安阳地区不同食管鳞癌标本HPV感染率的比较研究

目的 对安阳地区的食管鳞癌新鲜手术标本和石蜡标本进行HPV检测,比较两种不同标本来源食管鳞癌HPV感染检测结果,统计学分析二者之间的差异,并同其他区域检测结果进行比较,分析不同标本对检测结果的影响.方法 对安阳的23例新鲜手术标本及23例石蜡标本分别进行核酸提取;分别用通用引物PCR鉴定,并用人乳头瘤病毒核酸扩增分型检测试剂盒对样品进行分型检测;比较两种标本来源检测结果并进行统计分析;比较分析检测结果与其他地区检测结果的差异.结果 经检测新鲜标本HPV感染率为82.6％,其中HPV16型感染率34.8％,HPV18型感染率34.8％.石蜡包埋组织标本HPV感染率78.2％,其中HPV16型感染率30.4％,HPV18型感染率17.4％.结论 首次证明两种来源标本对HPV检测结果无显著性影响,均可作为检测HPV的样本,为HPV科研检测提供更广的标本来源.     

应用聚合酶链反应检测食管癌组织中人乳头瘤病毒

目的 探讨人乳头瘤病毒(human papillomavirus,HPV)与我国河南地区食管癌发生的相关性.方法 应用HPV L1通用引物GP5+/6+、HPV16E6和HPV18E6型特异性引物多聚酶链反应(Polymerase chain reaction,PCR),检测林州市食管癌组织中HPV存在状况.结果 31例食管癌组织中,29例检测到HPV阳性,阳性率为93.5%;其中19例检测到HPV16 E6基因,阳性率为61.3%,8例为HPV18 E6基因阳性,阳性率为25.8%,5例HPV16E6和18E6基因均阳性,为混合感染.HPV16和18型阳性率为71.0%.结论 我国河南省林州市食管癌组织中有HPV存在,并且HPV感染可能是食管癌发生的重要病因.     

新疆妇女子宫颈病变组织中HPV16E6基因突变分析

目的通过检测HPV16E6基因突变在新疆维吾尔族和汉族妇女宫颈癌组织中的分布规律,以探讨该突变与宫颈癌发生的关系。方法取汉族宫颈炎(含石蜡包埋组织及宫颈刷液基标本)125例,维吾尔族宫颈炎(含石蜡包埋组织及宫颈刷液基标本)124例;汉族石蜡包埋宫颈癌35例,维吾尔族石蜡包埋宫颈癌共109例。用以上HPV16阳性DNA模板PCR扩增HPV16E6全长基因,PCR产物直接测序,分析新疆女宫颈癌组织HPV16E6基因的突变。结果PCR检测结果显示汉族族宫颈炎组织中HPV16E6阳性率为35.71%(15/42);维吾尔族宫颈炎组织中HPV16E6阳性率为30.46%(14/46);汉族族宫颈癌组织中HPV16E6阳性率为33.33%(2/6),维吾尔族宫颈癌组织中HPV16E6阳性率为22.22%(12/54);宫颈炎与宫颈癌中HPV16E6阳性表达无统计学意义(P>0.05),对21份(上皮内低度病变1例,原位癌1例,低分化宫颈癌4例,中分化宫颈癌4例,高分化1例,轻度炎症4例,中度炎症3例,重度炎症1例,正常宫颈2例)HPV16E6扩增片段的双向测序,其中5例成功测序得到一级结构,并且序列分析表明,2例(维吾尔...     

