研究目的

    本研究旨在评估研究中适用于男性肛门-生殖器标本HPV基因分型的方法，并基于此分型方法，分析石家庄市男性普通人群肛门-生殖器各部位HPV感染率、型别分布、多重感染情况，探究影响肛门-生殖器各个部位HPV感染的危险因素。

研究方法

1. 基于男性肛门-生殖器标本的不同HPV检测方法的性能评估

    370份肛门-生殖器细胞学标本（198例外生殖器标本、128例会阴/肛周标本和44例肛门标本），以及58例病变程度不同的由福尔马林固定、石蜡包埋的肛门-生殖器组织学标本（8例会阴/肛周标本和50例肛门标本）被纳入研究。分别使用LiPA-25和SureX-25两种HPV基因分型检测方法评估标本中20个共有HPV型别检出情况，包括14个高危型HPV（HPV 16、18、31、33、35、39、45、51、52、56、58、59、66和68/73）和6个低危型HPV（HPV 6、11、42、43、44、53）。使用一致率和kappa值评估两种检测方法检出HPV型别的一致性。

2. 石家庄市男性普通人群肛门-生殖器部位HPV感染型别分布及影响因素研究

    本研究为横断面研究，于2022年1-4月在河北省石家庄市招募490名18~45岁男性受试者，通过问卷调查收集受试者主要社会人口学特征、性行为方式等个人信息。每位受试者分别从外生殖器、会阴/肛周及肛门采集脱落细胞学标本，使用SureX-25 HPV检测方法，对符合检测标准的标本进行HPV DNA分型检测。采用构成比和HPV感染率进行统计学描述；用McNemar检验和kappa值比较不同解剖部位HPV感染的一致性和差异性；采用单因素c²检验和多因素logistic回归分析探究影响男性不同解剖部位的HPV感染率的影响因素。

研究结果

1. 基于男性肛门-生殖器标本的不同HPV检测方法的性能评估

    SureX-25和LiPA-25两种检测方法检测肛门-生殖器细胞学标本中的任一20种HPV型别和任一高危型HPV的一致性极好，kappa值分别为0.825和0.821。低危型HPV 6（kappa=0.914）和高危型HPV 33（kappa=1.000）、HPV 35（kappa=1.000）、HPV 51（kappa=0.973）和 HPV 52（kappa=0.925）的一致性极高。对HPV 11（kappa=0.888）、HPV 16（kappa=0.861）、HPV 66（kappa=0.842）和HPV 68/73型（kappa=0.858）表现出了较好的一致性。此外，与LiPA-25相比，SureX-25对特定型别如HPV 56、HPV 58、HPV 59和HPV 44的检出率更高（P<0.05）。分部位来看，两种检测方法检测198例外生殖器标本和128例会阴/肛周标本中，任一高危型HPV的一致性均极好（均kappa >0.800）。在58份组织学标本中，两种检测方法检出高危型HPV的一致性极高（kappa=0.825），对任一HPV型别（kappa=0.676）和低危型HPV（kappa=0.726）检出率一致性较好。SureX-25和LiPA-25在检测高危型HPV 16、HPV 45、HPV 51、HPV 52和HPV 58上具有极高的一致性（均kappa＞0.800）；对于低危型别，两种检测方法均有较高一致性（均kappa＞0.600）。

2. 石家庄市男性普通人群肛门-生殖器部位HPV感染型别分布及影响因素研究

    石家庄市男性普通人群总体HPV感染率为29.08%（107/368），高危型HPV（HPV 16/18/31/33/35/39/45/51/52/56/58/59/66/68）和低危型HPV（HPV 6/11/42/43/44/81/83）感染率分别为23.64%（87/368）和13.04%（48/368）。此外，共获得394例外生殖器、392例会阴/肛周和481例肛门符合分型检测标准的标本。外生殖器、会阴/肛周和肛门部位HPV感染率分别为23.10%（91/394）、15.31%（60/392）和2.08%（10/481），差异有统计学意义（P<0.05）。高危型HPV在上述3个部位的感染率分别是18.02%（71/394）、11.22%（44/392）和1.46%（7/481），低危型HPV的感染率则分别为9.64%（38/394）、5.87%（23/392）和0.62%（3/481）。外生殖器部位最常见的HPV型别为HPV 52、51、56、59、68，会阴/肛周部位为HPV 44、66、56和58，而在肛门最常见的型别则为HPV 43和45。肛门-生殖器部位均以单一感染为主（均P <0.05）。不同部位间HPV型别一致性较差。有多名性伴侣的男性外生殖器或会阴/肛周部位HPV感染风险相对更高。

研究结论

1. 基于男性肛门-生殖器标本的不同HPV检测方法的性能评估

    SureX-25在检测男性肛门-生殖器标本中的HPV基因分型方面表现出与 LiPA-25相当的性能，SureX-25对于高危型HPV更灵敏。SureX-25还在标本质量控制、操作简便性和低成本效益方面具有显著优势，为男性人群检测和临床实验的HPV基因分型方法提供了更多选择。

2. 石家庄市男性普通人群肛门-生殖器部位HPV感染型别分布及影响因素研究

    石家庄市男性HPV感染率较高，尤其是以外生殖器部位为主；不同部位之间HPV感染率存在显著差异。有多名性伴侣的男性，其外生殖器和会阴/肛周部位感染HPV的概率更高。应制定合理有效的预防措施降低男性HPV的感染，从而有助于降低两性HPV感染相关疾病的发生。

Objectives

The aim of this study was to evaluate HPV genotyping assays applicable to male anogenital specimens in epidemiological studies, to provide a methodological basis for accurate detection of HPV infection in men, and based on this to analyse the prevalence of HPV infections in different anogenital sites of the male general population of Shijiazhuang, and to elucidate the risk factors for HPV infections in different anogenital region.

