时间分辨荧光法检测宫颈脱落细胞p16^(INK4a)蛋白用于宫颈肿瘤筛查

Application of detection of p16^(INK4a) protein in cervical exfoliated cells by time-resolved fluoroimmunoassay to screening of cervical neoplasm

目的应用时间分辨荧光法(TRFIA)检测宫颈脱落细胞p16^(INK4a)蛋白,将其应用于宫颈肿瘤筛查。方法搜集2015年1月-2016年5在荆门市第一人民医院病理科进行宫颈组织学活检并进行过TCT和/或DNA倍体分析检测标本,宫颈脱落细胞离心甩片进行TCT检测,Feulgen染色行DNA倍体分析,TRFIA方法检测p16^(INK4a)蛋白。结果 694例标本中组织病理学诊断宫颈炎性病变501例,宫颈肿瘤性病变193例,包括低级别上皮内病变(LSIL)83例、高级别上皮内病变89例(HSIL)、鳞状细胞癌(SCC)21例。TCT检测筛查宫颈肿瘤敏感性为76.68%,特异性为87.03%;DNA倍体分析检测敏感性84.97%(优于TCT,χ2=4.280,P=0.039),特异性73.45%(低于TCT,χ~2=29.105,P<0.001)。TRFIA检测p16^(INK4a)筛查宫颈肿瘤敏感性为86.01%,与DNA倍体分析相似(χ~2=0.084,0=0.773),优于TCT检测(χ~2=5.532,P=0.019),且特异性达92.42%,亦优于TCT检测(χ~2=7.889,P=0.005)。结论时间分辨法检测宫颈脱落细胞p16^(INK4a)蛋白用于宫颈肿瘤筛查,敏感性及特异性均较好。如果试剂盒商品化,还具有操作简便、成本低廉、能用仪器自动化检测并给出客观判读结果的优点。

Objective To apply detection of p16INK4a protein in cervical exfoliated cells by time-resolved fluoroimmunoassay （TRFIA） to screening cervical neoplasm. Methods We collected the specimens with cervical biopsy, thinprep cytologic test （TCT） and/or DNA ploidy analysis in Department of Pathology, the First People’s Hospital of Jingmen City from January, 2015 to May, 2016. The cervical exfoliated cells were centrifuged and dropped on the films for TCT. DNA ploidy analysis was performed after Feulgen staining, and TRFIA was used to detect p16INK4a protein. Results Among 694 specimens diagnosed by histopathology, there were 501 cases with cervicitis and 193 with cervical neoplasia （including 83 cases with low grade squamous intraepithelial lesion （LSIL）, 89 with high grade squamous intraepithelial lesion （HSIL） and 21 with squamous cell carcinoma （SCC）. The sensitivity and specificity of TCT for screening of cervical neoplasm were 76.68% and 87.03%. The sensitivity（84.97%） of DNA ploidy was superior to that of TCT（χ^2=4.280,P=0.039） , while the specificity（73.45%） was inferior（χ^2=29.105,P〈0.0001）. The sensitivity of p16INK4a detection via TRFIA was 86.01%, similar to that of DNA ploidy analysis （χ^2=0.084, P=0.773） but superior to that of TCT （χ^2=5.532, P=0.019）, and the specificity was 92.42% , superior to that of TCT （χ^2=7.889, P=0.005）. Conclusions Detection of p16INK4a protein in cervical exfoliated cells by TRFIA displays a high sensitivity and specificity for screening cervical neoplasm . In addition, it has the advantages of simple operation, low cost, automatic detection and providing objective judgment results if the kit reaches commercialization level.