Histosonda®

INTRODUCTION

Calcitonin is a peptide of 32 amino acids and 3.4 kDa produced by the thyroid parafollicular cells or C cells.

Calcitonin is coded by a gene named calcitonin gene-related peptide (CGRP) that also includes the code for a peptide related to calcitonin but with a completely different physiological function.  The production of one or the other peptide is due to a splicing alternative.

The principle physiological function of this hormone is to produce a decrease in the blood concentration of calcium and phosphates producing an incorporation of these ions into the bones.

The tumoral degeneration of C cells produces a Thyroid Medullary Carcinoma also known as Thyroid Parafollicular Carcinoma.

Approximately 70% of Thyroid Medullary Carcinomas are sporadic and approximately 30% are familial and inheritable within the context of MEN 2A.  MEN 2A patients can also demonstrate C cell hyperplasia before developing a tumor.

INTENDED USE

For use in In Vitro Diagnosis.

The Kit Histosonda Calcitonin is useful for the diagnosis of Thyroid Medullary Carcinomas and C cell Hyperplasia.

WARNINGS AND PRECAUTIONS

The Kit Histosonda Calctionin 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 Calcitonin; 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 Calcitonin consists of two segments of single-stranded DNA complementary to expressed RNA with lengths of 166 and 142 nucleotides. 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 Calcitonin 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 manufacturer's laboratories using 20 year old paraffin blocks with optimal results.

INTERPRETATION OF RESULTS

Samples in which Calcitonin 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 Calcitonin has been optimized to detect 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.

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