深圳2 580例高危男性人乳头瘤状病毒基因型分析

目的 探讨深圳门诊高危男性患者人乳头瘤状病毒(HPV)感染的分布状况及基因型特点,为男性HPV感染的干预提供理论依据。方法 采用多重PCR结合反向斑点膜杂交的基因芯片技术对2012年3月～2014年6月在北京大学深圳医院收集的2 580例门诊疑似HPV感染的高危男性患者的生殖道脱落细胞进行23种HPV基因分型检测。结果 2 580例样本中HPV阳性1 164例,阳性率为45.12%, 其中低危型、高危型及高低危混合型感染的阳性率分别为51.03%(594/1 164),24.74%(288/1 164),24.23%(282/1 164)。低危型以HPV6(46.29%),HPV11(25.31%),HPV43(18.77%)和HPV42(9.62%)为常见亚型; 高危型以HPV16(16.58%),HPV52(12.59%),HPV33(10.60%),HPV58(7.98%)和HPV56(7.73%)为常见亚型。HPV感染以单一型感染为主(63.23%),二重及多重感染分别为10.48%和26.29%。年龄分层结果显示,不同年龄组HPV阳性率差异无统计学意义(χ²=6.98,P>0.05)。结论 深圳市高危男性HPV感染率较高,HPV6,11,43,42为主要的低危亚型,HPV16,52,33,58,56为主要的高危亚型,感染以单一亚型为主。     

HPV分型对宫颈癌患者放疗敏感性的影响

[目的]探讨宫颈癌患者中人乳头瘤病毒（HPV）分型与放疗敏感性的相关性.[方法]收集宫颈癌患者124例,检测患者HPV基因分型,采用放疗方法对入选患者进行盆腔外和腔内照射治疗,分析放疗敏感性与HPV分型的相关性.[结果]124例患者中,HPV阴性1例,在HPV阳性宫颈癌患者检出21种HPV基因型中,高危型常见类型为HPV16、HPV18和HPV58,低危型最常见类型为HPV6和HPV11.放疗不敏感患者中多重HPV感染所占90％,且HPV-18在化疗后未缓解患者中所占比率为80％.[结论]以HPV-18为主的HPV多重感染可能会影响宫颈癌患者放化疗敏感性.     

1268例慢性宫颈炎患者人乳头瘤病毒感染与基因型分布

目的调查慢性宫颈炎患者人乳头瘤病毒(HPV)感染及其基因型的流行分布状况。方法采用基因芯片技术对1268例慢性宫颈炎患者宫颈分泌物进行18种高危型HPV基因型和5种低危型HPV基因型的检测,分析其流行分布特点。结果 HPV阳性430例,占33.91%,高危型HPV感染占HPV阳性的82.93%,以HPV16为主,其次为HPV58、HPV18、HPV56。低危型感染占17.07%,以HPV11最高,其次为HPV6;单一感染占74.65%,重复感染占25.35%;高危型复合感染占66.06%,低危型+高危型复合感染占31.19%,单纯低危型的复合感染占2.75%。慢性宫颈炎患者HPV感染率以50岁组最高。结论慢性宫颈炎患者HPV的感染率较高,以高危型HPV基因型为主,临床应重视HPV感染的筛查、监测与治疗,防治宫颈癌。     

Synthetic genes: a tool for identifying human papillomavirus genotypes by hybridization and polymerase chain reaction-based assays

Obtaining positive polymerase chain reaction (PCR) controls for human papillomavirus (HPV) diagnostic tests has been difficult because of prevalence variation in different geographic regions of each high-risk viral type. Overlapping oligonucleotides were designed for HPV-18, HPV-31, HPV-45, and HPV-58 type-specific (TS) sequences. Synthetic HPV viral genes were constructed by 2-step assembly PCR for accurately diagnosing TS HPV infection.     

