Multiplex PCR approach to simultaneously identify several mutations in fine needle cytology thyroid samples
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Emilia Vuttariello1, Marco Borra2, Elvira Mauriello2, Celeste Calise3, Barbara D’Andrea2,3, Anna Capiluongo1, Franco Fulciniti4, Anna Cipolletta5, Mario Monaco1, Luciano Pezzullo6 and Gennaro Chiappetta1
1Functional Genomics Unit, Istituto Nazionale Tumori, IRCCS, Fondazione G. Pascale, Naples, Italy
2Molecular Biology and Bioinformatics Unit, Stazione Zoologica “A.Dorhn”, Naples, Italy
3CMO, Naples, Italy
4Clinical Cytopathology Service, Institute of Pathology, Locarno, Switzerland
5SC Anatomia Patologica e Citopatologia, Istituto Nazionale Tumori, IRCCS, Fondazione G. Pascale, Naples, Italy
6Thyroid Surgery Unit, Istituto Nazionale Tumori, IRCCS, Fondazione G. Pascale, Naples, Italy
Emilia Vuttariello, email: firstname.lastname@example.org
Gennaro Chiappetta, email: email@example.com
Keywords: multiplex PCR, thyroid, fine needle cytology, genetic testing, Sanger sequencing
Received: November 24, 2016 Accepted: April 19, 2017 Published: May 07, 2017
The most frequent initial manifestation of thyroid cancer is the appearance of a nodule. More than 20% of the general population has a palpable thyroid nodule and the percentage rises to 70% based on ultrasound identification. In 95% of cases the nodule is simply a hyperplastic or benign lesion. The most reliable diagnostic test for thyroid nodules is fine needle aspiration (FNA), but cytological discrimination between malignant and benign follicular neoplasms remains difficult. Cytological analysis is now, almost routinely, being combined with molecular genetics to enable the pathologist to make a more objective diagnosis. In this study, we performed the molecular analysis using a new simplified procedure that involves a panel of BRAF, RAS, RET and RET/PTC gene mutations in easily obtainable FNA samples, in the attempt to improve the efficacy of the FNA diagnosis of thyroid nodules and thus patient management. In this new procedure, PCR and sequencing analysis are used to detect point mutations, and, in parallel, RT-PCR is used to detect the chimeric RET/PTC1 and RET/PTC3 transcripts in RNA extracted from FNA.
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