CENBIMO S.L

INTRODUCTION

18s RNA is a constitutive of the small subunit of human ribosomes.  Its expression is continuous and forms a part of the large complexes of ribonucleoproteins dedicated to translation in these organelles.  Histosonda 18s RNA consists of a hybridization probe formed by a single stranded DNA fragment 354 nucleotides in length which is complementary to the transcribed RNA of the gene 18s RNA.

The use of this probe is to assess suitability of tissues for hybridization and to assess the quality of RNA that remains in the tissues.  The use of this probe is to detect cytoplasmic expression in all types of cells.

INTENDED USE

For use in In Vitro Diagnosis.

The Kit Histosonda 18s RNA is useful to asses the suitability of tissues for hybridization and to assess the quality of RNA that remains in the tissues.

WARNINGS AND PRECAUTIONS

The Kit Histosonda 18s RNA 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 18s RNA; 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 18s RNA consists of a single stranded DNA fragment with a length of 354 nucleotides targeted against 18s RNA mRNA.  The DNA of these probes 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 18s 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 18s RNA 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 which should always be positive.  In case of negativity, the sample should be considered as unsuitable for in situ hybridization.

ASSAY LIMITATIONS

The Kit Histosonda 18s RNA has been optimized to detect 18s 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.