PCR-寡核苷酸流芯片用于人乳头状瘤病毒分型检测

目的应用PCR-寡核苷酸微流芯片检测方法进行人乳头状瘤病毒(HPV)分型,研究子宫颈上皮内瘤变(CIN)与HPV感染型别的关系.方法利用简并引物对所有HPV L1保守区进行扩增,再通过寡核苷酸微流芯片上特异性探针对CIN进行HPV分型.结果41例CIN病人中22例(54%)HPV检测为阳性,低级CIN1组高危HPV感染仅16%,CIN2组高危HPV型感染50%,高级CIN3组高危HPV型感染75%;与DNA测序方法比较PCR-寡核苷酸微流芯片检测方法具有较好的灵敏度.结论随着CIN级别的升高,高危HPV型感染比例增加,寡核苷酸微流芯片检测能区分多种HPV型别,可用于临床宫颈上皮内瘤变的HPV筛查.     

扩增人肝型磷酸果糖激酶基因对唐氏综合征进行定量基因诊断的研究

目的　扩增位于唐氏综合征关键区域的人肝型磷酸果糖激酶基因 (PFKL基因 ) ,对唐氏综合征进行定量基因诊断。方法　采用定量PCR—微孔板杂交检测PCR产物量的方法 ,对 2 6例唐氏综合征患者及 2 78例正常人外周血DNA标本 ,扩增PFKL基因 ,扩增片段长度为 185bp ;另外 ,将人肌型磷酸果糖激酶基因 (PFKM基因 )作为内参照同时扩增 ,扩增片段长度为 36 5bp ,定量PCR产物用微孔板杂交检测。 结果　 2 6例患者 (包括 1例易位型 )PFKM与PFKL扩增产物的OD值比值介于 0 4 0～ 0 6 0之间 ,平均值为 0 5 1;正常人扩增产物的OD值比值介于 0 80～ 1 2 0之间 ,平均值为 1 12 ,两者比较差异显著 (P <0 0 0 1)。对 2 78例正常人进行同样基因扩增和检测无一例假阳性 ,所得结果与染色体核型分析结果完全符合。结论　定量PCR—微孔板杂交检测PFKL基因拷贝数的方法 ,简便、快速、特异 ,可用于唐氏综合征基因诊断     

扩增大肝型磷酸果糖激酶基因对唐氏综合征进行定量基因诊断的研究

目的：扩增位于唐氏综合征关键区域的大肝型磷酸果糖激酶基因（PFKL基因），对唐氏综合征进行定量基因诊断，方法：采用定量PCR－微孔板杂交检测PCR产物量的方法，对26例唐氏综合征患者及278例正常人外周血DNA标本，扩增PFKL基因，扩增片段长度为185bp；另外，将人肌型磷酸糖激酶基因（PFKM基因）作为内参照同时扩增，扩增片段长度为365bp，定量PCR产物用微孔板杂交检测。结果：26例患者（包括1例易位型）PFKM与PFKL扩增产物的OD值比值介于0．40－0．60之间，平均值为0．51；正常人扩增产物的OD值比值介于0．80－1．20之间，平均值为1．12，两者比较差异显著（P＜0．001）。对278例正常人进行同样基因扩增和检测无一例假阳性，所得结果与染色体核型分析结果完全符合。结论：定量PCR－微孔板杂交检测PFKL基因拷贝数的方法，简便，快速，特异，可用于唐氏综合征基因诊断。     

导流杂交与杂交捕获二代用于生殖道人乳头状瘤病毒感染诊断的比较研究

目的评价导流杂交在女性生殖道人乳头状瘤病毒（HPV）感染检测中的实用价值.方法对2004-08-2005-01就诊于中日友好医院妇产科的310例女性宫颈脱落细胞标本,用导流杂交法进行21种HPV亚型的分型检测,同时用杂交捕获二代（HC-Ⅱ）进行13种高危型HPV检测.比较两种方法13种高危型HPV检测结果的符合情况,结果不一致标本进行基因测序.结果两种方法检测结果一致性良好,不同病变组Kappa指数均＞0.4.测序结果60.0%与导流杂交法符合.结论导流杂交检测效果与HC-Ⅱ具有可比性,且能确切分型,是较理想的HPV检测方法.     

