1516例性病门诊患者人乳头瘤病毒感染情况及其基因型别

目的了解性病门诊患者人乳头瘤病毒(HPV)的感染情况及其基因型别。方法用导流杂交分型检测法对1 516例性病门诊患者进行HPV检测与基因分型,评价HPV及其基因型别与疾病类型的关系。结果 1 516例受检者中,HPV总阳性率为55.61%。基因型分析发现,26例鲍温病样丘疹病患者均有高危型HPV感染,而762例尖锐湿疣患者中低危型HPV感染占70.86%。21种HPV基因型均被检出,以HPV6(28.86%)、11(17.23%)、16(10.41%)型感染最为常见。检出的843例阳性患者中单一型别感染占61.68%,以HPV6和HPV11感染为主;HPV多重感染以高低危型HPV混合感染为主。结论 HPV基因型别分布与疾病类型密切相关,HPV亚型的鉴别对相关疾病的预防和治疗具有临床应用价值。     

MALDI-TOF-MS技术检测宫颈癌中HPV感染状态

目的:本研究拟评MALDI-TOF-MS技术在HPV分型中的实际应用效果.方法:采用MALDI-TOF-MS技术检测河南安阳地区94例宫颈癌样本中HPV感染状态,并与普通PCR方法得到的结果相比较.结果:94例宫颈癌组织中HPV感染率为92.6%,共检测到10种常见的高危型别,以16为主,其次是58和18型.样本单一HPV型别感染为主,其中16型单一型别感染占6I.7%,见多种型别和组合的多重感染.与普通PCR检测结果比较发现两种方法样本HPV(不分型)检测一致率为95.7,样本HPV16型检测一致率为93.6%.结论:MALDI-TOF-MS技术能够精确、快速、高通量的对宫颈癌中HPV进行检测和分型.     

尖锐湿疣病变的人乳头瘤病毒6型L1序列多态性分析

采用PCR方法从协和医院尖锐湿疣患者皮损中检测人乳头瘤病毒(HPV),并通过限制性片断长度多态性分析分型发现,HPV6型为主要感染型别,其次是HPV11型.根据临床特征选择主要致病型HPV6 8个分离株,扩增L1晚期基因,构建重组测序质粒,双脱氧法测序并分析L1区的核苷酸和氨基酸序列变异状况.结果表明,HPV6L1基因序列发生碱基替换的区域主要有四个区,包括SR1(5911～6104),SR2(6217～6273),SR3(6540～6661)和SR4(7062～7250).少数的碱基替换导致错义突变,推导的蛋白质一级结构中有0～3个氨基酸发生变异,但抗原性强弱与原型HPV6基本一致.     

高通量流式荧光微球悬液芯片检测乙肝病毒基因分型

目的:建立HBV基因分型高通量液相芯片检测技术,并探讨其应用价值.方法:对GenBank中收录的明确分型的HBV基因序列进行分析,选择preS2-S区设计引物和A、B、C和D型特异性探针.与荧光编码微球偶联的特异型探针与一条引物生物素标记的PCR产物直接杂交反应,然后结合亲和素标记的藻红蛋白,用流式检测仪(Bio-Plex 200)检测荧光信号.检测182份阳性乙肝患者血清DNA,其中35份样品检测结果与测序法比较.用B、C型质粒DNA倍比稀释及混合样品检测灵敏度来评估该方法.结果:建立了HBV基因分型的快速高通量液相芯片检测方法.182份患者血清检测结果为:B型占24.2％ (44/182),C型占71.4％(130/182),D型为6.6 ％(12/182),BC混合型4.4％(8/182).其中35份样本与测序法比较,除3份混合型测序法未检出外,其它32例结果均相同本方法的灵敏度检测下线为1×103 copies/mL.结论:应用悬液芯片技术进行乙肝病毒的基因分型分析,具有较好的特异性和较高的灵敏度,并有简便、灵活和高通量等优势.该检测系统不仅在科研中有广泛的前景,也有望成为临床推广的多重分子诊断和基因分型的新方法.     