上海市普陀区人乳头瘤病毒分型检测及感染情况分析

目的：探讨上海普陀区妇女宫颈人乳头瘤病毒（HPV）感染亚型及其分布情况。方法采用聚合酶链反应（PCR）体外扩增和反向斑点杂交-基因芯片技术,检测2012年1月至2013年4月连续收集的共659例在本院门诊就诊妇女的宫颈脱落细胞标本。结果659例临床标本中共检出HPV感染340例,总检出率为51.6%。21种亚型中共检出20种亚型,其中HPV43低危亚型没有检出。感染率居前5位的基因型分别是16型为15.2%、CP8304型为10.2%、6型为8.9%、52型为8.9%、58型为8.4%。单一亚型感染比率为68%,多重感染为32%。20岁以下年龄组、20~29岁、30~39岁、40~49岁、50岁及以上年龄组HPV检出率分别为53.3%,60.7%,48.6%,36.6%及57.8%。高危亚型人群中,HPV主要检出亚型为16型、CP8304型、6型、52型及58型。结论29岁及以下年龄组检出高阳性率说明HPV感染有年轻化的倾向,其中16型、52型、58型等高危型HPV感染是本地区宫颈癌发病的主要原因。     

延安地区妇女HPV基因分型及临床相关研究

目的 对比分析延安市妇女人乳头瘤病毒(HPV)感染的基因型分布情况和临床特点,为宫颈癌筛查和防治提供数据支持。方法 采用导流杂交法对HPV感染型别进行基因分型,统计临床感染分布数据,并对其结果进行对比分析。结果 2 014例妇女中共检出HPV阳性者332例,阳性率16.48%,HPV高危型中HPV-16,52和53居于前三位,HPV低危型以HPV-81,6和44为主。阳性标本中单纯高危型感染219例,单纯低危型感染74例,混合感染39例,构成比分别为65.96%,22.29%和11.75%。HPV感染率随年龄增长呈现先增长后下降的趋势,各组比较差异有统计学意义(χ²=34.238,P<0.01)。不同职业人群HPV阳性检出率不同,各组比较差异有统计学意义(χ²=50.35,P<0.01)。不同地域人群HPV阳性检出率也存在差异,各组比较差异有统计学意义(χ²=12.084,P<0.05)。HPV感染结果中单一感染占比最高。结论 HPV感染率在不同人群、地域、年龄存在差异,进行HPV筛查对于宫颈癌的早期预防和治疗具有重要的意义。     

Type-specific detection of human papillomaviruses in a routine laboratory setting--improved sensitivity and specificity of PCR and sequence analysis compared to direct hybridisation.

Human papillomaviruses (HPV) are known to cause cervical dysplasia and cervical carcinoma. We used a 3-step PCR protocol that allows rapid type-specific HPV testing in a routine laboratory setting: HPV-16-positive samples were determined using a specific LightCycler PCR; HPV-16-negative samples were amplified by nested PCR and typed by sequence analysis. During a period of 7 months, 1275 PCR-based HPV tests were performed. Of the 1275 samples, 829 samples tested negative for HPV and 446 tested positive, including 124 positives found in the initial HPV-16-specific LightCycler assay. Sequence analysis of 132 samples detected 18 HPV types that are not included in the widely used Hybrid Capture II assay. For comparison, the first 100 cervical specimens were tested in parallel using PCR and direct hybridisation (Hybrid Capture II assay). PCR detected HPV DNA in 23 samples that tested negative in the Hybrid Capture assay. Four out of 37 samples that tested positive for HPV in the Hybrid Capture test may be false positives, because sequence analysis detected HPV types not included in the probe mixtures. As rare and novel HPV types may also confer an oncogenic risk, highly sensitive and specific PCR assays will help in understanding cervical HPV infection and cervical cancer of unknown causes.     

Sentinel-base DNA genotyping using multiple sequencing primers for high-risk human papillomaviruses

Despite the various technologies in place for genotyping human papillomaviruses (HPV), clinical use and clinical research demand a method that is fast, more reliable and cost-effective. The technology described here represents a breakthrough development in that direction. By combining the method of multiple sequencing primers with DNA sequencing, we have developed a rapid assay for genotyping HPV that relies on the identification of a single, type-specific 'sentinel' base. As described here, the prototype assay has been developed to recognize the 12 most high-risk HPV types (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59) and is capable of recognizing and simultaneously genotyping multiple HPV co-infections. By providing sequence information on multiple HPV infections, this method eliminates the need for labor- and cost-intensive PCR cloning. These proof-of-concept studies establish the assay to be accurate, reliable, rapid, flexible, and cost-effective, providing evidence of the feasibility this technique for use in clinical settings.     