SOX1及PAX1基因甲基化检测在子宫颈癌筛查中的价值研究

目的 探讨不同程度子宫颈病变中SOX1及PAX1基因甲基化水平,以期为子宫颈癌筛查方案提供参考依据。方法 选取滦州市人民医院2021年9～12月收治的133例高危型人乳头瘤病毒（HPV）阳性患者,应用特异性多重PCR荧光-探针法对子宫颈脱落细胞中SOX1及PAX1基因的甲基化水平进行检测,研究其与DNA倍体分析、HPV分型检测、薄层液基细胞学检测（TCT）及组织病理学结果之间的关系。结果 甲基化检测子宫颈高度病变/子宫颈癌的阳性率为88.9%,显著高于DNA倍体分析（44.4%）;甲基化检测不同型别HPV感染的阳性率由高到低依次为：HPV33(50.0%)、HPV16(32.7%)、HPV58(30.8%)、HPV18(11.1%)、HPV52(10.5%);在甲基化与TCT检测结果比较中,甲基化检测对正常子宫颈组的阳性率更低（13.3%）;在综合诊断性能对比中,甲基化检测相比TCT、DNA倍体分析技术显示出了更高的特异度（84.35%）、阳性预测值（47.06%）与准确度（84.96%）。结论 SOX1及PAX1基因甲基化检测可用于子宫颈癌初筛检测的辅助诊断,对子宫颈癌前病变进行精准...     

生殖道人乳头瘤病毒感染与宫颈病变、宫颈癌相关性研究

目的 了解生殖道人乳头瘤病毒感染与宫颈病变、宫颈癌的关系，以便早期发现和治疗宫颈上皮内瘤样变（CIN）和原位癌。方法 应用TCT液基细胞学薄片检测法对宫颈病变做阴道细胞学分析，对CIN进行分级（Ⅰ、Ⅱ、Ⅲ）鉴定。对高危病人（如ASCUS、CIN、宫颈原位癌）应用基因杂交捕获法（Hybrid Capture，HC-Ⅱ）分型检测HPV，同时用PCR法检测HPV—DNA，进一步做HPV分型鉴定。最后所有病人均进行阴道镜检查及病理组织学诊断。结果 2003年2月～2005年3月将标本送往广州金域医学检验中心进行TCT和HPV检测（HC—H），其中TCT检测1086例，发现不典型鳞状细胞39例、CINⅠ25例、CINⅡ3例、CINⅢ5例，原位癌3例。高危病人（如ASCUS、CIN、宫颈原位癌）进行HC—Ⅱ检测40例，阳性25例（其中原位癌、CINⅢ和CINⅡ均阳性）。结论 HPV感染与宫颈病变，特别是宫颈癌、宫颈癌前病变的发生有明显的相关性，病变越重，HPV的感染率越高。提示宫颈癌的防治重点应放在高危型HPV感染者。     

不同人群自取样和医取样HPV检测用于子宫颈癌筛查分析

目的 比较不同人群阴道自取样和医取样人乳头瘤病毒（HPV）分型检测的效果,探讨自取样HPV分型检测用于子宫颈癌初筛的可行性。方法 2021年4月至2022年9月对江西省靖安县农村地区普通人群女性（n=210）和北京大学人民医院就诊有高危因素女性（n=146）共356例进行阴道自取样和医取样子宫颈癌筛查,采用PCR-荧光探针法进行15种HPV分型检测;对HPV16/18阳性或细胞学异常者（≥ASC-US）进行阴道镜检查及活检。比较不同人群和不同取样方式HPV阳性及相应亚型的符合情况。结果 普通人群女性HPV阳性率为14.29%(30/210),有高危因素女性HPV阳性率为61.64%(90/146)。自取样与医取样HPV检测结果阳性一致347例,不一致9例,符合率为97.47%(347/356)。两种取样方法 HPV检测的阳性率比较,差异无统计学意义（P <0.05）。普通人群HPV亚型分布依次是HPV52、HPV53、HPV58、HPV33和HPV31型;有高危因素女性依次是HPV16、HPV58、HPV51、HPV52和HPV53型,其中两者的医取样和自取样分布基本一致。普通人...     

