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GIARDIA LAMBLIA TEST - detection from stool

Giardia Lamblia Antigen


Parasitosis is still a serious global health problem nowadays. 

Giardia Lamblia is the most common protozoan responsible for a wide range of conditions, ranging from severe diarrhea, especially in immunocompromised patients. 

Attachment of the parasite to the duodenal mucosa leads to malabsorption. Intestinal villi atrophy and disappear, leading to significant changes in digestive function, with weight loss and dehydration. In most cases the infection is asymptomatic. 


The Giardia Lamblia quick test on the cassette is a rapid test for immunochromatographic identification and qualitative detection of Giardia Lamblia antigens - 65 kDa coproantigen - a glycoprotein present in Giardia Lamblia cysts and trophozoites. 

Below is presented the work procedure available for most of the quick tests using the immunochromatographic qualitative method of detection.  














IONOMETRY

ION: atom/group of atoms with electric charge (positive charge: cations; negative charge: anions). Many normal substances exist in the body as ions (sodium, potassium, calcium, chloride, bicarbonate, substances known as electrolytes). Chemically, electrolytes are substances that become ions in solution and acquire the capacity to conduct electricity (ions enable the flow of electrical signals through the body). 

The balance of the electrolytes in our bodies is essential for normal function of our cells and organs. If electrolyte levels are too low or too high, cell and organ functions will decline, which could lead to life-threatening conditions. Virtually every metabolic process depends on the presence of electrolytes, which create an "electric potential" needed to do "cellular work".  


The body has complex systems for monitoring and maintaining electrolyte concentrations in a normal narrow range. Physiological functions: 

- maintenance of osmotic pressure and water distribution (water homeostasis)

- conduction of neuromuscular impulses

- acid-base maintenance

- enzyme activation

- electron transfer

- rebuild damaged tissue


Common electrolytes that are measured by doctors with blood testing include sodium, potassium, chloride, bicarbonate, calcium, magnesium, phosphate. These have essential functions in the body. For example, a muscle needs calcium, sodium and potassium to contract. When these substances become imbalanced, it can lead to either muscle weakness or excessive contraction. The heart, muscle and nerve cells use electrolytes to carry electrical impulses to other cells.

Sodium, chloride, potassium, calcium and magnesium (the main electrolytes; nutritional elements) are minerals, and when minerals dissolve in water, they separate into positive and negative ions. For example, when sodium chloride (NaCl) is dissolved in water, it separates into positive sodium ions and negative chloride ions.

Anions: negatively charged ions; migrate towards the anode (positive electrode).

Cations: positively charged ions; migrate towards the cathode (negative electrode).


Major electrolytes (Na+, K+, Cl-, HCO3-) exist as free ions in solution, capable of carrying an electrical charge. Often classified as: major intracellular anion/cation, major extracellular anion/cation. 

Much of the metabolic energy produced by the body is used to establish high intracellular concentrations of potassium (K+) and low concentrations of sodium (Na+), the reverse of the relative concentrations of these ions in the extracellular fluids. The extrusion of sodium requires its movement against a gradient of concentration (higher outside than inside) and electrical potential (inside about 70 mV more negative than outside); work is therefore needed to overcome this electrochemical gradient. 



Functions of the main electrolytes (ions):


- sodium ions: regulate osmotic pressure and the body's water content, transmit nerve signals, contract muscles etc;

- potassium ions: transmit nerve signals and contract muscles including the heart etc;  

- magnesium ions: contract muscles, form bones and teeth, activate enzymes etc;

- calcium ions: transmit nerve signals, contract muscles, form bones and teeth, clot blood;

- chloride ions: regulate osmotic pressure and the body's water content, enable the secretion of stomach acid etc. 

Values differ slightly between serum fluid and plasma fluid; difference is most significant with potassium (serum K+ > plasma K+).

An ion-selective analyser comprises of ion-selective electrode (ISE) and is an analytical technique used to determine the activity of ions in aqueous solution by measuring the electrical potential. It has application in the field of pharmaceuticals and biotechnology. There are four types of ISE: glass electrode, crystalline electrodes, ion-exchange electrodes and enzyme electrodes.


