Recently, Professor Lina Zhang and colleagues from Changzhou Maternal and Child Health Hospital published a retrospective analysis titled "Correlation of different HPV genotype viral loads and cervical lesions: A retrospective analysis of 1585 cases" in the Journal of Cancer Cytopathol. This study aimed to explore the differences in human papillomavirus (HPV) viral loads across various cervical lesions and identify the optimal cutoff value for high‐grade squamous intraepithelial lesions (HSILs), and to explore an effective detection method to triage HPV-positive patients and reduce unnecessary examinations and treatments.

Conclusions

High‐risk HPV viral load is associated with cervical lesion classification. HPV16 viral load can effectively differentiate high‐grade squamous intraepithelial lesions (HSIL) from low‐grade squamous intraepithelial lesions (LSIL) with good sensitivity and specificity.

Methods

The pathological results of patients with negative low‐grade squamous intraepithelial lesions (LSILs) and high‐grade squamous intraepithelial lesions (HSILs) between January 1, 2023, and March 1, 2024, were retrospectively retrieved through the hospital’s pathological system. A total of 1585 patients were included in the study, comprising 666 with negative test results, 708 with LSILs, and 211 with HSILs.

The BioPerfectus multiplex real‐time (BMRT) detection method (Jiangsu BioPerfectus Technologies Co., Ltd.) was used to detect the L1 gene of 21 HPV genotypes, including 14 high‐risk HPV types (HPV16, HPV18, HPV31, HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV56, HPV58, HPV59, HPV66, and HPV68), four medium‐risk HPV types (HPV26, HPV53, HPV82, and HPV73), and three low‐risk HPV types (HPV6, HPV11, and HPV81). The human TOP3 single‐copy gene was used to standardize the number of sampled cells for quantifying each HPV type (Shanghai Hongshi PCR Instrument).

Results

01

As shown in Table 1, no significant difference was observed between the ages of the three groups of women. The negative group had the highest proportion of women with single infections (70%), whereas the LSIL group had the highest proportion of women with multiple infections (38.1%). Multiple HrHPV infections did not significantly increase the risk of developing HSIL compared with the corresponding single‐genotype infection. The reduced prevalence of multiple infections among women with HSIL supports the concept of intergenotypic competition among HrHPV genotypes in cervical squamous lesions. Furthermore, a significant difference was observed in the average viral load of the 21 HPV genotypes among the three groups.

02

Table 2 provides the prevalence of specific HPV genotypes. Regardless of the calculation method used, the top three HPV genotypes in terms of negative prevalence were HPV52, HPV16, and HPV58. For HSIL prevalence, the leading genotypes were HPV16, HPV58, and HPV52. Although the prevalence of HPV genotypes calculated using the three methods differed slightly for LSIL, the top three genotypes remained unchanged. The prevalence of HPV16 in the HSIL group was significantly higher than that in the negative and LSIL groups. Similarly, the prevalence of HPV58 in the HSIL group was slightly higher than that in the negative and LSIL groups. Conversely, HPV52 was significantly more prevalent in the negative and LSIL groups than in the HSIL group. This finding shows that although HPV52 is currently the most prevalent genotype in China, HPV16 and HPV58 may be the most important genotypes that cause HSIL.

03

Most HPV genotypes exhibited an increase in viral load corresponding to the severity of cervical lesions. However, the viral loads in the HSIL and LSIL groups were significantly higher than those in the negative control, and the difference was significant (p< .001). In addition to HPV18, HPV45, and HPV56, other high-risk HPV types showed significant differences in viral loads at different lesion levels. Except for HPV82, no significant differences were identified in the viral loads of medium- and low-risk HPV types at different lesion levels. The viral load of each HPV genotype was calculated by different methods and these findings indicate a correlation between HPV load and the degree of cervical lesions in multiple infections.

04

According to the ROC curve analysis (Table 4), the optimal cutoff for the viral loads of the 21 HPV types was 4.34 copies/10,000 cells (log10-transformed). At this cutoff value, the viral loads effectively detected HSIL, with an AUC of 0.76 (p < .05), a specificity (Sp) of 89.52%, and a sensitivity (Se) of 48.94%. For the viral loads of HPV16, the optimal cutoff value was 5.06 copies/10,000 cells (Se = 81.52%, Sp = 64.13%), with an AUC of 0.76 (p < .05). The optimal cutoff for HPV58 viral load was 4.31copies/10,000 cells (Se = 92.86%, Sp = 48.05%), with an AUC of 0.73(p < .05). Similarly, the HPV52 viral load had an appropriate cutoff value of 4.09 copies/10,000 cells (Se = 91.30%, Sp = 49.17%), leading to an AUC of 0.70 (p < .05).

05

We calculated the prevalence of HSIL across various HPV genotypes. HPV33 exhibited the highest HSIL prevalence, followed by HPV16. Although HPV73 had the highest prevalence of HSIL, the sample size was small, which limited its credibility.