用DNA杂交法检测HPV的不同型别

本文报告澳大利亚悉尼市患者的宫颈刮屑、肛门刮屑和肛门活检中人乳头瘤病毒(HPV)的不同型别的发病率。所用的方法是将许多份细胞样品点在同一张尼龙膜上,然后相继用HPV DNA6+11型和16+18型两种混合探针与之进行杂交。同时还用另一种探针(Alu探针)估测样品中DNA量是否足够。结果见表一和表二。另外还对7名男性(6名为同性恋者,且有生殖器疣史)的Alu阳性肛门刮屑进行HPV DNA杂交检测。结果为6名同性恋者中,4名既往肛门湿     

表达谱基因芯片筛选子宫颈癌甲基化差异基因的研究

目的筛选人乳头瘤病毒（HPV）阳性宫颈癌细胞系中由于HPV感染诱导沉默的特异抑癌基因,探讨HPV感染可能导致抑癌基因发生甲基化的机制。方法选取HPV阳性HeLa和Caski宫颈癌细胞系和HPV阴性C-33A和HT-3宫颈癌细胞系,分别经去甲基化药物5-氮杂-2′-脱氧胞苷（5-Aza～CdR,Aza）处理,采用Agilent人类全基因组表达谱芯片检测Aza处理前后细胞全基因组的表达水平,采用实时荧光定量PCR（RTqPCR）验证芯片结果,亚硫酸氢盐-基因组测序法（BGS）检测目的基因甲基化水平。结果①芯片结果显示：HPV阳性和阴性细胞系共721条基因表达存在差异。②用药后表达显著上调的基因：HeLa细胞系825条,Caski细胞系815条,c_33A细胞系1023条,HT-3细胞系1196条。HeLa和Caski细胞系共表达上调的基因为182条,剔除阴性细胞系共表达上调基因,筛选出阳性细胞系HPV相关表达上调基因97条,与阴性细胞系基因表达比较,差异有统计学意义（P〈0．01）,最终筛选出13条差异表达基因,其中具有功能者7条。③RT-qPCR结果：筛选基因在宫颈癌细胞系中的表达与芯片检测结果一致;HPV阳性宫颈癌细胞系中甲基化频率明显高于阴性细胞系,支持芯片结果。结论筛选出的7条新的潜在抑癌基因可能由于HPV感染诱导发生甲基化而沉默,为进一步探讨HPV诱导抑癌基因发生甲基化的机制提供实验基础。     

HPV分型与整合检测在子宫颈病变中的诊断价值研究

目的:探讨人乳头瘤病毒(HPV)分型与整合检测对子宫颈癌及癌前病变患者的诊断价值。方法:对2021年1月至2022年8月在苏州大学附属第一医院行HPV分型与整合检测的202例患者进行回顾性研究，以同期行阴道镜下子宫颈组织活检的病理结果为诊断金标准，分析HPV整合检测在子宫颈病变中的诊断价值。结果:Logistic回归分析结果显示年龄、HPV整合结果是患者子宫颈病变程度的影响因素，差异有统计学意义(P<0.05);进一步联合患者HPV分型整合检测结果和年龄建立子宫颈病变的诊断预测模型，其ROC曲线下面积(AUC)为0.915,灵敏度为82.60%,特异度为91.60%。结论:HPV整合状态与子宫颈病变息息相关，HPV分型与整合检测对子宫颈高级别病变的诊断具有显著临床价值，能对HPV阳性患者进行精准分流，可作为阴道镜检测的有效补充，共同评估子宫颈癌及癌前病变进展风险。     

HPV oligonucleotide microarray-based detection of HPV genotypes in cervical neoplastic lesions