HPV分型及其高危亚型与宫颈病变的关系

目的研究HPV分型及其高危亚型与宫颈病变程度的关系。方法采用HPV基因分型技术对2014年7月至2015年8月在广州中医药大学第一附属医院妇科门诊就诊和体检的3 142例患者进行HPV分型检测,其中327例患者进行宫颈病理检查,分析高危型HPV感染与宫颈病变程度的关系。结果 3 142例受检对象中,21种HPV型别均被检测出,阳性检出者共969例,总检出率为30.80%,其中高危型741例,检出率为23.58%。随着宫颈病变级别的升高,高危型HPV阳性率也相应升高(P0.05),HPV16型感染阳性者宫颈癌发生率最高(P0.05)。高危型HPV阳性者中单一感染和多重感染与宫颈病变程度无关(P0.05)。结论高危型HPV亚型检测在宫颈病变预防和诊治中具有重要意义。     

尖锐湿疣样本中HPV病毒的分子检测

调查男性和女性尖锐湿疣样本中人乳头瘤病毒（HPV）的检出率及病毒类型,为研发相关防治疫苗提供依据,以HPV 外壳蛋白DNA序列为模板设计特异引物,SSP－PCR扩增检测样本中HPV的感染率和病毒类型。收集了北京及邯郸市医院门诊尖锐湿疣样本22例,其中男性13例,女性9例。检测发现所有样本中存在着高浓度的HPV病毒DNA。男性样本中有5例感染HPV6型,6例感染HPV11型,2例为HPV6＋HPV11混合感染。女性样本中有3例感染HPV6型,2例感染HPV11型,4例为 HPV6＋HPV11混合感染。被诊断为宫颈湿疣的4位女性还在其含宫颈粘膜脱落细胞的样本中检出了HPV16、HPV18、HPV33、HPV35、HPV45、HPV54、HPV56或HPV58等高危险型病毒类型。所有检测到的HPV病毒DNA片段均TA克隆并将测定的DNA序列存入了国际基因数据库GenBank（DQ003066－DQ003079）。调查没有在单纯的男女尖锐湿疣组织块中检测到除HPV6和HPV11以外的其他HPV类型。该研究建立了灵敏可靠的HPV分子检测及分型方法,尖锐湿疣中HPV的检出率达100％。 本研究初步结果显示导致男女尖锐湿疣的HPV病毒类型没有显著差异,主要为HPV6及HPV11型。     

宫颈癌患者人乳头瘤病毒(HPV)主要型别及其感染研究

本文探讨了江西省和广东省宫颈癌患者人乳头瘤病毒(Human papillomavirus,HPV)感染及其型别分布,分析了高危型HPV对各种宫颈病变的感染情况,为宫颈癌的早期发现和临床诊治提供科学依据。首先采用细胞学、HPV DNA检测(第二代杂交捕获法,HC2)、电子阴道镜和宫颈化学着色方法筛查宫颈癌患者,经病理镜检确诊,然后用GP PCR-SBT法对宫颈癌患者进行HPV基因分型。江西省溪口镇、古市镇及修水县城宫颈癌癌前病变发生率为5．7‰。HC2方法发现宫颈癌患者13种高危型HPV DNA阳性率为89．9％,宫颈上皮内瘤样病变的为84．8％,对照组为24．5％。采用GP PCR-SBT方法进行基因分型发现,江西省宫颈癌患者存在HPV16、58、31、33、18、66、6、11、56和81十种型别,其中HPV81型在国内外鲜有报道。据此提出生殖道高危型HPV感染是妇女宫颈癌发病的重要因素。并发现江西省宫颈癌高发区妇女高危型HPV感染率为24．5％。建立了HPV基因分型的方法,对HPV致宫颈病变的分子机制进行了分析。     

HPV基因分型检测在宫颈癌筛查中的应用

目的:探讨人乳头瘤病毒(human papilloma virus,HPV)基因分型技术在宫颈癌筛查中的临床应用价值。方法:采用反向斑点杂交法对HPV基因型进行分析,包括低危型(如6、11、42、43、44型等)和高危型(如16、18、31、33、35、MM4型等)共23个型别。结果:98例样本中检出HPV阳性者38例,23种基因型中共检出15种HPV型别,HPV总感染率38.8%(38/98)。慢性宫颈炎患者中HPV感染率30.4%(14/46),检测到的HPV亚型主要是HPV43,11,6,42,31和51(其中HPV6,42,31,51检出率一致)。CINI患者HPV感染率15.4%(4/26),最常见的HPV亚型为HPV35,其次为HPV16,52,53和59(其中16,52,53,59检出率一致)。CINⅡ患者HPV感染率71.4%(10/14),以HPV16,6,11,51等亚型最常见。CINⅢ患者HPV感染率75%(6/8),HPV亚型以HPV16,18,31,53为主。SCC患者HPV阳性率为100%(4/4),HPV16检测率最高,其次为HPV52,18,33,58,59和66。结论:持续的HPV感染与宫颈疾病有着密切的关系。HPV基因型的检测为临床进行宫颈疾病的筛查、诊断、治疗以及预后观察提供了较大的帮助。     

