CENBIMO S.L

Ghrelin is an endogenous ligand for the secretagogue receptor of the pituitary growth hormone.  Neuroendocrine tumors producing Ghrelin have been described as clinically developing with acromegaly.

The notable functions of Ghrelin, amongst others are its orexigenic action (stimulation of the ingestion of alimentation) and its stimulus of hydrochloric acid production by the stomach parietal cells.

Many studies have related some obese patients with a seric increase of Ghrelin.  Some publications have demonstrated that Ghrelin plays a part in inhibiting the production of leptin.

Ghrelin is produced by neuroendocrine cells that are preferably situated in the hydrochloric acid producing glands of the gastric body.

Some authors have also described cells that produce Ghrelin in other anatomic localizations although an exhaustive “in situ” hybridization map has not yet been made.

Cenbimo has found cases of hyperplasia and tumors of gastric neuroendocrine cells that produce Ghrelin (data not yet published).

Histosonda GHRL consists of a fragment of single stranded DNA, 241 nucleotides in length targeted against Ghrelin RNA.

The use of this probe is the detection of cells producing this hormone and their tumors.

INTENDED USE

For use in In Vitro Diagnosis.

The Kit Histosonda GHRL is useful for the detection of cells producing this hormone and their tumors.

WARNINGS AND PRECAUTIONS

The Kit Histosonda GHRL 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 GHRL; 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 GHRL consists of a single stranded DNA fragment with a length of 241 nucleotides targeted against Ghrelin 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 Ghrelin 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 Ghrelin 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 GHRL has been optimized to detect Ghrelin 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.

DOWNLOAD DATASHEET