid-resistant gowns Eye and face shields Plexiglas countertop shields Disinfection of sink 1:5 or 1:10 dilution of sodium hypochlorite (daily)