应用树状DNA杂交(DDH)对生殖道尖锐湿疣中HPV DNA的分型检测

从手术切除的50例生殖道尖锐湿疣新鲜标本中,以及15例正常人血清中,提取基因组DNA,同时用树状DNA杂交(dendrimer DNA hybridizalion,DDH)技术和PCR进行HPV DNA的分型检测.结果50例尖锐湿疣中,以DDH方法检测,感染HPV6型者20例,感染11型者24例,6/11型混合感染者3例,阴性3例,总检测率达94%;以PCR方法检测,HPV6型感染者21例,11型感染者24例,6/11型混合感染者3例,阴性2例,总检测率为96%.15例正常人血清中,以DDH方法检测,HPV感染的假阳性率为0%;以PCR检测,假阳性率为6.67%.还以HPV阳性标本对DDH方法做了敏感度的测定,结果阳性病例DNA检测最低浓度为97.28pg/ml.研究表明,DDH技术具有较高敏感性和高特异性,且成本较低,操作安全简便,可适用于基层中小医院较大样本量筛查.     

寡核苷酸芯片法、实时PCR 和测序法进行乙肝病毒 基因分型的比较

目的:比较寡核苷酸芯片法、实时荧光PCR和测序法在对慢性乙肝患者病毒基因分型的比较和方法学评价。方法:对126例不同基因型的慢性乙肝患者的血清样本分别用寡核苷酸芯片法、实时荧光PCR法和测序法进行基因分型,并评价各种方法的临床表现、所需时间和检测成本。结果:寡核苷酸芯片法、实时荧光PCR分别能检测到1%和0.1%比例的基因型。在126例慢性乙肝患者的临床样本中,寡核苷酸芯片法、实时荧光PCR和测序法分别检测出41(33%)、41(33%)和45(36%)例为B型,76(60%)、76(60%)、81(64%)例为C型。寡核苷酸芯片法、实时荧光PCR均检出9例B、C混合基因型。在三种检测方法中实时荧光PCR是最快速和廉价的。结论:寡核苷酸芯片法、实时荧光PCR能检出B、C混合基因型,而测序法只能检测出样本的主导基因型。     

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer.     

The evaluation of human papillomavirus genotyping in cervical liquid-based cytology specimens; using the Roche Linear Array HPV genotyping assay

Objective:  To ascertain the usefulness of the Roche Linear Array human papillomavirus (HPV) genotyping assay for assessing HPV genotypes in liquid-based cytology (LBC) samples and to evaluate this methodology within a cytopathology laboratory. These tests are of importance as persistent infection with high-risk HPV genotypes is considered a causal factor in the development of cervical cancer.
Methods:  A total of 175 cervical LBC samples were tested using the Roche Linear Array HPV genotyping test. The suitability of the assay use in routine cytopathology laboratory was considered. HPV genotypes were matched to the cervical cytology results, which included negative, borderline nuclear abnormalities, mild, moderate and severe dyskaryosis.
Results:  The assay could be applied to screening samples with the combined result available at the reporting stage. There were no test failures. All samples used after cytological analysis had sufficient DNA for testing. The results were reproducible and easily read and there was concordance of results between biomedical scientists. The results of the assay showed co-infection with multiple HPV genotypes was common in both high-grade and low-grade cytology samples. The percentage of HPV+ samples in the normal cytology samples (although in this grouping the number of samples was low) was 37%. In the cytology samples reported as severe dyskaryosis the HPV genotypes most commonly found were HPV16 and HPV51.
Conclusion:  The assay was able to detect multiple HPV infection with a wide range of genotypes in LBC samples sent for routine cytological analysis. It would be suitable for use in a cytopathology laboratory. The results of the assay show that the genotype profile has some variation from other geographical regions, and more work is needed to determine population prevalence, to ascertain the impact of the HPV vaccine, to evaluate test for cure and HPV triage management.     