Sodium and potassium and the main and most basic ions analysed. Therefore, more emphasis will be placed on them.  



SODIUM:


Sodium is the major positive ion (cation) in fluid outside of cells. Many processes in the body, especially in the brain, nervous system and muscles require electrical signals for communication. The movement of sodium is critical in the generation of these electrical signals. 


Regulation of osmolality: 

- aldosterone: stimulates sodium (and H2O) retention by kidney (at expense of K+).


Sodium (Na+):

- major cation in ECF (plasma);

- responsible for almost 1/2 the osmotic strength of plasma.


Sodium (Na+) Regulation:

- kidneys are the primary regulators of body sodium and water;

- sodium is freely filtered by glomerulus, 70-80% actively reabsorbed by PCT;

- in times of deficit all sodium is reabsorbed by kidney. 



POTASSIUM:


Potassium is the major positive ion (cation) found inside of cells. Among the many functions of potassium in the body are regulation of the heartbeat and the function of the muscles. 

Potassium levels often change with sodium levels. When sodium levels go up, potassium levels go down, and when sodium levels go down, potassium levels go up. Potassium levels are also affected by a hormone called aldosterone, which is made by the adrenal glands.   

Potassium levels can be affected by how the kidneys are working, the blood pH, the amount of potassium you eat, the hormone levels in your body, severe vomiting, and taking certain medicines, such as diuretics and potassium supplements. Certain cancer treatments that destroy cancer cells can also make potassium levels high. 


Potassium (K+):

- major intracellular cation;

- neuromuscular excitability, heart contractions, maintain ICF volume, maintain H+ concentrations;

- 23x higher in RBC compared to plasma; higher in tissue cells.


Potassium (K+) Regulation:

- Na+-K+-ATP-ase pump continually transports K+ into cells against concentration gradient;

- high intracellular "stores" aid to maintain near-normal extracellular K+ levels in times of deficit;

 - kidney response to conserve K+ is not as immediate and thorough as its response to conserve Na+;

- aldosterone: when sodium is retained, K+ (or H+) is excreted into urine.



CHLORIDE:


Chloride is the major anion (negatively charged ion) found in the fluid outside of cells and in the blood. Chloride plays a role in helping the body maintain a normal balance of fluids. 


Chloride (Cl-):

- major anion in ECF;

- most often Cl- ions shift with Na+ (passive association) and HCO3- to maintain electrical neutrality, osmolality and blood volume. 


Chloride (Cl-) Regulation:

- filtered by glomerulus, passively reabsorbed by the proximal tubules and actively reabsorbed by the ascending Loop of Henle;

- chloride shift: maintains anion-cation balance during buffering process in response to cellular metabolism.



BICARBONATE:


The bicarbonate ion acts as a buffer to maintain the normal levels of acidity (pH) in blood and other fluids in the body. Bicarbonate levels are measured to monitor the acidity of the blood and body fluids.  


Bicarbonate (HCO3-):

- 2nd most abundant anion in ECF;

- maintenance of acid-base balance as the major buffer ion in the carbonic acid/bicarbonate buffer system; major component (>95%) of total CO2 (tCO2).


Bicarbonate (HCO3-) Regulation:

- filtered by the glomerulus and reabsorbed in the proximal and distal tubules;

- kidney has the capacity to reabsorb all or none of the filtered HCO3- as needed to maintain acid/base balance in body.

SODIUM

Sodium: serum and urinary


Serum sodium

All the movements of sodium produce the movement of a variable amount of water. The volume of fluid in the extracellular compartment is directly dependent on the total amount of sodium in the body. The plasma sodium concentration is identical to that of the interstitial fluid. 

In its movements, to achieve an electrical balance, Na+ is followed by anions and primarily by Cl- and HCO3-. Being an alkaline metal and in its movement entraining the bicarbonate anion, Na+ intervenes in maintaining the acid-base balance. Also, Na+ intervenes in the neuromuscular excitability and in the dynamics of the polarization and depolarization phenomena of the cell membrane, opposing the effects of potassium. 