BACKGROUND: In this study we examined the use of a new-human papillomavirus (HPV) detection method, the HPV oligonucleotide microarray system (Biomedlab Co., Korea), which we compared with the well-established HPV DNA detection system (Hybrid Capture II; HC-II, Digene Co.). This new method prompted us to develop a new HPV genotyping technique, using the oligonucleotide microarray, to detect the generic and type-specific sequence of HPV types. In particular, we undertook the evaluation of the clinical efficacy of the HPV oligonucleotide microarray for detecting HPV in cervical neoplastic lesions. METHODS: One hundred forty patients were involved and classified into three groups according to their histopathologic diagnoses: Group I (nonspecific chronic cervicitis; n = 61), Group II (low-grade squamous intraepithelial lesion (SIL); koilocytosis, and mild dysplasia; n = 39), and Group III (high-grade SIL; moderate, severe dysplasia and in situ carcinoma; n = 40). Cytological diagnoses were based on the Bethesda System and cervical samples were analyzed by the two methods. The HPV oligonucleotide microarray detected 15 types of high-risk HPV (HPV-16/-18/-31/-33/-35/-39/-45/-51/-52/-56/-58/-59/-66/-68/-69) and 7 types of low-risk HPV (HPV-6/-11/-34/-40/-42/-43/-44). RESULTS: In 105 of the 140 cervical samples (75%), HPV DNAs were examined using the HC-II method. HPV detection rates using the HPV microarray agreed with those of HC-II. One HC-II-positive, but HPV microarray-negative, case occurred in the low-grade SIL (Group II) and was later confirmed negative for HPV. The other HPV microarray-positive but HC-II-negative case was found to be HPV-18 by PCR. Low-risk types of HPV were detected in 3 of 39 low-grade SIL cases (Group II) using the HPV microarray. HPV-16 was the most frequent type (32.1%) in all specimens tested, and was significantly more frequent in low-grade or high-grade intraepithelial lesions (Groups II or III) than in normal controls (Group I) (P < 0.05). HPV-58 was the second most common type (17.5%) in Group III. The HPV microarray was found to have advantages in terms of identifying the HPV genotypes and cases of multiple HPV infection. Double HPV infections were detected in 12 cases and triple HPV infections in 7 cases. Two cases were positive for four types of HPV (HPV-16/18/33/35, HPV-16/18/58/68). The sensitivity of HPV testing (HC-II; 94.9%, HPV microarray; 93.7%) for identifying patients with squamous intraepithelial lesion was significantly better than the sensitivity of cytology (77.1%, P < 0.05). On using multiple logistic regression analysis to estimate the relative risk of SIL versus HPV type, HPV-16-positive cases were found to have a 7.5-fold risk of SIL (95% CI = 3.28-16.51; P < 0.01). HPV-33 and HPV-58 were found to be significantly related to high-grade SILs (P < 0.01). CONCLUSIONS: Our results suggest that the HPV oligonucleotide microarray is highly comparable to HC-II for detecting HPV in cervical specimens. The HPV oligonucleotide microarray provides useful information on viral genotype and multiple HPV infections in HPV-related cervical lesions. Genetic information on HPV in cervical specimens might be a particular benefit of the new procedure in the management of cervical neoplastic lesions     

Detection and typing of HPV genotypes in various cervical lesions by HPV oligonucleotide microarray

OBJECTIVES: This study was conducted to evaluate a clinical efficacy of human papillomavirus (HPV) oligonucleotide microarray (Biomedlab Co., Seoul, South Korea) for the detection of HPVs in various cervical lesions. RESULTS: HPV DNAs from 234 patients were analysed by two methods. Among those, 212 patients were classified into 5 groups according to the histologic diagnosis: chronic cervicitis, cervical intraepithelial neoplasia (CIN) I, CIN II, CIN III, and invasive cervical carcinoma. PCR-RFLP could detect 7 types of high-risk HPVs (HPV 16/18/31/33/35/52/58) and HPV microarray could detect 15 types of high-risk HPVs (HPV 16/18/31/33/35/39/45/51/52/56/58/59/66/68/69) and 7 types of low-risk HPVs (HPV 6/11/34/40/42/43/44).HPV genotyping by HPV oligonucleotide microarray revealed that HPV16 was the most frequent type (30.2%) in all specimens tested and was significantly more frequent in CIN III and invasive carcinomas than other lesions. METHODS: HPV DNAs were detected in 158 and 174 of the 234 cervical samples by PCR-RFLP and HPV microarray, respectively. The correlation between the two methods was good in detecting HPVs in general (kappa index = 0.69) and HPVs 31 and 52 (kappa index = 0.70 and 0.70, respectively) and excellent in detecting HPVs 16, 18, 33, 35, and 58 (kappa index = 0.90, 0.88, 0.92, 0.77, and 0.84, respectively). Double HPV infection was detected in 10 cases and one triple infection was detected. By combining cytology and HPV testing, the sensitivity was improved to 87.5, 95.5, 96.1, and 97.2% in CIN I, CIN II, CIN III, and carcinoma, respectively. CONCLUSIONS: This results suggest that HPV oligonucleotide microarray is a highly comparable method to the previously used PCR-RFLP method for the detection of HPVs in cervical specimens. Genetic informations for HPV infection in cervical specimens may offer new strategies in manipulating the patients harboring cervical intraepithelial neoplasia and cervical carcinoma.     

