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Updated: Mar 28, 2026
RNAscope for In situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma
Published on: March 11, 2014
Optimization of RNAscope for HPV mRNA detection in liquid-based cervical cytology samples
Tianqi Liu1, Xianrong Zhou1, Xiang Tao1
1Department of Pathology, Obstetrics & Gynecology Hospital of Fudan University, Shanghai Key Lab of Reproduction and Development, Shanghai Key Lab of Female Reproductive Endocrine Related Diseases, Shanghai, China.
Purpose:
To establish an RNAscope-based joint detection scheme compatible with liquid-based cytology (LBC) slides and staining systems, enabling simultaneous observation of cytological phenotype and HPV E6/E7 mRNA expression in cervical cancer and precancerous lesions.
Methods:
From March 2023 to February 2024, 115 LBC samples were collected at the Affiliated Obstetrics and Gynecology Hospital of Fudan University. Histopathological diagnosis included 30 LSIL, 33 HSIL, 29 HPV-related SCC, 8 HPV-related adenocarcinoma and 15 negative for cervical intraepithelial lesions cases. RNAscope technology, originally applied to paraffin samples, was adapted for LBC by modifying slide type selection, extending target retrieval, and enhancing permeabilization. The improved RNAscope in situ hybridization was then employed to detect high-risk HPV E6/E7 mRNA. Positive rates, diagnostic concordance with histopathology and, expression pattern were statistically analyzed.
Results:
RNAscope combined with LBC showed high positive rates, 93.33% for LSIL, 96.97% for HSIL, 100.00% for SCC, and 87.50% for HPV-related adenocarcinoma. Compared with HPV PCR testing, the diagnostic concordance rate reached 95.65% (P<0.001). Figures illustrate RNAscope positive expression patterns in LSIL, HSIL, SCC and, HPV-related adenocarcinoma.
Conclusion:
The adaptation of RNAscope technology, originally designed for paraffin samples, enables accurate diagnosis in cervical LBC, enhances consistency with histopathology, and provides a novel diagnostic approach for cervical lesions.