The mechanisms by which the body maintains constant sodium levels in the plasma and extracellular sector are represented by: renal blood flow carbonic anhydrase activity, renin-angiotensin-aldosterone system, ADH, vasopressin, other steroid hormones whose plasma concentration is controlled by he anterior pituitary. 

Biological reference range: premature babies: 132-140 mEq/L; newborns: 133-142 mEq/L; children (1-16): 136-145 mEq/L; adults: 136-145 mEq/L. 


Urinary sodium

The main way to eliminate sodium is through the kidneys. Sodium excretion by the kidneys is influenced by changes in glomerular filtration rate, serum sodium concentration, adrenal cortex activity, the amount of non-resorbable solvates in the filtrate and the volume of extracellular fluid. 

Biological reference range: children: 41-115 mEq/24h; adults: 40-220 mEq/24h.

POTASSIUM

Potassium: serum and urinary.


Serum potassium

Potassium is the main electrolyte (cation) and constituent of the intracellular fluid buffer system. 90% of the potassium is concentrated inside the cell, only small amounts being present in the bones and blood. The damaged cells release potassium into the blood. The entire amount of potassium contained in ingested food is absorbed in the small intestine. The vast majority of potassium (90%) is in ionic form, the rest being protein-related. 

Potassium is indispensable for the normal development of membrane electrical phenomena. It also plays an important role in nerve conduction, muscle contraction, acid-base balance, osmotic pressure, protein anabolism and glycogen formation. Anabolic processes are accompanied by the fixation of potassium in the cell, and catabolic processes by its release. 

Along with calcium and magnesium, potassium controls heart contraction and flow. Potassium and sodium ions are important in the renal regulation of acid-base balance, hydrogen ions being replaced by sodium and potassium ions in the renal tube. Potassium bicarbonate is the major intracellular inorganic buffer. 

Biological reference range:

Premature - umbilical cord blood: 5.0-10.2 mEq/L; premature 48h - venous blood: 3.0-6.0 mEq/L; newborns - umbilical cord blood: 5.6-12.0 mEq/L; newborns - venous blood: 3.7-5.9 mEq/L; children - venous blood: 3.4-4.7 mEq/L; adults: 3.5-5.1 mEq/L. 


Urinary potassium

The greatest amount of potassium is eliminated from the body through the kidneys. Normally 80-90% of potassium is excreted in the urine and the rest in sweat and stool. At the level of the nephron, potassium is subjected to the three fundamental mechanisms: glomerular filtration, complete reabsorption in the proximal tubules and secretion in the distal tubules. 

Biological reference range:

Girls (6-9y): 17-54 mEq/24h; boys (6-9y): 8-37 mEq/24h; girls (10-14y): 22-57 mEq/24h; boys (10-14y): 18-58 mEq/24h; adults: 25-125 mEq/24h.

PHOSPHOROUS

Phosphorous: serum and urinary.


Serum phosphorous

Phosphorous is, after calcium, the most abundant mineral element in the body, being in any tissue. Phosphate is the major intracellular anion. In the cell, phosphorous is mainly involved as organic phosphorous in carbohydrate and lipid metabolism or is bound to proteins and only a small part is present as a phosphate ion. The kidneys are the main regulators of phosphorous homeostasis; approximately 80% of the glomerular filtered amount is reabsorbed in the proximal tubules and 10% in the distal tubules. 

Biological reference range:

Newborns: 4.5-9.0 mg/dL; children: 4.5-5.5 mg/dL; adults: 2.7-4.5 mg/dL. 


Urinary phosphorous

It is recommended to determine urinary phosphorous for the evaluation of phospho-calcium balance and nephrolithiasis. 

Biological reference range: 400-1300 mg/24h; first morning urine: 40-140 mg/dL. 


MAGNESIUM

Magnesium: serum and urinary.


Serum magnesium

Magnesium is an element that, although found in small proportions in the body (0.05% of total body weight), is of great structural and functional importance. About 1% is in plasma, 25% is protein bound, the rest remains in ionized Mg2+ form. In erythrocytes the amount of magnesium is appreciable, about 5.2 mEq/L. Together with Na+, K+ and Ca2+ ions, magnesium regulates neuromuscular excitability and the coagulation mechanisms. The actions of calcium and magnesium are closely linked, the deficiency of one of these elements significantly influencing the metabolism of the other. 

