Therefore, detection of Cyfra 21-1 in lymph nodes may predict the metastasis of cancer cells to lymph nodes. Recent evidence suggests that the level of Cyfra 21-1 is associated with distant metastasis and tumor progression of thyroid cancer ( Giovanella et al., 2008 Malapure et al., 2020). Elevated serum Cyfra 21-1 is an indicator of poor prognosis for many malignant neoplasms, including lung cancer, bladder cancer, and cervical cancer ( Ma et al., 2015 Chen et al., 2018 Jin et al., 2019 Mei et al., 2019). Cyfra 21-1 is the proteolytic portion of CK-19. Many studies have shown that CK-19 is highly expressed in differentiated thyroid carcinoma but is expressed at low or even no levels in benign thyroid tumors ( Arcolia et al., 2017 Viana et al., 2020 Menz et al., 2021). Cytokeratin 19 (CK-19) is a low molecular weight cytokeratin that exists in a variety of tumors of epithelial and normal epithelial origin ( Chen et al., 2018). Metastasis can be determined by detecting markers of cancer cells in lymph nodes. Thyroid tumor cells are destroyed during the metastasis cascade, releasing many tissue structural components into body fluids ( Li et al., 2006). Tumor metastasis is a continuous and dynamic process involving the separation of tumor cells from the site of origin. Therefore, rapid, accurate, and quantitative intraoperative detection of lymph node metastasis is of great importance to improve the quality of thyroid cancer surgery and the prognosis of patients with this cancer. If a rapid detection method can be developed to accurately determine intraoperative lymph node metastasis, the need for further lymph node dissection could be determined during surgery, which may reduce the opportunity for tumor metastasis and multiple operations. The results of frozen section and postoperative pathological examinations are dependent more on the subjective judgment of the pathologist than on objective findings ( Roberts et al., 2003). In addition, numerous studies have shown that the false negative rate of intraoperative histopathological examination is 5%–52% ( Celebioglu et al., 2006 Zavagno et al., 2008 Horvath et al., 2009 Hoen et al., 2016). Obtaining results from frozen sections takes approximately 60 min during surgery, whereas obtaining postoperative pathological results typically takes 5–10 days.
Detection of lymph node metastasis includes the use of rapid intraoperative frozen sections and postoperative histopathological examination. Although many of these cases had a low degree of malignancy, approximately 40%–60% of patients with thyroid cancer experience early cervical lymph node metastasis ( Smith et al., 2012 Tang et al., 2018). In 2020, there were 586,202 newly diagnosed thyroid cancer cases worldwide, making it one of the most common endocrine tumors ( Sung et al., 2021). Together, these results indicated that although further testing using a larger sample size will be required, this immunochromatographic strip test may be useful for rapid intraoperative detection of thyroid cancer metastasis to lymph nodes. The sensitivity of the strip remained stable when stored at room temperature for 6 months. In an assessment of 20 lymph node specimens that had been determined by postoperative histopathology to be positive for lymph node metastasis and 20 specimens that were negative, the sensitivity and specificity of the strip were 100% and 95%, respectively. The ability of the strip to differentiate positive from negative metastasis in 40 lymph node specimens was 100% concordant with that of immunohistochemical staining for Cyfra 21-1. The strip did not detect other protein markers of epithelial cells at a concentration of 500 ng mL −1, including cytokeratin 8, cytokeratin 18, epithelial membrane antigen, and epidermal surface antigen. The standard (calibration) curve equation was Y = 0.003708 × X + 0.1101, and the detection limit was 0.55–1.14 ng mL −1. In this study, an immunochromatographic strip test based on colloidal gold nanoparticles was developed to semi-quantitatively detect the levels of Cyfra 21-1 in lymph nodes within 15 min. Elevated levels of Cyfra 21-1, the proteolytic portion of cytokeratin, are associated with the metastasis and progression of thyroid cancer and are an effective biomarker for the prognosis and diagnosis of thyroid cancer. Currently, detection of lymph node metastasis of thyroid cancer during surgery is limited and time-consuming. Thyroid cancer is the most common endocrine tumor, and the rate of early lymph node metastasis may be as high as 60%.
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