Genotyping of 22 human papillomavirus types by DNA chip in Korean women: comparison with cytologic diagnosis

OBJECTIVE: More sensitive and reliable methods than individual testing (such as polymerase chain reaction, restriction fragment length polymorphism, and Southern blot) should be developed as screening tools for the detection of latent human papillomavirus. Today, the new Bethesda system recommends human papillomavirus testing as an adjuvant to the conventional Papanicolaou smear for more comprehensive identification of women at certain risk of cervical neoplasia. We performed human papillomavirus genotyping with the newly designed human papillomavirus DNA chip, which is based on polymerase chain reaction for high-throughput screening power, and compared the results with the results of a Papanicolaou smear according to the new Bethesda system. STUDY DESIGN: Polymerase chain reaction amplifications of the human papillomavirus L1 region from biologic samples were hybridized to silanized glass slides by a microarrayer, which comprised 22 specific oligonucleotide probes to their genotypes, consisting of 15 high-risk and 7 low-risk types. Two cervical cancer cell lines and 20 plasmids that contained each type of the human papillomavirus whole genome were used for the evaluation of this method; in all cases, the cancer cell lines and plasmids showed clear positive signals on their corresponding positions. A comparative study that used 685 cervicovaginal swabs was performed by human papillomavirus DNA chip microarray together with Papanicolaou diagnosis. RESULTS: Human papillomavirus was identified as positive in 31.9% of the 414 control samples and in 78.6% of the 271 neoplastic lesions. The major prevailing human papillomavirus genotypes were human papillomavirus types 16, 58, and 18, in descending order of incidence (average overall, 78.8%). Almost all of the remaining cases were comprised of human papillomavirus types 39, 52, 56, and 51. The frequency of multiple infection of human papillomavirus was highest in low-grade squamous intraepithelial lesion but was lowest in squamous cell carcinoma. All cases that exhibited infection of single human papillomavirus type 58 were squamous cell carcinoma. CONCLUSION: Human papillomavirus types 16, 18, and 58 were confirmed to be major causative factors for cervical carcinogenesis. Low-grade squamous intraepithelial lesion is a heterogeneous entity that is composed of different human papillomavirus subtypes and prevails in younger women (<40 years old). The human papillomavirus chip has potential use as a high-throughput screening test.     

HPV detection and genotyping as an earlier approach in cervical cancer screening of the female genital tract

Human papillomavirus (HPV) infection is the leading risk factor for cervical intraepithelial neoplasia (CIN) and cervical cancer. More than 100 virus genotypes have been identified so far, some of them strongly associated with the development of neoplasia. The aim of this study was to evaluate the prevalence of the different HPV genotypes in women presenting no cytological alterations in cervical cells, in women presenting light alterations, and in women presenting severe alterations at routine gynecological examination. We retrospectively analyzed 97 HPV results of women submitted to cervical cancer screening compared to their Papanicolaou and colposcopy examinations. Data were analyzed individually and within groups to correlate the HPV genotypes identified by polymerase chain reaction (PCR) and the respective alterations in cervical cells. Among the nine cases diagnosed as CIN I (9.3%), two were positive for low-risk HPV genotypes (22%), and the other seven were negative for HPV by PCR (78%). CIN II or CIN III diagnoses were associated with positive HPV results by PCR in four cases (36%), for high-risk as well as low-risk genotypes. There were two patients with severe cytological alterations in cervical cells, but with an indeterminate HPV genotype (18%), and one case with a negative HPV result (9%). Among the 57 cases without cytological alterations, seven were positive for low-risk HPV (12%) and two for high-risk HPV genotypes (3.5%). In the 48 remaining cases, we observed one with an indeterminate HPV genotype (2%), and the other 47 were negative for HPV by PCR (47%). Our study demonstrates an important prevalence of high- and low-risk HPV genotypes in our population, including those not present in the commercially available vaccine, even in patients with no evidence of cytological alterations in cervical cells. These results highlight the usefulness of HPV detection and typing as an early approach for cervical cancer screening and prevention.     