安阳地区不同食管鳞癌标本HPV感染率的比较研究

目的对安阳地区的食管鳞癌新鲜手术标本和石蜡标本进行HPV检测,比较两种不同标本来源食管鳞癌HPV感染检测结果,统计学分析二者之间的差异,并同其他区域检测结果进行比较,分析不同标本对检测结果的影响。方法对安阳的23例新鲜手术标本及23例石蜡标本分别进行核酸提取;分别用通用引物PCR鉴定,并用人乳头瘤病毒核酸扩增分型检测试剂盒对样品进行分型检测;比较两种标本来源检测结果并进行统计分析;比较分析检测结果与其他地区检测结果的差异。结果经检测新鲜标本HPV感染率为82．6％,其中HPV16型感染率34．8％,HPV18型感染率34．8％。石蜡包埋组织标本HPV感染率78．2％,其中HPV16型感染率30．4％,HPV18型感染率17．4％。结论首次证明两种来源标本对HPV检测结果无显著性影响,均可作为检测HPV的样本,为HPV科研检测提供更广的标本来源。     

新鲜和石蜡包埋宫颈癌组织中HPV l6/18检测

目的为了了解宫颈癌新鲜组织和石蜡包埋宫颈癌组织中HPV检测的差异。方法FQ-PCR技术用于77例宫颈新鲜组织和44例石蜡包埋的宫颈组织中HPVl6/18的检测。结果宫颈癌新鲜组织HPV的检出率为51.43%(36/70),石蜡包埋的櫖颈癌组织检出率为22.72%(10/44)P=0.0024)。两组病毒拷贝对三值分别为5.75±2.36和3.21±1.98(P=-0.000317)。结论宫颈癌新鲜组织HPV的检出率显著高于石蜡包埋的宫颈癌组织,病毒拷贝数有极显著差异。     

保定地区食管癌患者癌组织中人乳头瘤病毒感染的检测

目的 探讨人乳头瘤病毒(human papillomavirus,HPV)与我国河北省保定地区食管癌发生的相关性.方法 应用HPV LI通用引物GP5+,6+、HPV16 E6和HPV18 E6型特异性引物多聚酶链反应(Polymerase chain reaction,PCR),检测保定地区食管癌组织中HPV存在状况.结果 42例食管癌组织中,37例检测到HPV阳性,阳性率为88.1%;其中19例检测到HPV16 E6基因,阳性率为45.2%,8例为HPV18 E6基因阳性,阳性率为19.0%,5例HPV16 E6和18 E6基因均阳性,为混合感染.结论 我国河北省保定地区食管癌组织中有HPV存在,并且HPV感染可能是食管癌发生的重要影响因子.     

应用BIOMED-2引物检测眼附属器淋巴瘤Ig基因重排的初步研究

目的 初步评价BIOMED-2引物在辅助诊断眼附属器淋巴瘤的应用价值.方法 收集63例眼附属器淋巴瘤,均为甲醛固定的石蜡包埋标本,提取基因组DNA并通过扩增管家基因β-actin检测其质量,应用BIOMED-2标准化基因重排检测系统中IgH_B和IgK_B两套多重PCR引物进行Ig基因的PCR扩增,并利用基因扫描技术对扩增产物进行克隆性分析.结果 76.2%(48/63)淋巴瘤石蜡包埋标本的DNA可扩增出300 bp大小的β-actin,适于基因重排检测.IgH_B和IgK_B多重PCR引物的淋巴瘤检出率分别为79.2%(38/48)和68.8%(33/48),二者联合的检出率为91.7%(44/48).结论 应用较少的BIOMED-2引物结合基因扫描技术能检测出大多数眼附属器淋巴瘤,对临床病理诊断具有较高的辅助价值.     

林州市地区食管癌活检组织中人乳头瘤病毒的研究

目的探讨在我国河南省林州市地区食管癌（esophageal carcinoma,EC）活检标本中人乳头瘤病毒（humanpapillomavirus,HPV）,特别是高危型HPV的感染状况。方法收集的食管癌活检标本,使用通用引物的套式PCR反应检测HPV的核酸,分别使用型特异性PCR检测HPV16和18的感染。结果18例活检标本全部为HPV阳性,其中HPV16的阳性率为13／18,HPV18的阳性率为4／18,HPV16／18的复合感染为4／18。结论我国河南省林州市地区食管癌活检组织中有HPV存在,其中HPV16的感染占很大比例,并且HPV感染可能是食管癌发生的重要病因。     