Biological reference range:

Newborns (2 - 4 days old): 1.5-2.2 mg/dL; children (5 months - 6 years old): 1.7-2.3 mg/dL; children (6 - 12 years old): 1.7-2.1 mg/dL; adults: 1.6-2.6 mg/dL. 


Urinary magnesium

Normally 95% of the amount of magnesium that is filtered at the glomerular level is reabsorbed tubularly, especially in the ascending portion of the Henle loop. Magnesium excretion controls the serum level of this element and is dependent on diet. Magnesium, along with calcium, is subject to the effects of parathyroid hormone.  

CALCIUM

Calcium: serum - total and ionic, urinary.


Total serum calcium

Calcium is one of the most important minerals in the body. It is essential for the proper functioning of muscles, nerves, heart, and has an important role in coagulation and bone mineralization. Calcium is the major mineral component of bones. 99% of the body's calcium is in the bones and teeth, which is a huge reservoir for maintaining serum calcium levels, and the rest is distributed in biological fluids and soft tissues. Calcium homeostasis is maintained by parathyroid hormone (PTH). 

About half of the total amount of plasma calcium (45%) is bound to albumin (and only a small portion bound to globulins) in a non-ionized and non-diffusible form, making it a physiological inactive form. A small amount of calcium (5%) is diffusible but not ionized, being represented by citrate, phosphate and calcium bicarbonate. The rest of the plasma calcium is found in ionic or free form and constitutes the physiologically active fraction in the processes of haemostasis and regulation of neuromuscular excitability; its plasma concentration is directly related to PTH and 1,25(OH)2D. 

Calcium in the blood is tested to diagnose and monitor certain diseases related to bones, heart, kidneys and teeth. Its levels do not directly show how much calcium is in the blood, but how much calcium circulates in the blood. Calcium testing can be used if the patient has: kidney stones, bone diseases, neurological disorders.

Abnormal serum calcium levels may indicate parathyroid disfunction, bone disease, carcinoma, malnutrition and malabsorption syndrome, vitamin D deficiency, and kidney disease. Calcium ions play an important role in the transmission of nerve impulses, muscle contraction, heart function and coagulation processes. 

Biological reference range:

0 - 10 days old: 7.6-10.4 mg/dL; 10 days - 3 years old: 6.7-9.8 mg/dL; 3 - 9 years old: 8.4-10.2 mg/dL; 4 - 11 years old: 8.9-10.1 mg/dL; 11 - 13 years old: 8.8 - 10.6 mg/dL; 13 - 15 years old: 9.2-10.7 mg/dL; 15 - 18 years old: 8.4-10.7 mg/dL; adults: 8.8-10.4 mg/dL. 


Serum ionic calcium

Determination of ionic calcium provides guidance on the effect of total protein and albumin on serum calcium levels. A patient may have a high level of total calcium with a normal level of ionic calcium due to an increase in total protein and/or albumin, as is the case with dehydration or multiple myeloma. 

Ionic calcium testing is performed on patients who have a disturbed balance between bound and free calcium, and if they have transfusions, have had major surgery, or have abnormal blood protein levels, such as albumin. High fluctuations in ionic calcium can slow the heart or increase its heart rate, as well as cause muscle spasms, confusion or coma. In the case of very sick patients, it is extremely important to know the level of ionized calcium in order to intervene and prevent major complications. 

Biological reference range:

Newborns: 4.40-5.48 mg/dL; 1-18 years old: 4.80-5.52 mg/dL; adults: 4.65-5.28 mg/dL. 


Urinary calcium

Testing for calcium in the urine shows if the kidneys are excreting the right amount of calcium, and testing for vitamin, phosphorous, and/or magnesium determines what other deficiencies or excesses there are. 

Most of the calcium is excreted in the faeces and a small amount of calcium is excreted in the urine, depending on the calcium intake in the diet. Determination of urinary calcium is important in the diagnosis of hypercalcemia responsible for kidney stones. 

Biological reference range:

Normal diet: 100-300 mg/24h; low calcium diet: 50-150 mg/24h.