Localized distribution of human papillomavirus genotypes in the uterine cervix

INTRODUCTION: The localization and distribution of single or multiple HPV genotypes in the uterine cervix has not been studied thus far. The present study was undertaken to determine whether single or multiple HPV genotypes detected in cervical smears originate from a single (dysplastic) area, or from different areas (dysplastic or normal) of the uterine cervix. METHODS: Of eight patients with moderate or severe dysplasia, 31 colposcopically guided biopsies of different dysplastic lesions of the uterine cervix, as well as of normal epithelium were investigated. A highly sensitive, broad spectrum, short fragment polymerase chain reaction (SPF-10 PCR) HPV detection method in combination with a line probe assay (LiPA) for simultaneous genotyping was used. RESULTS: In the uterine cervix of four of the eight patients, multiple HPV genotypes were detected. These multiple HPV genotypes were detected in different biopsies as well as within a single biopsy. In three patients, all with carcinoma in situ or microinvasive carcinoma, only a single HPV genotype, HPV 16, was found all over the cervix including in the normal epithelium. CONCLUSION: Different HPV genotypes can be detected in different dysplastic lesions as well as within single lesions, especially in patients with severe dysplasia. The severity of the lesion may possibly have a relation with the distribution of the HPV genotypes. The low number of patients and biopsies does not allow definite conclusions. However, the impact of these findings on the outcome of screening and vaccination programs remains to be elucidated.     

Genotyping human papillomaviruses: Development and evaluation of a comprehensive DNA microarray

GoalsTo define the analytical and clinical performance of a human papillomavirus (HPV) custom-designed microarray targeting the HPV L1 gene for viral genotyping.MethodsMicroarray probes were designed by cataloging the genome sequence of all 120 known HPV types to generate tiling probes using eArray® software against the unique L1 capsid gene segments targeted by MY09/11 and FAP59/64 primers. The microarray (1 slide × 8 arrays × 60 K features) synthesized in situ by inkjet printing was tested using synthetic type-specific HPV DNA and existing HPV DNA from cervical cytology. The synthetic HPV L1 segments (genotypes 6, 11, 16, 18, 31, 33, 35, 45, 53, 58, 66, 73, 83) were manufactured from sequences stored in the NCBI taxonomy database. Using the hybridization patterns of the synthetic HPV DNA as the Support Vector Machine classifier, HPV DNA from patient samples were genotyped and compared to antecedent DNA sequencing/BLAST® results for concordance.Results16 cytology-derived HPV DNA samples and 13 synthetic type-specific HPV DNA samples were tested singly, in duplicate, or in combination on 40 arrays. The synthetic HPV DNA hybridization patterns were found to be uniquely distinctive to serve well as a classifier of unknown HPV-containing specimens. For the 16 HPV DNA + samples classified, 15 were concordant with DNA sequencing results. In 6/16 (38%) samples, the microarray hybridization pattern revealed ≥ 2 concurrent HPV infections.ConclusionThe novel “HPV Array” was sensitive and specific for detecting single and multiple infections. This proof-of-principle project demonstrated the accuracy and advantages of microarray technology for HPV genotyping.     

Development and evaluation of a highly sensitive human papillomavirus genotyping DNA chip

OBJECTIVES: Test of human papillomavirus (HPV) is a useful adjunctive tool of Pap smear to screen cervical cancer. We have developed a novel HPV genotyping DNA chip arrayed by multiple oligonucleotide probes of both L1 and E6/E7 gene sequence of 42 types of anogenital HPV. METHODS: Consensus PCR products of L1 and E6/E7 gene sequences of HPV are hybridized to arrayed probes on the HPV chip and HPV genotypes are identified by fluorescence scanner. We have comparatively analyzed the value of HPV DNA chip and DNA sequencing in 100 cervical cancer tissues. RESULTS: Overall, 98 cervical cancer tissues were found to harbor DNA sequences of high-risk type HPVs, of which 88 (89.8%) were detected by PCR-sequencing of L1 alone, 98 (100%) by PCR-sequencing of both L1 and E6/E7, and 98 (100%) by HPV DNA chip, respectively. All of the genotypes of HPV detected on sequencing analysis were also found on DNA chip analysis. HPV DNA chip was superior to direct DNA sequencing in detection of mixed infection. CONCLUSIONS: These results suggest that HPV DNA chip analysis in the present study is highly accurate for detection and genotyping of HPV and may have potential value as a robust, high-throughput screening test of uterine cervix cancer.     

