Epidemiological Characteristics And Genomic Diversity Of HPV11 In Patients With Condyloma Acuminatum Based On Sentinel Hospitals

Condyloma acuminatum(CA)is a common sexually transmitted disease(STD)caused by human papillomavirus(HPV)infection.Although CA is the most common benign lesion in the anogenital region,it often recedes and requires repeated treatment.The treatment process is painful and expensive,which brings severe psychological stress and economic burden to patients and reduces their quality of life.More than 200 HPV genotypes have been identified,classified as high-risk(HR)and low-risk(LR)based on their carcinogenic risk,and at least 40 genotypes are associated with anogenital infections,with 90%of CA primarily caused by HPV6 and HPV 11 infections.As one of the important low-risk types of HPV causing CA,HPV11 has also been found to be associated with malignant transformation of anal squamous cell carcinoma and laryngeal papilloma.At present,there are only foreign studies on the characteristics of the whole genome sequence of HPV11,and no such studies have been reported in mainland China.The homology and nucleotide variation of HPV 11 virus strains in foreign countries are not clear.As a province with a high number of reported CA cases,Shandong province also has very limited information on the epidemiological characteristics of CA patients and the distribution of HPV genotypes,especially the lack of understanding of the psychological and life quality of CA patients.Therefore,this study analyzed the epidemiological characteristics of CA patients in Shandong province through case investigation and laboratory tests in three sentinel hospitals in two regions from August 2019 to June 2021.Single positive samples of HPV 11 were sequenced by next-generation sequencing to explore the genomic diversity and prevalence of the whole HPV11 genome.The results of this study clarified the epidemiological characteristics of CA patients in Shandong Province,and also provided molecular basis for the study of the association between HPV 11 genomic diversity and CA,which has guiding significance for the prevention and treatment of CA in Shandong Province.Objective1.To master the disease characteristics of CA patients in Shandong Province,and provide data support for the formulation of CA prevention and treatment strategies in Shandong Province;2.Understand the psychology and quality of life of CA patients,and provide reference for clinical psychological intervention for CA patients;3.To analyze the whole genome characteristics of HPV 11 infection in CA patients,and to provide molecular basis for studying the relationship between genomic diversity of HPV11 and CA.Methods1.Case investigation of CA patients:A one-to-one questionnaire survey was conducted on CA patients who visited three sentinel hospitals in two regions of Shandong Province from August 2019 to June 2021.The contents of the questionnaire included the demographic characteristics,sexual behavior characteristics,clinical characteristics,knowledge of CA,and psychological and quality of life evaluation scales of CA patients.The psychological and quality of life survey included four scales,SDS,CECA10,DLQI and ED-5D-5L.The questionnaires were entered through Epidata software,an Excel database was created and statistical analysis was carried out using SPSS 23.0 software.2.HPV typing and sequencing:samples were collected from patients who agreed to undergo HPV typing testing after receiving the questionnaire.Nucleic acids from exfoliated cell samples were extracted,amplified by PCR,and HPV genotypes were determined using PCR-reverse dot blot hybridization.Samples with a single positive HPV11 typing result were selected,and the whole genome sequence of HPV 11 was amplified in fragments,and sequenced by next-generation sequencing technology.3.HPV 11 genomic diversity analysis:The sequencing results were spliced and sorted using CLC Genomics Workbench v12.0.2.The obtained sequences were globally aligned using MAFFT v7.490.PhyML v3.0 was used to construct a phylogenetic tree based on the maximum likelihood method.MEGA X was used to calculate the nucleotide differences between pairwise sequences of HPV 11 whole genome,DnaSP v6 detected HPV 11 nucleotide variation sites.Results1.From August 2019 to June 2021,a total of 1201 CA patients were recruited for case investigation,and 1185 valid questionnaires were received,including 870 males and 315 females.The average age was 34.85± 13.60,and male patients were older than female patients(P<0.01).Most