石蜡淋巴瘤组织中IgH FR3区基因重排引物分析

目的 利用生物信息学方法分析免疫球蛋白重链(IgH)框架区(FR3)基因重排引物并探讨其在石蜡包埋非霍奇金淋巴瘤(NHL)组织中应用价值.方法 通过Chustal W 软件比较44条有效的IgH可变区和6条J区的基因片段,选取IgH FR3区3对(P1,P2,P3)基因重排引物,其中,P2为改进的引物.另选一对TCRγ脚引物作为T细胞淋巴瘤重排引物,通过PCR扩增,检测经形态学及免疫组织化学确诊的144例石蜡包埋组织标本,包括113例B细胞淋巴瘤、24例T细胞淋巴瘤和7例淋巴结反应性增生组织.以DG75淋巴瘤细胞系DNA作为对照组.结果 引物对P1、P2、P3在B细胞淋巴瘤检出阳性率分别为71.7%(81/113),82.3%(93/113)和76.1%(86/113),三者检出率差异无统计学意义;在T细胞淋巴瘤检出率分别12.5%(3/24)、12.5%(3/24)、16.7%(4/24).将P1和P2引物组合分析,B细胞淋巴瘤阳性检出率可以达到92.3%.以上重排引物在7例反应性淋巴结中均未检出.结论 3对IgH FR3区中,新改进的P2引物在B细胞淋巴瘤中的检出率最高(82.3%).2对IgH FR3区引物联合检测可明显提高石蜡组织中B细胞淋巴瘤的检出率.     

三明地区不同年龄组女性生殖道人乳头瘤病毒不同基因类型感染状况分析

目的通过对三明地区不同年龄组女性生殖道感染人乳头瘤病毒(HPV)不同基因类型状况研究,为三明地区HPV流行病学研究提供科学依据。方法采用聚合酶多重核酸扩增(Multiplex PCR)荧光检测方法检测女性宫颈口脱落细胞进行13种高危型、5种低危型人乳头瘤病毒(HPV)检测和分型。结果在314例女性宫颈口脱落细胞标本中,HPV高危型和低危型阳性总检出率为18.78%(59/314),其中高危型检出率为12.74%(40/314);低危型检出率为6.05%(19/314)。不同年龄组:20~25岁、26~34岁、35~44岁、45~54岁、55~65岁,HPV高危型和低危型阳性总检出率分别为:40%(10/25)、13.23%(9/68)、17.18%(22/128)、22.80%(13/57)、13.88%(5/36)。其中20~25岁HPV高危型阳性检出率为20%(5/25)、HPV低危型阳性检出率为20%(5/25);26~34岁HPV高危型阳性检出率为7.35%(5/68)、HPV低危型阳性检出率为5.88%(5/68);35~44岁HPV高危型阳性检出率为11.71%(15/128)、HPV低危型阳性检出率为5.46%(7/128);45~54岁HPV高危型阳性检出率为21.05%(12/57)、HPV低危型阳性检出率为1.75%(1/57);55~65岁HPV高危型阳性检出率为8.33%(3/36)、HPV低危型阳性检出率为5.55%(2/36)。结论45~54岁组和20~25岁组的HPV高危型及20~25岁组低危型为三明地区女性生殖道人乳头瘤病毒感染的主要基因类型。     

Type-specific detection of human papillomaviruses in Kazakh esophageal squamous cell carcinoma by genotyping both E6 and L1 genes with MALDI-TOF mass spectrometry

