Sunday, April 23, 2017

OSMOTIC FRAGILITY TEST

— measure of red cells to take up fluid without lysing
— used to differentiate types of anemia

Factors:

* shape of RBC
* volume, surface of the RBC
* functional state of the RBC

Target Cell
- cell with the largest surface area
- decreased fragility
Spherocyte
- smallest surface area
- increased fragility

Increased Fragility Decreased Fragility

hemolytic anemias splenectomy
hereditary spherocytosis liver disease
sickle cell disease
IDA,
polycythemia vera
thalassemia

Specimen: heparinized venous blood/ defibrillated fibrous blood

tested be within 2 hrs of collection - 6 hrs

PRINCIPLE:

RBC in isotonic — no change in RBC

RBC in hypotonic — cell either ruptures or an equilibrium/ cell bursts

RBC in hypertonic — cell will shrink

1
 Sunday, April 23, 2017

% NaCl — 0.02 x No. of tube

% hemolysis — OD of supernatant

OD of supernatant tube #14

Normal Hemolysis : 0.33%

ph : 7.4

Malaria
Causative Agent:

Genus: Plasmodium / class: Sporozoa/ suborder: Haemosporidia

Four Species:

Plasmodium vivax

Plasmodium malariae

Plasmodium falciparum

Plasmodium ovale

Vector: female Anopheles mosquito

life cycle:

* Sporogony - sexual cycle

Transmission:

2
 Sunday, April 23, 2017
infected Anopheles mosquito - sporozoites invade parenchymal cells of the liver ~
PRE ERYTHROCYTIC DEVELOPMENT — schizont phase — merozoites are formed
— schizonts — trophozoite rings— mature schizonts

Gametocyte: infective to the Anopheles mosquito

P. falciparum, P. ovale, P. vivat — 48 hrs

P. malariae — 72 hrs

PATHOPHYSIOLOGY

Blackwater Fever

—occur in P. falciparum infections

—severe acute intravascular hemolysis

—severe anemia, chills, weakness, fever and vomiting

Resistant to Malaria:

Sickle Cell

G-6-PD

Pyruvate kinase Deficiency

Duffy antigen receptor negativity

Elliptocytosis

Laboratory Diagnosis
—mild anemia
— OFT is increased (hemolysis present/ haptoglobin is decreased)

3
 Sunday, April 23, 2017
— WBC is increased , thrombocytopenia

Treatment: Quinine

4
 Sunday, April 23, 2017

PLATELETS
—deals primarily in HEMOSTASIS

Normal range: 150,000 - 450, 000/uL

<100,000/ uL — abnormally low

30,000- 50,000/uL - bleeding possible

<30, 000- spontaneous bleeding

<5,000/uL - severe spontaneous bleeding

Increased Decreased

PV Purpura

Idiopathic Thrombocytopenia ALL, Pernicious Anemia

CLL Gaucher’s

Platelet Counting

reagents used:

Ammonium oxalate — diluting fluid

Phase Microscopy

* Tocantin’s Method

* Rees - Ecker ( platelet appears BLUE)

- Brilliant Cresol Blue, Formaldehyde, Sodium citrate

5
 Sunday, April 23, 2017

SICKLE CELLS

Solubility Test
Specimen used: EDTA- whole blood/ heparin/ sodium citrate

RBCs + saponin (lysing agent)

Positive: turbid appearance

False + False -

globulins hemoglobin >7g/dL

lipemia

high hemoglobin

HEMOGLOBINOPATHIES

Hemoglobin Electrophoresis

Cellulose Acetate ( ph 8.4 - 8.6)

- primary screening used to detect variant Hb~

- difference in the net charge of the Hb determine its mobility and manifests itself
but the speed of migration to the positive pole

6
 Sunday, April 23, 2017
Stains: Ponceau stain

Quantitative: Densitometer and read 525 nm

Quantitation: hemoglobin F and A2 (Alkali denaturation/ Radial Immunodiffusion

test)

ACID AND ALKALINE GLOBIN CHAIN ERYTHROPHORESIS

- identification of unknown hemoglobins

- 2 mercapotoethanol and urea

Citrate Agar Electrophoresis ( pH 6.0 - 6.2)

- confirm variant hemoglobin variants

- differentiates Hb S from D & G from Hb

- differentiates Hb C from Oarab and CHarlem

specimen: EDTA- whole blood

- if it exceeds 2 mg/dL ( false higher count)

COAGULATION
Intrinsic Factor - 11, 12

Extrinsic Factor - 7,

Common Pathway - 1, 2, 10

Contact - 8, 9, 11, 12, PK, HK

7
 Sunday, April 23, 2017
Fibrinogen - 1, 5, 8, 13

Prothrombin - 2, 8, 9, 10