Prevalence and correlation of human papillomavirus genotypes with clinical factors in cervical samples from Mexican women

In the last decade, the inclusion of HPV DNA testing in cervical cancer screening has provided one of the best strategies for the prevention and timely detection of HPV. We conducted a high-throughput HPV genotyping study based on MALDI-TOF mass spectrometry to determine the prevalence of 24 HPV genotypes, including oncogenic genotypes, in Mexican women and correlated the results with cytological findings and clinical variables. We likewise identified the risk factors in patients with the HPV infection. Our study included 1000 women from Sonora, Mexico, who participated in cervical cancer screening campaigns and who underwent a Pap smear and HPV DNA test. The results showed that the overall prevalence of HPV was 27.2%, 18.5% with single, and 8.7% multiple infections. The low-risk HPV genotype 6 (8.5%) and oncogenic genotypes 31 (8.1%) and 53 (4.4%) were the most prevalent in the study population. The number of lifetime sexual partners, previous STIs, and age at first intercourse was significantly associated with HPV infection (P ≤ 0.05). Smoking (OR = 1.5609; 95% IC 1.062–2.292) and more than three lifetime sexual partners (OR = 1.609; 95% IC = 1.124–2.303) represented risk factors for HPV infection. Cytological abnormalities were found in 3.4% of the HPV-positive samples. CIN 1–3 occurred in 0.6% of high-risk HPV cases. In general, the prevalence of the HPV genotypes is high in Mexican women with normal cytological findings. This issue highlights the importance of HPV research in seemingly healthy women and could help guide screening strategies for cervical cancer prevention in Mexico.Impact statementWe are submitting data regarding the prevalence and type distribution of the HPV infection and the risk factors associated with it, which may provide a valuable reference to reinforce screening strategies, and to maintain HPV genotype surveillance in Mexico. We discuss the overall prevalence of HPV infection as detected in normal cytological samples stratified by age, different types of infection, and oncogenic capacity. One of the most important findings was that common HPV genotypes detected in healthy women were the genotype numbers: 6, 31, 16, and 56, likewise, smoking and having a history of more than three sexual partners over their lifetime, represented the main risk factors in this study. Furthermore, we found a low frequency of cytological abnormalities and CIN 1–3 in women with HR-HPV.     

Resequencing Microarray Technology for Genotyping Human Papillomavirus in Cervical Smears

There are more than 40 human papillomaviruses (HPVs) belonging to the alpha genus that cause sexually transmitted infections; these infections are among the most frequent and can lead to condylomas and anogenital intra-epithelial neoplasia. At least 18 of these viruses are causative agents of anogenital carcinomas. We evaluated the performance of a resequencing microarray for the detection and genotyping of alpha HPV of clinical significance using cloned HPV DNA. To reduce the number of HPV genotypes tiled on microarray, we used reconstructed ancestral sequences (RASs) as they are more closely related to the various genotypes than the current genotypes are among themselves. The performance of this approach was tested by genotyping with a set of 40 cervical smears already genotyped using the commercial PapilloCheck kit. The results of the two tests were concordant for 70% (28/40) of the samples and compatible for 30% (12/40). Our findings indicate that RASs were able to detect and identify one or several HPV in clinical samples. Associating RASs with homonym sequences improved the genotyping of HPV present in cases of multiple infection. In conclusion, we demonstrate the diagnostic potential of resequencing technology for genotyping of HPV, and illustrate its value both for epidemiological studies and for monitoring the distribution of HPV in the post-vaccination era.     

安康地区人乳头瘤病毒感染特征及其与宫颈癌的关系研究

目的:调查安康地区女性人乳头瘤病毒(HPV)感染的基因型别及年龄分布特征,分析其与宫颈癌的关系,为宫颈癌防治及HPV疫苗研发提供可靠的依据。方法:收集2010年6月-2012年8月间在本院及安康市部分县级医院妇产科就诊的2736名女性的液基细胞学和组织学标本,分为8个年龄组:16-24岁119例、25-29岁230例、30-34岁343例、35-39岁472例、40-44岁574例、45-49岁512例、50-54岁206例、55-86岁280例,进行病理学分类及HPV分型检测,分析不同年龄组及不同类型宫颈组织中的HPV感染率。结果:2736例女性中发生HPV感染720例(26.32%),共检出21种型别,感染率最高的基因型别是HPV16(25.05%),其他常见型别依次为HPV58、HPV52、HPV6、HPV11。单一感染占76.25%,多重感染占23.75%。HPV感染率在16-24岁、35-39岁和55-86岁三个年龄段出现高峰;而高危型HPV的感染率在35-39岁和55-86岁两个年龄段分别出现高峰。HPV的检出率随着宫颈病变的严重程度而增加,其中正常或炎症人群的HPV感染率显著低于宫颈病变及宫颈鳞状细胞癌患者(均P0.05),且意义未明的不典型鳞状细胞(ASCUS)、CIN1-3及宫颈鳞状细胞癌患者的HPV感染率对比结果存在显著差异(P0.05)。CIN1组、CIN2-CIN3组及宫颈鳞状细胞癌组单一感染率逐渐增加(P0.05),且其二重、三重感染率比较差异均有统计学意义(P0.05)。结论:安康地区HPV16型别感染较广,临床需加强对HPV16型单一感染宫颈病变患者的癌症预防工作。     