A novel method for multiplex genotyping in a single reactor using GTPlex-PyroSeq: genotyping HPV as a prototype

Herein, we describe a novel multiplex genotyping method, GTPlex-PyroSeq. This method consists of two phases: multiplex PCR followed by a single reaction of pyrosequencing. This study demonstrates how GTPlex-PyroSeq can be adapted for the determination of multiple human papillomavirus (HPV) genotypes. A biotinylated consensus primer, GP6+, and 15 high-risk HPV type-specific primers are used for multiplex PCR. Each type-specific primer has a 5'-tag unique ID sequence connected to a pyrosequencing primer binding region. The unique ID sequence is composed of three parts: i) a single nucleotide ID representing a specific genotype; ii) a sign post; and iii) an end mark. This design allows multiple genotype determination under an ID sequence-dependent nucleotide dispensation order during pyrosequencing. Following initial studies using HPV plasmids and cell lines, we evaluated the clinical utility and effectiveness by comparing our assay with direct sequencing and HPV DNA chip analysis of 80 samples from high-risk, HPV-positive patients. We found in single-type infections, 100% concordance with direct sequencing (70 of 80 perfect matches) and 97.5% concordance with HPV DNA chip data (50 of 80 perfect matches). Additionally, our system was superior to direct sequencing in detection of multiple infections (12 of 80), with a limit of detection of 100 copies. The scalability of this multiplex system, with its open-platform design and ability to use various sample types, makes the GTPlex applicable for use in multiple settings.     

人乳头瘤病毒（HPV）6／11、16／18型检测及结果分析的临床意义

目的用实时荧光定量PCR法检测低危型人乳头状瘤病毒HPv（如6／11型）和高危型人乳头状瘤病毒HPV（如16／18型）,探讨其在宫颈癌防治方面的意义。方法采用FQ．PCR方法检测624例不同年龄组患者感染HPV6／11、HPV16／18型情况。结果624例标本中HPV6／11、HPV16／18检测的阳性率分别为26-3％,8．3％,低危型远高于高危型（P〈0．01）。HPV6／11、HPV16／18阳性患者的年龄分布不同,年龄段的感染情况也不同,HPV6／11型感染主要集中在20～40岁,HPV16／18型感染者年龄均在41～50岁。结论荧光定量PCR检测低危型HPV和高危型HPV,方法简便准确,快速。     

Molecular evaluation of the prevalence of oncogenic human papillomavirus genotypes in cervical acetowhite lesions

The actual prevalence of cancer-related human papillomavirus (HPV) genotypes in cervical acetowhite lesions has not been established. In this work, the presence of oncogenic types of HPV in cervical acetowhite tissue was evaluated by molecular means. The presence of HPV DNA was determined in a group of women with and without cervical acetowhite lesions by a polymerase chain reaction (PCR) using the MY09/MY11 primers. The presence of 13 oncogenic HPV types was evaluated using the Hybrid Capture II test, and the prevalence of HPV type 16 (HPV16) was studied using an HPV16-specific PCR. HPV DNA was detected in 85.9% patients with acetowhite lesions; oncogenic HPV types were found in 83.7% of them; HPV16 was identified in 51.1% of the cases. HPV DNA was detected in 87.3% of the patients without acetowhite changes. Interestingly only 16.8% were infected by oncogenic genotypes and 2.5% were positive for the presence of HPV16. In conclusion, subclinical infection by oncogenic HPV genotypes is associated with the presence of acetowhite cervical tissue (p < 0.0005). Therefore, women showing acetowhite lesions might be at risk of developing cervical cancer.     

