Genetic tests must be requested by doctors and healthcare professionals working in affiliated hospitals.
Before submitting a request, the doctor shall:
ensure that the test is appropriate for the patient on the basis of the pathological findings and his or her medical history
inform the patient about the meaning, the limits and consequences of the test
obtain the patient's informed consent for the test execution
The test is performed on DNA extracted from peripheral blood samples.
Upon arrival of such biological samples in our laboratories, a quality check is performed to verify if the samples care eligible for the analysis.
The mutation search is then performed using the "resequencing" technique that compares the sequence obtained from the sample with a standard sequence (the normal or "wild type" sequence).
Recent technological developments have made such resequencing technique extremely fast, accurate and reliable.
The protocol in use in our laboratory for searching mutations of interest involves the use of an amplification reaction (PCR) isothermal for all coding regions. The amplification conditions are optimized and standardized in order to obtain consistent results.
Amplified products are then directly sequenced and analyzed for mutation search through a dedicated software.
The test is then completed with the MPLA ("Multiplex Ligation-dependent Probe Amplification") technique to ensure that there are no gene rearrangements if sequencing analysis was negative.