Background Molecular analysis for common somatic mutations in thyroid cancer can improve diagnostic accuracy of fine-needle aspiration cytology (FNAC) in the nondiagnostic or indeterminate category of thyroid nodules. analysed for rearrangements and there was no abnormality. Conclusion Molecular analysis for and mutations was feasible in residual LBC materials and might be useful for diagnosis of indeterminate thyroid nodules. mutations were found in cases classified cytologically as malignant or suspicious for malignancy . However, the usefulness of testing was limited by the low rate of point mutations and rearrangements of using residual SurePath samples of cytological nondiagnostic or indeterminate thyroid nodules such as AUS/FLUS or FN/SFN categories. METHODS Patients and samples This prospective study enrolled 53 thyroid nodules from 53 patients, which were diagnosed nondiagnostic or indeterminate thyroid nodules after thyroid FNAC from July 2014 to September 2014 in Asan Medical Center. The thyroid nodules were classified as 121268-17-5 non-diagnostic ((codon 600), (codon 61), (codon 12/13), and (codon 61) point mutations were detected by real-time polymerase chain reaction (RT-PCR) and high-resolution melting (HRM), using the LightCycler 480 High Resolution Melting Grasp chemistry (Roche, Mannheim, Germany) on a LightCycler 480 (Roche) 121268-17-5 as previously reported [11,13,15]. rearrangements were also evaluated after cDNA synthesis from RNAs. PCRs were performed as previously reported . Detection of point mutations and rearrangements by pyrosequencing (codon 600), (codon 61), (codon 12/13), and (codon 61) point mutations were detected by pyrosequencing on a PyroMark Q24 (Qiagen) according to the manufacturer’s instructions. Pyrosequencing was applied if the samples were considered as mutation positive or showed ambiguous results by HRM peak analysis. When the samples were decided wild-type by HRM peak analysis, some samples of them were analyzed by pyrosequencing as previously reported . Statistical analysis Statistical analysis was conducted by using SPSS version 21.0 (IBM Co., Armonk, NY, USA). Graphs were produced using Prism version 5.01 (GraphPad Software Inc., La Jolla, CA, USA). Continuous variables such as age and primary nodule size were presented as meanstandard deviation. RESULTS Evaluation for or point 121268-17-5 mutations All DNAs from 53 residual LBC samples could be analysed for point mutations by RT-PCR or pyrosequencing. Point mutations were detected in 10 of 53 samples (19%) (Table 1, Fig. 1). There were no or point mutations in 16 nondiagnostic thyroid nodules. In 17 AUS thyroid nodules, seven samples (41%) had point mutations including ((((point mutation was detected in sample A2, A6, A9, and A16 by pyrosequencing (mutation percentage 10%, 12%, 19%, and 14%, respectively). point mutations were detected in sample A5 (point mutation in sample A15 (mutation percentage 16%). There were no mutations in AUS thyroid nodules. In FLUS thyroid nodules, point mutations were detected in sample F2 (mutations in FLUS thyroid nodules. Evaluation for rearrangements Only one RNA sample from a FLUS thyroid nodule could be analysed for rearrangements (Fig. 2). There were no rearrangements in this sample. The other RNA samples were not amplified by PCR. Fig. 2 Only one RNA sample from a follicular lesion of undetermined significance (FLUS) thyroid nodule could be analysed for rearrangements. AUS, atypia of undetermined significance DISCUSSION In this study, molecular analysis for and point mutations was feasible using residual supernatant of LBC SurePath samples after FNA. We found or 121268-17-5 point mutations in 10 of 37 AUS/FLUS nodules (27%). However, we did not find point mutations in nondiagnostic thyroid nodules, and RNAs from residual SurePath material were not applicable for molecular analysis of gene rearrangements. FNA is the procedure of Mouse monoclonal antibody to PEG10. This is a paternally expressed imprinted gene that encodes transcripts containing twooverlapping open reading frames (ORFs), RF1 and RF1/RF2, as well as retroviral-like slippageand pseudoknot elements, which can induce a -1 nucleotide frame-shift. ORF1 encodes ashorter isoform with a CCHC-type zinc finger motif containing a sequence characteristic of gagproteins of most retroviruses and some retrotransposons. The longer isoform is the result of -1translational frame-shifting leading to translation of a gag/pol-like protein combining RF1 andRF2. It contains the active-site consensus sequence of the protease domain of pol proteins.Additional isoforms resulting from alternatively spliced transcript variants, as well as from use ofupstream non-AUG (CUG) start codon, have been reported for this gene. Increased expressionof this gene is associated with hepatocellular carcinomas. [provided by RefSeq, May 2010] choice in the evaluation of thyroid nodules . Once the needle is usually withdrawn from the lesion, the material is usually extruded onto glass slides for a conventional smear which is usually fixed with 95% ethyl alcohol for the Papanicolau stain. Otherwise, the smear may be air-dried and then stained with May-Grunwald Giemsa. The thin-layer or LBC technique has been used popularly as an alternative technique for conventional smear due to cost-effectiveness, time-sparing, and simple application with a semi-automated device [10,12]. It is originally.