AI Article Synopsis
- The study investigates the effectiveness of droplet digital PCR (ddPCR) compared to Sanger sequencing in detecting the exon 15 p.V600E mutation in thyroid fine-needle aspiration (FNA) samples.
- In a sample of 310 thyroid nodules, ddPCR identified the mutation in 30.32% of cases, while Sanger sequencing detected it in only 12.90%, highlighting ddPCR's superior sensitivity.
- The authors recommend using ddPCR alongside Sanger sequencing for more accurate molecular testing in diagnosing papillary thyroid carcinoma from FNAB samples.
Article Abstract
Background: exon 15 p.V600E ( V600E) mutation has been established as an important molecular marker for papillary thyroid carcinoma diagnosis by ultrasound-guided fine-needle aspiration biopsy (FNAB). Sanger sequencing is the gold standard for detecting V600E mutations but fails to identify low-frequency mutations. However, droplet digital PCR (ddPCR) is a popular new method for detecting low-frequency mutations. Here, we compare the efficiency of droplet digital PCR (ddPCR) and Sanger sequencing for detection of the V600E mutation in thyroid fine-needle aspiration (FNA) samples.
Methods: Thyroid fine-needle aspiration samples from 278 patients with 310 thyroid nodules were collected. Sanger sequencing and ddPCR were conducted to detect the V600E mutation.
Results: The V600E mutation was found in 94 nodules (30.32%) by ddPCR and 40 nodules (12.90%) by Sanger sequencing in 310 FNA samples. A total of 119 nodules were confirmed PTC by postsurgical pathology. Among which the mutation was found in 80 (67.23%) nodules by ddPCR and 31 (26.05%) by Sanger sequencing. All nodules carrying the mutation detected by Sanger sequencing (SS+) were verified by ddPCR (ddPCR+). Also, all nodules with no mutation detected by ddPCR were interpreted as wild-type by Sanger sequencing (SS-). In addition. Almost all SS+/ddPCR + nodules (95.00%; 38/40) and SS-/ddPCR + nodules (100.00%; 54/54) displayed a mutation rate of >5% and <15%, respectively, indicating easy misdetection by Sanger sequencing when the mutation rate is between 5 and 15%.
Conclusion: ddPCR has higher sensitivity than Sanger sequencing and we propose ddPCR as a supplement to Sanger sequencing in molecular testing of using FNAB samples.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8983273 | PMC |
| http://dx.doi.org/10.1155/2022/6243696 | DOI Listing |
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