铜陵市2815例女性高危型HPV感染状况及基因型分布特征

目的了解铜陵市女性高危型人乳头瘤病毒(HPV)感染状况及基因型分布特征,为女性宫颈癌的预防和治疗提供依据。方法收集2016年8月至2018年12月在本院就诊和体检的2815例女性为研究对象,留取宫颈脱落细胞标本,用荧光PCR法对其进行HPV基因分型检测,并分析其分布特征。结果2815例标本中共检出HPV阳性714例,总感染率25.36%;感染率前3位的分别是HPV16、HPV52和HPV58型,以单一亚型感染为主。不同年龄组患者HPV感染率比较差异有统计学意义(χ^2=23.654,P<0.001)。住院、门诊和体检组女性HPV感染率分别为55.00%、26.21%和7.56%,3组间比较差异有统计学意义(χ^2=123.451,P<0.001)。结论铜陵市女性HPV感染率较高,以HPV16、HPV52和HPV58型感染为主,≥56岁人群感染率最高。     

1235例妇科门诊患者高危型HPV感染状况及亚型分布调查研究

目的了解本地区妇科门诊患者宫颈高危型HPV感染状况及亚型分布,为今后的宫颈癌前病变、宫颈癌防治提供临床依据。方法采用基因芯片技术对1 235例妇科门诊患者进行HPV筛查,筛查出的阳性患者应用流式荧光杂交法进行高危型HPV亚型检测,分析比较宫颈炎、宫颈鳞癌及宫颈腺癌患者高危型HPV感染情况及亚型分布差异。结果六安市金安区妇幼保健院妇科门诊患者HPV感染率高达60%,其中高危型HPV感染率为43. 2%,主要以HPV16、HPV18为主;低危型HPV感染率为30.0%,主要以HPV11为主;单一感染阳性率为34. 1%,而混合型感染高达65. 9%,且两者均以HPV16型和HPV18型为主。宫颈炎患者HPV16型、HPV18型及HPV16 ＋ HPV18型的检出率明显低于宫颈鳞癌和宫颈腺癌患者,其中宫颈腺癌患者HPV16 ＋ HPV18型混合感染率最高。结论妇科门诊患者HPV感染率较高,宫颈癌患者HPV16及18型检出率显著高于宫颈炎患者,加强HPV高危基因型的监测有助于预警宫颈癌尤其是宫颈腺癌的发生。     

宫颈癌患者人乳头瘤病毒感染分布情况及多重感染与临床病理特征的关系

目的:研究宫颈癌患者人乳头瘤病毒(HPV)感染的分布情况及多重感染与临床病理特征的关系。方法:选择2015年1月-2018年1月期间我院收治的118例宫颈癌患者,根据患者宫颈癌的病变程度分为I期组(n=21)、II期组(n=46)、III期组(n=49)、IV期组(n=2)。所有患者均进行HPV分型检测,比较不同程度的宫颈癌患者的HPV感染情况,分析不同宫颈癌病变程度患者的多重感染和临床病理特征关系。结果:118例患者中有97例患者感染了HPV,感染率为82.20%,且II期组、III期组、IV期组患者HPV感染率高于I期组(P0.05)。II期组、III期组、IV期组患者一重感染率低于I期组,IV期组二重感染率低于I期组,II期组、III期组、IV期组患者多重感染率高于I期组,且IV期组多重感染率高于II期组、III期组(P0.05)。多重感染患者类型有多种,其中尤以HPV16+18+53型最多,占比49.05%,其次是HPV16+18+68型感染,占比32.07%,HPV16+53+58型感染,占比13.21%。年龄在50岁以上、分期为III-IV期、鳞癌、淋巴结转移的患者HPV多重感染率更高(P0.05)。结论:HPV多重感染与宫颈癌病变程度和临床病理特征均有联系,对年龄较大且HPV多重感染的宫颈癌患者进行筛查,预防病情恶化。