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Article Open Access

Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia

  • Authors:
    • Stéphanie Calfa
    • Ana Julia Aguiar De Freitas
    • Welinton Hirai
    • Júlio César Possati‑Resende
    • Rhafaela Lima Causin
    • Ricardo Dos Reis
    • Rui Manuel Reis
    • Márcia Maria Chiquitelli Marques
  • View Affiliations / Copyright

    Affiliations: Molecular Oncology Research Center, Teaching and Research Institute, Barretos Cancer Hospital, Barretos, Sao Paulo 14784‑400, Brazil, Nucleus of Epidemiology and Biostatistics, Barretos Cancer Hospital, Barretos, Sao Paulo 14784‑400, Brazil, Department of Prevention, Barretos Cancer Hospital, Barretos, Sao Paulo 14784‑400, Brazil, Department of Gynecologic Oncology, Barretos Cancer Hospital, Barretos, Sao Paulo 14784‑400, Brazil
    Copyright: © Calfa et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 53
    |
    Published online on: November 26, 2025
       https://doi.org/10.3892/ol.2025.15406
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Abstract

Liquid biopsy offers a minimally invasive approach for cancer diagnosis by detecting circulating biomarkers, such as microRNAs (miRNAs), in different types of specimens. The present study aimed to identify the most effective specimen for miRNA detection in cervical cancer precursor lesions by comparing plasma, urine and liquid‑based cytology (LBC). A total of 798 miRNAs were analyzed using NanoString technology in 24 women, equally divided between those with high‑grade cervical intraepithelial neoplasia (2/3) and controls with negative Pap and human papillomavirus tests. Comparative analyses revealed that LBC exhibited the highest detection efficiency, with only 1.75% of miRNAs demonstrating low counts, compared with 20.68% in plasma and 15.79% in urine. Additionally, LBC exhibited 21.3% of miRNAs above the 9th decile of expression, compared with 5.89% in plasma and 2.88% in urine. LBC revealed the most consistent detection performance across samples, establishing its potential as the most effective specimen for detecting cervical cancer precursor lesions biomarkers. Plasma also showed promise as a detection medium, whilst urine exhibited higher variability and lower detection consistency. In conclusion, LBC demonstrated the highest efficacy for miRNA detection among the specimen types tested. Plasma remains a viable alternative, whilst urine presents challenges due to its inherent variability. These results underscore the importance of specimen selection for optimizing diagnostic sensitivity in cervical cancer precursor lesions screening.
View Figures

Figure 1

Study overview. Figure created using
BioRender (https://www.biorender.com/). CIN,
cervical intraepithelial neoplasia/lesion; LEEP, Loop
Electrosurgical Excision Procedure; HPV, human papillomavirus; LBC,
liquid-based cytology; miRNA, microRNA.

Figure 2

Raw data count comparison chart of
798 miRNAs between the three specimens. (A) Plasma. (B) Urine. (C)
LBC. The y-axes present the proportions of counts (<10) of each
miRNA in the raw data, and the x-axes present the 798 miRNAs. The
red lines indicate 50% of the samples. miRNA, microRNA; LBC,
liquid-based cytology.

Figure 3

Raw data count comparison chart of
798 between the case and control groups for each specimen. (A)
Plasma. (B) Urine. (C) LBC. The y-axes present the proportions of
counts (<10) of each miRNA in the raw data, and the x-axes
present the 798 miRNAs. The red lines indicate 50% of the samples.
LBC, liquid-based cytology; miRNA, microRNA.

Figure 4

Based on normalized data, density
curves show the mean count values for the 798 miRNAs. (A) LBC vs.
plasma. (B) LBC vs. urine. (C) Plasma vs. urine. The y-axes present
the density values, whilst the x-axes present the sample mean
values of the detected miRNAs. The red lines indicate the 9th
decile. miRNA, microRNA; LBC, liquid-based cytology.

Figure 5

Comparison of LBC, plasma and urine
for the detection of miRNA biomarkers in cervical cancer precursor
lesions. The gradient arrow indicates the classification of
specimens from more to less effective using NanoString technology.
Figure created using BioRender (https://www.biorender.com/). LBC, liquid-based
cytology; miRNA, microRNA.
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Copy and paste a formatted citation
Spandidos Publications style
Calfa S, De Freitas AA, Hirai W, Possati‑Resende J, Causin RL, Dos Reis R, Reis RM and Marques MC: Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia. Oncol Lett 31: 53, 2026.
APA
Calfa, S., De Freitas, A.A., Hirai, W., Possati‑Resende, J., Causin, R.L., Dos Reis, R. ... Marques, M.C. (2026). Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia. Oncology Letters, 31, 53. https://doi.org/10.3892/ol.2025.15406
MLA
Calfa, S., De Freitas, A. A., Hirai, W., Possati‑Resende, J., Causin, R. L., Dos Reis, R., Reis, R. M., Marques, M. C."Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia". Oncology Letters 31.2 (2026): 53.
Chicago
Calfa, S., De Freitas, A. A., Hirai, W., Possati‑Resende, J., Causin, R. L., Dos Reis, R., Reis, R. M., Marques, M. C."Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia". Oncology Letters 31, no. 2 (2026): 53. https://doi.org/10.3892/ol.2025.15406
Copy and paste a formatted citation
x
Spandidos Publications style
Calfa S, De Freitas AA, Hirai W, Possati‑Resende J, Causin RL, Dos Reis R, Reis RM and Marques MC: Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia. Oncol Lett 31: 53, 2026.
APA
Calfa, S., De Freitas, A.A., Hirai, W., Possati‑Resende, J., Causin, R.L., Dos Reis, R. ... Marques, M.C. (2026). Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia. Oncology Letters, 31, 53. https://doi.org/10.3892/ol.2025.15406
MLA
Calfa, S., De Freitas, A. A., Hirai, W., Possati‑Resende, J., Causin, R. L., Dos Reis, R., Reis, R. M., Marques, M. C."Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia". Oncology Letters 31.2 (2026): 53.
Chicago
Calfa, S., De Freitas, A. A., Hirai, W., Possati‑Resende, J., Causin, R. L., Dos Reis, R., Reis, R. M., Marques, M. C."Comparative evaluation of microRNA detection in plasma, urine and liquid‑based cytology for high‑grade cervical intraepithelial neoplasia". Oncology Letters 31, no. 2 (2026): 53. https://doi.org/10.3892/ol.2025.15406
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