Clinical Pathology

Clinical Pathology

Bence Jones Protein

B.J.Protein

    Bence Jones protein is a specialized type of protein found in the urine, specifically associated with certain types of blood cancers known as multiple myeloma or related disorders. Named after the physician who first described them, Henry Bence Jones, these proteins are monoclonal immunoglobulin light chains, either kappa or lambda, produced by abnormal plasma cells in the bone marrow.

Sample:Urine
Check the protein in Salfosasanic acid
Dilution depend on protein value

Protein	Dilution in Distil Water
Trace/1+	1+1
2+	1+2
3+	1+3
4+	1+4

Dilution Example :
Urine protein 4+
1 ml urine sample + 4ml Distil water
2nd tube :
4ml concerntration HCL

Result :- Upper layer white color test is Positive

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BSBP

Bile Salt/ Bile Pigment

Sample :- Urine

Bile Salt

    Bile salts are specialized molecules synthesized in the liver and secreted into bile, a digestive fluid stored in the gallbladder and released into the small intestine during digestion. These amphipathic compounds play a crucial role in the digestion and absorption of fats and fat-soluble vitamins.

3-4 ml urine sample
Add Sulfur powder slowly sprinkle in tube

Result : -
If powder goes down Bile Salt is Present

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Bile Pigment

    Bile pigments are compounds derived from the breakdown of heme, a component of hemoglobin found in red blood cells. These pigments are responsible for the characteristic color of bile and play essential roles in waste excretion and liver function.

3-4 ml urine
Equal amount of Barium Chloride (mix well)
Add 2-3 drops Vouchets reagent
centrifuge 4-5 minute

Result :-
If deposite given Green color Bile Pigment Present.

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Urine for Porphobilinogen

    Porphobilinogen (PBG) is an intermediate compound in the biosynthesis of heme, a vital component of hemoglobin and various enzymes involved in oxygen transport and metabolism. PBG is synthesized in the mitochondria from the condensation of two molecules of δ-aminolevulinic acid (ALA), an initial precursor in the heme biosynthetic pathway. The subsequent conversion of PBG into uroporphyrinogen III through a series of enzymatic reactions ultimately leads to the formation of heme.

2 ml urine
2 ml Enrich reagent
If Dark RedColor

Add sodium Acetate Solution 2 ml mix well
keep it 10 to 15 minutes
If upper layer Reddish/Pinkish color

Take upper laye in another tube
Add equal amount of Butanol mix well
Keep it 10 to 15 minutes

Result : -
If lower layer Reddish/Pinkish color then Porphobilinogen Positive

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Urine for Myoglobuline

    Myoglobin is a protein primarily found in muscle tissues, where it plays a crucial role in oxygen storage and transport within muscle cells. It serves as an oxygen reservoir, releasing oxygen to fuel muscle contractions during periods of increased activity or when oxygen supply is limited. Here's a detailed overview of myoglobin and its significance:

Structure and Function:
Protein Structure: Myoglobin is a globular protein composed of a single polypeptide chain folded into a compact tertiary structure. It contains a heme group, similar to hemoglobin, which binds oxygen reversibly.

Oxygen Binding:
Myoglobin binds oxygen in tissues where oxygen concentration is high, such as the lungs, and releases it when oxygen levels decrease, such as during muscle contraction. This process helps facilitate oxygen delivery to muscles during exercise and contributes to overall muscle function.

Muscle Oxygenation:
Myoglobin enhances the diffusion of oxygen within muscle fibers, allowing for efficient oxygen delivery deep into the muscle tissue, particularly in fast-twitch muscle fibers involved in rapid, intense activities like sprinting or weightlifting.

2 ml urine
2 mg Ammonium Sulphate Crystals
Mix well and centrifuge

Result : -
Upper layer show Reddish color Test is Present

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Occult Blood

    Occult blood refers to the presence of blood in feces that is not visibly apparent to the naked eye. This microscopic bleeding can originate from various parts of the gastrointestinal tract, including the esophagus, stomach, small intestine, colon, or rectum. While occult blood is not visible in stool samples, it can be detected through laboratory testing, typically using fecal occult blood tests (FOBTs) or fecal immunochemical tests (FITs).

Sample : Stool
Benzidine powder
Hydrogen Peroxide
Stool sample
Mix well

Result : -
Change color Green Test is Positive

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Reducing Substances

    Reducing substances are compounds capable of donating electrons to other molecules, thereby reducing them. In clinical laboratory settings, the term "reducing substances" often refers to sugars, particularly glucose, that can be detected in biological samples such as urine.

Sample : Stool
Benedicts reagent 4ml
Stool sample 2-3 drops
Then boiling and observed change color

Blue	Negative	-
Green	Trace	0.5 - 1.0 %
Pale Yellow	+	1.0 - 1.5 %
Dark Yellow	++	1.5 - 2.0 %
Red	+++	> 2.0 %

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Fructose

Sample: Seme
Selwinoff's reagent 1ml
8 - 10 drops Seme sample
Then boiling

Result : -
If color Reddish or Orange Fructose Present

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