Detection of oral HPV infection – Comparison of two different specimen collection methods and two HPV detection methods

Very little is known about the natural history of oral HPV infection. Several different methods exist to collect oral specimens and detect HPV, but their respective performance characteristics are unknown. We compared two different methods for oral specimen collection (oral saline rinse and commercial saliva kit) from 96 individuals and then analyzed the samples for HPV by two different PCR detection methods (single GP5+/6+ PCR and nested MY09/11 and GP5+/6+ PCR). For the oral rinse samples, the oral HPV prevalence was 10.4% (GP+ PCR; 10% repeatability) vs 11.5% (nested PCR method; 100% repeatability). For the commercial saliva kit samples, the prevalences were 3.1% vs 16.7% with the GP+ PCR vs the nested PCR method (repeatability 100% for both detection methods). Overall the agreement was fair or poor between samples and methods (kappa 0.06–0.36). Standardizing methods of oral sample collection and HPV detection would ensure comparability between future oral HPV studies.     

重庆地区人乳头瘤病毒各亚型感染情况的临床分析

目的了解重庆地区人乳头瘤病毒(HPV)各亚型感染的分布情况,为重庆地区HPV分子流行病学研究提供依据。方法采用导流杂交技术,同时检测21种HPV亚型,其中高危型HPV 13种,低危型HPV 5种和中国人群常见型HPV 3种。结果 3 884例临床标本共检出阳性1 144例(29.45%,1 144/3 884),单一HPV亚型感染750例(19.31%,750/3 884),复合感染394例(10.14%,394/3 884),其中高危型HPV 515例(13.26%,515/3 884,不含复合感染);21种HPV亚型均被检出,其中HPV52、16、58、33、68是最常见的高危型;58例男性标本中检出31例(53.45%)阳性,主要为复合型和HPV6型。结论 HPV各亚型检测对于HPV感染的诊断、治疗及宫颈癌的早期干预和预防具有重要意义。     

人乳头瘤病毒基因分型方法的建立和应用

目的 建立一种简便快捷、灵敏度高、经济实用的人乳头瘤病毒(HPV)基因分型的低密度基因芯片技术,并对该方法进行评价.方法 用低密度基因芯片技术对355例疑似HPV感染女患者检测HPV并分型;用杂交捕获Ⅱ代(HCⅡ)法进行评价.并用该法对珠江三角洲地区的730例标本进行HPV分型检测.结果 355例疑似样本中,低密度基因芯片技术和HCⅡ法分别检出211例(59.4%)和222例(62.5%)阳性标本,符合率达94.1%(334/355).低密度基因芯片技术共检测出15种常见高危型和5种低危型,其中16、52、58、56型检出率较高.结论 低密度基因芯片技术能具体分型和检测混合感染,HPV检测与细胞学检测结合.对宫颈癌筛查有重要意义.     

新疆地区妇女宫颈病变组织中HPV感染与分型研究

目的探讨人乳头瘤状病毒（human papillomavirus,HPV）型别在宫颈病变中分布情况。方法采用基因芯片技术对239例宫颈癌前病变或宫颈鳞癌患者的石蜡组织标本进行23种HPV基因型别检测。结果检测出20种基因型,HPV总感染率为90．0％（215／239）;单一感染中高危型HPV16感染率为84．2％（117／139）,低危型中HPV11感染检出1例;多重感染率为35．3％（76／215）,64例为HPV16与其他高危病毒的复合感染,5例为HPV16与低危病毒（6,11,43）的复合感染,7例为低危病毒与低危病毒的复合感染。结论高危型HPV16感染是新疆地区妇女宫颈病变中常见亚型,多重感染可能与宫颈病变进展无关。     

Low-density addressable array for the detection and typing of the human papillomavirus

We have developed a low-density DNA array for the detection and typing of human papillomavirus (HPV) DNA. The gene chemistry strategy involves using a combination of the polymerase chain reaction (PCR) with the consensus oligonucleotide primers MY09/MY11 followed by a ligase detection reaction (LDR). Fluorochrome-labeled HPV-specific primers are joined to a common primer modified with a unique anchoring sequence called a zip code on its 3' end. The result is a series of 60-70 base pair and single-stranded ligation products that are then hybridized to their respective zip code complements affixed to glass slide based arrays. Nine separate zip codes were assigned, one for each HPV type (6,11,16,18, 31, 33, 35, and 53) and one for a beta-globin internal control marker. Two additional zip-codes were reserved for a pair of consensus HPV LDR products: the cLDR1 and cLDR2 primers hybridize to a conserved sequence within the HPV L1 open reading frame internal to the MY09/MY11 fragment. These consensus primers were shown to detect over 40 different HPV types. The purpose of this study was to evaluate the analytic performance of this low-density microarray based assay for HPV, as well as to introduce our simplified read-out instrumentation, shown here to be a low cost and highly efficient way to detect and genotype HPV for clinical testing.     