Methods

1. Performance evaluation of different HPV assays for male anogenital specimens

370 anogenital cell specimens (198 the external genital, 128 perineal/perianal, and 44 anal specimens, and 58 formalin-fixed, paraffin-embedded anogenital tissue specimens (8 perineal/perianal and 50 anal specimens) with varying grades of lesion were selected for the study. The agreement of 20 common HPV types, including 14 high risk HPV (HR-HPV) types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68/73; 6 low risk HPV (LR-HPV) types: 6, 11, 42, 43, 44, and 53 between the SureX-25 and LiPA-25 assays was assessed using agreement rates and Cohen's kappa (κ).

2. The prevalence and associated factors of human papillomavirus infection in different anogenital sites among men in Hebei province

A cross-sectional study was conducted in Shijiazhuang, Hebei Province, and 490 healthy men aged 18-45 years were recruited from January to April, 2022. The main demographic, sexual behavior, and other individual characteristics were collected by questionnaire. Swab specimens were obtained from the external genital (penis/glans penis/coronal sulcus, PGS), perianal/ perineal (PA), and anal sites of each participant. HPV genotyping was performed by SureX-25 assay. Statistical descriptions were performed using constitutive ratios and HPV infection rates; McNemar's test and Cohen's kappa values were used to compare the consistency and variability of HPV infections at different anogenital sites; and Pearson's Chi-squared test and multivariate logistic regression analyses were used to explore the associated factors of the prevalence of HPV infections in different anogenital sites among men.

Results

1. Performance evaluation of different HPV assays for male anogenital specimens

The concordance of SureX-25 and LiPA-25 assays for detecting HPV in anogenital swab samples was comparable for any 20 HPV types and HR-HPV, with κ values of 0.825 and 0.821, respectively. High agreement was observed for low-risk HPV type 6 (κ=0.914) and high-risk types 33 (κ=1.000), 35 (κ=1.000), 51 (κ=0.973), and 52 (κ=0.925). Good concordance was also noted for types 11 (κ=0.888), 16 (κ=0.861), 66 (κ=0.842), and 68/73 (κ=0.858). Additionally, SureX-25 demonstrated higher detection rates for specific types such as HPV56, HPV58, HPV59 and HPV 44(P<0.05 each). By site, there was high agreement (κ>0.800 each) for either HR-HPV in 198 PGS and 128 PA specimens tested by both assays. In 58 tissue samples, high concordance was observed for the detection of HR-HPV (κ=0.825). Detection rates for any HPV and LR-HPV were similar between the assays. The SureX-25 assays also showed perfect agreement with the LiPA-25 assay for HPV16, HPV 45, HPV 51, HPV 52 and HPV58 (κ > 0.800 each). For LR-HPV, there was a good concordance between the two assays (κ > 0.600 each).

2. The prevalence and associated factors of human papillomavirus infection in different anogenital sites among men in Hebei province

The overall HPV infection rate of the male general population in Shijiazhuang was 29.08% (107/368), and the infection rates of HR-HPV (HPV16/18/31/33/35/39/45/51/

52/56/58/59/66/68) and LR-HPV(HPV6/11/42/43/44/81/83) were 23.64% (87/368) and 13.04% (48/368), respectively. Valid HPV genotyping was completed for 394 PGS, 392 PA, and 481 anal specimens. The prevalence of HPV infection was 23.10% (91/394) in PGS, 15.31% (60/392) in PA, and 2.08% (10/481) in the anal site. The prevalence of HR-HPV in PGS, PA, and anal sites was 18.02% (71/394), 11.22% (44/392), and 1.46% (7/481), respectively, while the prevalence of LR-HPV was 9.64% (38/394), 5.87% (23/392) and 0.62% (3/481), respectively. The most common HPV types were HPV52, 51, 56, 59 and 68 in PGS; HPV44, 66, 56, and 58 in PA; and HPV43 and 45 in the anal site. Single HPV infection was dominant among all sites (P <0.05). Poor concordance of HPV types was observed between different sites. Individuals with multiple sexual partners had a significantly higher risk of HPV infection in PGS and PA.

Conclusions

1. Performance evaluation of different HPV assays for male anogenital specimens

SureX-25 demonstrated comparable performance to the LiPA-25 assay in male anogenital specimens, particularly in cell specimens. SureX-25 offers good advantages in terms of samples quality control, ease of use, and cost effectiveness, providing more options for HPV genotyping methods suitable for use in male population screening and in clinical trials.

2. The prevalence and associated factors of human papillomavirus infection in different anogenital sites among men in Hebei province

Men in Shijiazhuang had a high prevalence of HPV infection, especially predominantly in PGS. There was a significant difference in the HPV infection rates between different anogenital sites. Having multiple sexual partners is a risk factor for HPV infection of PGS and PA in men. Reasonable and effective preventive measures should be developed to reduce HPV infection in the male anogenital sites, thus contributing to the reduction of HPV infection-related diseases in both men and women and promoting the prevention.