<p>A Sensitive and Simplified Classifier of Cervical Lesions Based on a Methylation-Specific PCR Assay: A Chinese Cohort Study</p>

Abstract

These authors contributed equally to this workObjective: The aim of this study is to assess the diagnostic and screening performance of a standardized methylation-specific real-time PCR assay targeting SOX1 and PAX1 genes for cervical cancer in a Chinese cohort. Methods: Genomic DNA was extracted from cervical exfoliated cells and converted by sodium bisulfite and then analyzed by qMSP assay. Ct values were collected for PAX1 and SOX1 as target genes and β-actin as an endogenous reference gene. The samples included 295 cervicitis, 111 LSIL (low-grade squamous intraepithelial lesion), 51 HSIL (high-grade squamous intraepithelial lesion) and 30 cervical cancer. Results: The Ct values decreased with the progression of cervical cancer from cervicitis, through LSIL and HSIL to cancer. The difference in Ct values between cytological grades was highly significant (p≤0.01) between grades either for PAX1 or for SOX1 except the difference between cervicitis and LSIL of SOX1. With the Ct cut-off values of PAX1 gene and SOX1 gene 38.6 and 38 and with the PAX1/SOX1 in combination, the positive rate of methylation in invasive cancer tissues was 100%, in contrast to 11.5% (95% CI: 8.67%-14.33%) in cervicitis tissues, 45.1% (95% CI: 40.68%-49.52%) in LSIL tissues, and 68.5% (95% CI: 64.37%-72.63%) in HSIL tissues. The specificity and sensitivity of differentiating tumors from cervicitis were 0.957 (95% CI: 0.939-0.975) and 1.00, respectively. The specificity and sensitivity of differentiation between cervicitis+LSIL and HSIL+cervical cancer were 0.881 (95% CI: 0.852-0.91) and 0.748 (95% CI: 0.709-0.787), respectively. Conclusion: PAX1/SOX1 methylation could be translated into clinical practice for cervical neoplasia detection.

Analysis of the diagnostic performance of PAX1/SOX1 gene methylation in cervical precancerous lesions and its role in triage diagnosis

“…In the ASCUS group, nine cases were identified as methylation-positive for CIN2+, with no methylationnegative CIN2+ cases. Overall, 37 cases were methylation-positive, resulting in a colposcopy referral rate of 22.29% (37 out of 166) 16. The current study confirmed these trends, with methylation positivity rates increasing with lesion severity: 17.1% in control group, 22.5% in CIN1 group, 100% in CIN2 group, 90% in CIN3 group, and 100% in CC group.…”

“…Our prior research on PAX1/SOX1 gene methylation in cervical cells from 487 patients showed that methylation rates was 100% in cancer group, 11.5% in cervical inflammation group, 45.1% in LSIL group, and 68.5% in HSIL group 16 . The current study confirmed these trends, with methylation positivity rates increasing with lesion severity: 17.1% in control group, 22.5% in CIN1 group, 100% in CIN2 group, 90% in CIN3 group, and 100% in CC group.…”

“…However, there is limited research on the application of the SOX1 and PAX1 gene methylation panel for cervical lesion diagnosis. While our research team has previously established the cut‐off values for these genes, 16 further investigation into their diagnostic performance across various cervical lesion stages is needed. Additionally, the role of triage in patients with ASCUS remains unclear.…”

A dual-gene panel of two fragments of methylated IRF4 and one of ZEB2 in plasma cell-free DNA for gastric cancer detection

“…The methylation index, evaluated with MSP, has been the most frequently used method to assess the methylation level. There are several methods for calculating methylation levels, including MI = ΔCt [ 50 ], MI = 2 −ΔCt [ 51 ], MI = 10,000 × 2 ΔCt where ΔCt = Ct target gene - Ct internal reference [ 52 ], MI = 2 −ΔΔCt where ΔΔCt = ΔCt sample - ΔCt calibrator [ 51 ], MI = Ct target gene /Ct internal reference [ 52 ], and MI = Ct [ 26 , 41 , 53 ]. Regarding the use of Ct as an MI index, the lower the Ct value is, the higher the methylation level is, and the endogenous reference such as ACTB used in this study should be valid, indicating that the sample treatment was effective.…”

The Role of the von Willebrand Factor Collagen-Binding Assay (VWF:CB) in the Diagnosis and Treatment of von Willebrand Disease (VWD) and Way Beyond: A Comprehensive 36-Year History

“…20 Ezigbo et al 256 included the VWF:CB to help characterize VWD in the Nigerian population. Zhang et al 257 included the VWF:CB in their study of VWF level and activity in healthy Chinese. Heilmann et al 258 included the VWF:CB in their study of AVWS in 74 patients on long-term support with the VAD HeartMate II.…”