of the patients came from cities(69.7%),60.3%of the patients were married,51.3%of the patients had a college education or above,and 57.0%were freelancers and corporate employees.2.Among CA patients,89.1%had heterosexual sex,the average age at first sexual intercourse was 21.2±3.01,and 57.8%of the patients had first sexual intercourse younger than legal marriageable age.Male patients had more sex than female patients in total.partner(P<0.001).Only 56.8%of patients knew CA,and 41.4%knew that CA was infected by HPV.52.0%and 32.7%of the patients were aware of HPV transmission routes and prevention methods,respectively.Females were more aware of HPV knowledge than males(P<0.001).3.There were 411 patients with recurrence,and the recurrence rate was 34.7%.13.5%of the patients’ sexual partners also had CA,and 34 patients were co-infected with other sexually transmitted diseases.The main infection sites of male patients were the coronary sulcus,foreskin and perianal,and the main infection sites of female patients were the urethra,vaginal opening,labia majora and perineum.46.5%of patients had multiple site infections.4.Among the 880 CA patients who received HPV typing,the overall HPV infection rate was 91.4%.The most common HPV genotypes were HPV 6(57.8%),HPV 11(37.2%),HPV 16(13.7%)and HPV 42(10.3%).In these cases,low-risk HPV infection predominated,with an infection rate of 91.3%,and 87.4%were HPV6 and/or HPV11 infection.Compared with women,men had a higher rate of low-risk HPV infection(P<0.001).High-risk HPV infection was also detected in 53.5%of the patients.Among infection types,single infection was more common in male patients(P<0.001),while multiple infections were more common in female patients(P<0.001).Among specific types,the infection rate of HPV 45,HPV 56 and HPV 58 in females was higher than that in males(P<0.05).5.A total of 1161 valid psychological and life quality questionnaires were received,of which 73.6%were male and 26.4%were female.The mean standardized score of SDS was 56.54±8.16,and 72.5%of the patients had depressive symptoms.The standardized score of CECA10 scale was 48.82±24.49.In terms of psychological and emotional dimensions,the items with the lowest scores were "I am afraid that the lesions will not disappear" and "I feel worried when I have sex" respectively.The average score of DLQI scale was 6.97±6.23,and 78.4%patients’ quality of life was affected.EQ-5D-5L scale showed that the health utility value of CA patients was 0.894±0.152.Anxiety or depression had the most problems,accounting for 53.5%,of which 3.3%had extremely serious problems.6.A total of 24 HPV11 whole genome sequences were obtained by next-generation sequencing.Phylogenetic analysis was performed with 97 sequences from other regions of the world in GenBank,It was found that all the sequences obtained in this study were from A lineage and could be further divided into A1 sublineage(n=7),A2 sublineage(n=15),A3 sublineage(n=1)and A4 sublineage(n=1).The A2 sublineage was the main branch of HPV11 genome sequence in this study,accounting for 62.5%.The nucleotide difference of 121 sequences was 0-1.50%.Compared with the standard HPV 11 sequence,a total of 105 nucleotide mutants were identified in the measured sequence.Analysis showed that E1,L1 and L2 were relatively highly variable regions of HPV11.The A2 sublineages have more genomic diversity than the A1 sublineages.Conclusions:1.Most of the patients with CA are sexually active,have previous experience of risky sex,and lack of understanding of CA,resulting in a high recurrence rate.Compared with other regions,the prevalence rate of HPV42 in CA patients in Shandong province was higher.This suggests the need for better health education for CA patients,as well as regular monitoring of non-vaccine covered HPV types.2.Most CA patients have depressive symptoms and their quality of life is greatly affected.Although the patients’ physical health and social function are generally good,clinicians are advised to strengthen mental health interventions for patients to help patients alleviate psychological problems and improve their quality of life.3.Nearly 40%of CA patients in Shandong province were infected with HPV11.The whole genome sequence of HPV11 belonged to A lineage,and the sublineage was mainly A2.Nucleotide differences were slightly higher(0～1.50%)compared with the rest of the world.The mutation sites of A1 and A2 sublineages were mainly located in L1 and L2 coding regions,and the A2 sublineages had higher genomic diversity than the A1 sublineages.