Applying Multiplex Real-time PCR to Evaluate E6/E7 Expression in Human Papillomavirus

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Applying Multiplex Real-time PCR to Evaluate E6/E7 Expression in Human Papillomavirus

14 Tháng 06, 2026

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Applying Multiplex Real-time PCR to Evaluate E6/E7 Expression in Human Papillomavirus

Van Hung Pham^1,2*, Viet Quoc Tran^1,2, Nhan Thien Vo^1,2,3, Huong Thien Pham^1,2,4
¹Vietnam Research and Development Institute of Clinical Microbiology, Ho Chi Minh city 70000, Vietnam. ²Nam khoa Biotek Co; ³International University, Vietnam National University, Ho Chi Minh city 70000, Vietnam; ⁴Pham Ngoc Thach University of Medicine, Ho Chi Minh city 70000, Vietnam

BACKGROUND

In HPV genotype detection testing, to definitively determine whether the detected HPV genotype isactually invading and proliferating within epithelial cells, it is essential to detect its E6 and/or E7 mRNA^[1,2,3]. Therefore, this is a test currently recommended to be included with HPV genotype detection testing, and we have developed and perfected this test to complement the HPV genotype detection test that we have been implementing

AIMS OF THE STUDY

This study aimed to determine the detection rate of E6 and E7 mRNA from cervical smear samples that had HPV genotype detection results and collected PAP smear samples that detected epithelial cell abnormalities, in order to assess the detection rate of cases where HPV genotypes actually invaded and proliferated in epithelial cells.

MATERIALS AND METHODS

Retrospective analysis was performed on 587 cervical cell samples at the Vietnam Research and Development Institute of Clinical Microbiology (VCM). Total RNA was extracted and processed with RNAse-free DNAse to ensure DNA removal. We used the reverse transcriptase multiplex real-time PCR (RT MLP-rPCR) targeting the detection of E6/E7 mRNA of 14 high-risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) and also 2 low-risk types (6, 11).

DISCUSSIONS AND CONCLUSION

Research results show that E6/E7 mRNA detection testing is essential when combined with HPV genotype detection testing to determine whether the pathogen has truly invaded the genital epithelial cells of patients. Therefore, in many countries, HPV E6/E7 mRNA detection testing is now recommended as a replacement for PAP smear testing, as PAP smear results are not truly specific for this purpose. However, because commercial E6/E7 mRNA testing products are still quite expensive, we hope that after this study and many future studies, this test will be more easily implemented due to its affordable cost.

KEY WORDS

HPV genotypes, E6/E7 mRNA; Multiplex real-time PCR (MPL-rPCR); Reverse transcriptase multiplex real-time PCR (RT MLP-rPCR).

REFERENCES

1. Janiszewska, J et al. HPV-driven oncogenesis—much more than the E6 and E7 oncoproteins. J Appl Genetics 66, 63–71 (2025). https://doi.org/10.1007/s13353-024-00883-y
2. Janicek MF, Averette HE (2001) Cervical cancer: prevention, diagnosis, and therapeutics. CA: Cancer J Clin 51(2):92–114. https://doi. org/10.3322/canjclin.51.2.92. quiz 115–18
3. Pankaj S, Rani J, Kumari P, Abhilashi K, Choudhary V, Kumari S, Shahi SK, Xess A, Singh RK, Kumar S, Kumar A, Biswas NR, Jee B. Efficacy of HPV E6/E7 mRNA assay, HPV DNA test and cytology in detection of high grade cervical lesions and invasive cancer at a tertiary care center in India. Malawi Med J. 2024 Jul 30;36(2):120-127. doi: 10.4314/mmj.v36i2.9. PMID: 40191563; PMCID:PMC11970201.

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Applying Multiplex Real-time PCR to Evaluate E6/E7 Expression in Human Papillomavirus

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