海南地区宫颈癌的HPV基因型检测及其临床意义

目的:通过对宫颈癌组织的HPV分型检测,了解本地区宫颈癌HPV感染率及其亚型的分布情况,并分析HPV感染与宫颈癌临床生物学行为之间的关系.方法:收集200例宫颈癌患者病灶组织标本,采用HPV分型基因芯片检测系统,同时对23种HPV亚型(18种高危型和5种低危型)进行检测.结果:①HPV阳性率94.00%,共检出HPV16(69.68%)、HPV 18(16.49%)、HPV 33(1.06%)、HPV 45(1.60%)、HPV 58(1.06%)、HPV、73(0.53%)6种高危型亚型.其中HPV 16+18(7.45%)、HPV 16+45(1.06%)、HPV 18+33(1.06%);②鳞状细胞癌的HPV检出率明显高于腺癌(P<0.05);③HPV阳性率与宫颈癌患者的年龄、临床分期、病理组织学分级、盆腔淋巴结转移、治疗后的复发差异均无统计学意义(P>0.05).结论:本组宫颈癌的HPV感染率很高,且均为高危型,其中以16型和18型占绝对优势;HPV亚型比较局限,仅有6种;HPV多重感染并不多见;高危HPV的存在与鳞状上皮恶变关系密切.     

黑龙江省铁力市育龄妇女宫颈癌前病变筛查的研究

目的:检测目标人群中宫颈病变的现状及人乳头瘤病毒(human papillomavirus,HPV)流行病学特征。方法:(1)应用醋酸白试验(VIA)、碘不着色试验(VILI)对4007例妇女宫颈行肉眼观察,疑似异常病例行阴道镜检查,异常病例组织活检确诊;(2)采用导流杂交基因分型技术(HybriMax),对宫颈癌及CIN(cervical intraepithelial neopla-sia)患者进行21种HPV基因型的分型检测,分析HPV感染率以及宫颈疾病发病状况。结果:(1)两年诊断出宫颈病变发病率为5.01%,其中CINⅠ3.74%,CINⅡ0.87%,CINⅢ/原位癌0.32%,浸润癌0.07%;(2)病变宫颈HPV-DNA基因分型检测到HPV-DNA阳性127例,阳性率63.18%,其中CINⅠ53.33%,CINⅡ91.43%,CINⅢ92.30%,宫颈浸润癌100%;高危型HPV感染136例次,占77.27%,低危型40例次,占22.73%;单一亚型感染89例,占70.08%,多重HPV基因型混合感染38例,占29.92%。感染最多的基因型是HPV16,共45例次,占25.57%,其次是HPV58,占10.23%,随后基因型由高到低依次为:HPV52,占10.23%;HPV33,占7.95%;HPV6,占6.82%,不同级别CIN及浸润癌HPV感染率差异有统计学意义(P0.05)。结论:(1)肉眼观察筛查宫颈病变虽有一定的漏诊率,但由于检查成本低廉,在经济欠发达地区,仍可作为防癌普查的基本方法;(2)HPV16型是本地区宫颈病变HPV感染的主要型别,位居第二为HPV58,HPV52、HPV33、HPV6分别位居第三、四、五位。加大对高危人群的普查力度势在必行。     

Good correlation of HPV DNA test between self-collected vaginal and clinician-collected cervical samples by the oligonucleotide microarray

OBJECTIVES: To evaluate the efficacy of self-collected vaginal samples for high-risk HPV detection by the HPV oligonucleotide microarray method (HPVDNAChip). METHODS: One hundred and eighteen patients with abnormal Pap smears were included. Self-collected vaginal and clinician-collected cervical samples for HPV testing were obtained. The result of the HPV DNA test was compared with the histopathological diagnosis or colposcopic finding. RESULTS: Of the 118 patients, 42 (35.6%) had >or= cervical intraepithelial neoplasia (CIN) III lesions. Using the HPVDNAChip, high-risk types of HPV were detected in 38 of these 42 patients (90.5%) with the self-collected vaginal samples and in 37 of 42 (88.1%) with the clinician-collected cervical samples. The agreement of HPVDNAchip results between self- and clinician-collected samples was very good (kappa = 0.81) with a 93.2% concordance rate. Multiple HPV infections were found in 17 of 88 (19.3%) HPV-positive clinician-collected cervical samples. The rate of multiple HPV infection tended to decrease as the degree of pathologic classification increased. CONCLUSION: Using the HPVDNAchip to assay for HPV infection, results from self-collected vaginal samples were compatible with those from clinician-collected cervical samples.