CENBIMO S.L

Lysoyme, also known as Muramidase, catalyses the hydrolysis of some mucopolysaccharides in bacterial cell walls, specifically hydrolyzing between acetylmuramic acid and acetylglucosamine.

This enzyme is found in a variety of human cells but principally in tissular histiocytes, circulating granulocytes, monocytes and in glandular saliva and lacrimal cells.

The quantity of lysozyme produced by monocytes is greatly superior to that of polymorphonuclear neutrophil granulocytes.

Various point mutations in the lysozyme gene have been associated with non-neuropathic visceral amylodosis.

Histosonda Lysozyme consists of a fragment of single stranded DNA 358 nucleotides long that is targeted against lysozyme RNA,

The principle use of this probe is the classification and study of histiocytic cells (especially when they are activated and functioning) and the detection of monocytes in the bone marrow.

Due to the fact that in decalcified, paraffin embedded bone marrow biopsies it is not possible to clearly see the granulocytic cytoplasmic granuoles it is frequently very difficult to distinguish between myelocytes and monocytes.  Histosonda Lysozyme permits a clean and clear differentiation of the bone marrow monocytic cells owing to the augmented transcription of the gene producing lysozyme.  This permits the clear differentiation of chronic myelomonocytic leukemia from other types of chronic granulocytic leukemias.

INTENDED USE

For use in In Vitro Diagnosis.

The Kit Histosonda Lysozyme is useful for the classification and study of histiocytic cells (especially when they are activated and functioning) and the detection of monocytes in the bone marrow.

WARNINGS AND PRECAUTIONS

The Kit Histosonda Lysozyme has been designed for professional use in In Vitro Diagnosis and must be manipulated by qualified and accordingly trained personnel.  In order to obtain the best results, the instructions contained in the manual must be followed. Any change to the indicated temperatures, times or any other step of the process can lead to poor results.

KIT COMPONENTS

The Kit includes 20 single test tubes of Histosonda Lysozyme; 20 single test tubes of Proteinase K and 2 tubes of Anti-Digoxin antibody sufficient for 20 tests in total.  All products are lyophilized.

Histosonda Lysozyme consists of a single stranded DNA fragment with a length of 358 nucleotides targeted against Lysozyme mRNA.  The DNA of this probe has been labeled with digoxigenin.

STORAGE CONDITIONS

Supplied reagents are stored at room temperature until their expiration date. After being reconstituted the probes will remain stable for two weeks at 4ºC in a DNAase-free environment.  The Anti-Digoxin can be stored at 4ºC for 1 month and the Proteinase K must be used immediately and cannot be stored.  Do not use these products after their expiration dates.

SAMPLES

Any paraffin block section in which Lysozyme RNA presence is to be studied. Sections of 4-6 micrometers in width are sufficient to conduct the study. Preferably, the cut should be recent (no more than thirty days old). Assay results are not affected by block age. Studies have been carried out in the manufacturers laboratories using 20 year old paraffin blocks with optimal results.

INTERPRETATION OF RESULTS

Samples in which Lysozyme expression is observed will show a brownish color in the cell cytoplasm, which will contrast over the blue-violet background given by hematoxylin staining. The pathologist will evaluate the results according to their experience, drawing conclusions from the staining of the sample in parallel with the staining observed in positive and negative controls.

ASSAY LIMITATIONS

The Kit Histosonda Lysozyme has been optimized to detect Lysozyme RNA expression in formalin-fixed, paraffin-embedded tissues. Its use is not recommended for other types of samples or preparation techniques.

The correct operation of this kit has been validated using the protocols indicated in the instructions manual. The use of other procedures or the modification of the recommended protocols may lead to erroneous results.

The results from this assay must be evaluated by the pathologist in combination with the rest of available patient clinical data.

In order to obtain optimal and reproducible results it is important to rigorously maintain the time and temperature conditions indicated in the procedure.