人乳头瘤病毒6/11、16/18型的检测及临床意义

目的 用实时荧光定量聚合酶链反应(PCR)法检测低危型人乳头瘤病毒(HPV)(如6/11)和高危型HPV(如16/18),探讨其在宫颈癌防治方面的意义.方法 采用实时荧光定量PCR对2 162例不同年龄组患者进行HPV-DNA(6/11、16/18)检测.结果 低危型HPV的感染率为11.78%,高危型 HPV的感染率为11.40%,二者的感染率差别不大.结论 HPV为泌尿生殖道感染的重要病原,对妇女进行高危及低危型HPV-DNA检测对早期诊断、治疗具有重要的指导意义,高危型HPV在宫颈癌早期病变的筛查中具有风险提示作用.     

宫颈上皮内瘤变及鳞癌中人乳头瘤病毒不同亚型分布及多重感染的研究

目的探讨宫颈上皮内瘤变及宫颈癌中不同HPV亚型分布及多重感染情况的临床意义。方法选取2019年7月至2020年6月四川大学华西第二医院病理科诊断为宫颈上皮内瘤变(CIN)Ⅰ、Ⅱ、Ⅲ级及宫颈癌的四川地区病例384例,其中CINⅠ、CINⅡ、CINⅢ分别为77、50、166例,宫颈癌91例。运用返向点杂交法检测其HPV感染情况,统计不同HPV亚型在CIN及宫颈癌中的分布及多重感染情况。结果 384例患者总的HPV感染率为95.80%,各组病变均以高危型感染为主。高危型HPV在CINⅠ、CINⅡ、CINⅢ及宫颈癌中感染占比逐渐升高,分别为88.52%、92.96%、95.05%、97.96%;而低危型HPV在各组间感染占比则逐渐降低,分别从11.48%、7.04%、4.95%到2.04%,各组病变中高、低危型HPV组间差别有统计学意义(P=0.028)。总病例中最常见的HPV感染亚型为16、58、52、33、18型,此五种亚型在宫颈癌中感染率达96.7%,其中HPV16型感染率高达71.43%。感染病例中的多重感染率为25.54%。CINⅠ、CINⅡ、CINⅢ及宫颈癌中多重感染占比分别为50...     

贵州地区8102例就诊妇女21种HPV亚型检测结果分析

目的：分析贵州地区8102例就诊妇女人乳头瘤病毒（HPV）的感染情况、亚型分布特点。方法采用填写调查表方式,对受检妇女的一般情况、宫颈病变病理分级及HPV感染的高危因素进行调查;采用凯普核酸分子导流杂交基因芯片技术进行21种HPVDNA亚型分析。结果8102例就诊妇女21种HPV型别全部检出,阳性率为34．47％,高危型HPV阳性率29．60％,低危型为5．20％,高危型检出率的前3位是HPV16、52和58亚型,低危型以HPV11和6亚型为主。3040例宫颈炎患者中检出944例HPV阳性,阳性率为31．05％,高危型HPV阳性率26．64％,低危型HPV阳性率3．68％,高危型中检出率前6位的是HPV16、52、58、33、18和31亚型,低危型以11和6亚型为主;433例宫颈上皮内瘤变（CIN）Ⅰ～Ⅲ患者共检出240例HPV高危型阳性,阳性率为55．43％,检出率前6位是16、58、52、33、18和51亚型;249例宫颈癌患者共检出196例HPV高危型阳性,阳性率78．71％,检出率前6位是16、58、18、52、33和31亚型;宫颈炎、CINⅠ～Ⅲ和宫颈癌组均以单一型感染为主。受检者不同初次性生活年龄、伴侣个数、妊娠次数、生育次数的HPV感染率差异无统计学意义（P＞0．05）。结论贵州地区就诊妇女HPV感染率较高,16、52、58和18亚型为较常见高危亚型,6和11亚型为较常见低危亚型。