Background: Many studies have suggested a relationship between human papillomavirus (HPV) infection and the risk of esophageal squamous cell carcinoma (ESCC). However, findings are inconclusive, potentially because of geographic heterogeneity and variations in detection methods. Objectives: We sought to further investigate the prevalence of HPV with a new detection method, the MassARRAY Sequenom technique, in esophageal squamous cell carcinomas occurring in patients belonging to Kazakh populations in Xinjiang, China. Study design: In the present study, a novel genotyping method for detecting 30 HPV genotypes, specifically by genotyping both the HPV E6 and L1 genes with multiplex PCR using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (PCR-MS) was first adopted to evaluate HPV genotypes in 89 esophageal cancer samples and 49 matched adjacent normal esophageal tissues. Results: Six HPV genotypes (HPV6, HPV16, HPV33, HPV39, HPV51, and HPV82) were present in at least 51.7% of the esophageal carcinoma tissues, which was significantly greater than 28.6% prevalence among controls (P < 0.05). HPV16 was the most common of all the genotypes investigated (HPV16 prevalence in carcinoma tissue: 49.4%; odds ratio 3.02, 95% confidence interval 1.39-6.53). HPV-positive ESCC patients were generally younger than HPV-negative patients (P = 0.04). In addition, HPV infection was more common in cases of well-differentiated and shallower invasive depth. Conclusions: Based on this new detection method, our findings reiterate the possibility that HPV infection (especially HPV16) may be involved in the etiology of esophageal carcinoma in the Kazakh populations and that HPV E6 gene positivity may be associated with prognosis of patients.     

Prevalence of cervical HPV infection in women with systemic lupus erythematosus: A systematic review and meta-analysis

Objective

The objectives of this systematic review and meta-regression were: 1) to compare the prevalence of cervical HPV infection between SLE patients and healthy controls and 2) to evaluate the relationship between cervical HPV infection and traditional and SLE-related risk factors for cervical HPV infection in these patients.

Multiplex Polymerase Chain Reaction for the Detection of High-risk-Human Papillomavirus Types in Formalin-fixed Paraffin-embedded Cervical Tissues

Detecting high-risk-human papillomavirus (HPV) types has become an integral part of the cervical cancer screening programmes. This study aimed to develop a multiplex polymerase chain reaction (PCR) for identification of HPV types 16 and 18 along with the beta globin gene in formalin-fixed and paraffin-embedded cervical biopsy specimens. A total of 59 samples from patients with cervical abnormalities were tested. HPV 16 positivity was 50% in cervical cancers and 52.9% in cervical intraepithelial neoplasia. Our multiplex PCR protocol can be used as a simple and cost-effective tool for high-risk-HPV detection in cervical cancer screening programmes.     

Comprehensive study of several general and type-specific primer pairs for detection of human papillomavirus DNA by PCR in paraffin-embedded cervical carcinomas.

We have compared the efficacies of three general primer pairs for the detection of human papillomavirus (HPV) DNA in formaldehyde-fixed paraffin-embedded carcinomas. The use of these primer pairs leads to underestimates of the HPV prevalence (GP5/6, 61.1%; CPI/IIG, 57.4%; MY09/11, 46.9%; combined, 72.8%). The efficacy of each primer pair seemed to be inversely correlated to the length of the amplimer produced. By using newly developed type-specific primer pairs (amplimer length, approximately 100 bp), an increase in HPV DNA detection (87.6%) was found.     

Prevalence of different HPV types and estimation of prognostic risk factors based on the linear array HPV genotyping test

The aim of the study was to evaluate the prevalence and risk factors of HPV in a gynecologic population attending outpatient clinics using two new molecular tests. The Amplicor HPV test and the Linear Array (LA) HPV Genotyping test were used for the detection of HPV DNA in 320 women. Multiple logistic regression was used to identify independent prognostic factors of HPV positivity. The agreement between the two methods in terms of their qualitative results was 89.3% (kappa: 0.63). Based on the LA results, the overall prevalence of HPV DNA was 49.1%, 95% confidence interval (95% CI: 43.5%, 54.7%). The prevalence of high‐risk HPV types was 30.3%. The predominant types were HPV‐6 (24.8%) and HPV‐16 (20.4%). Among women with normal cytology, the prevalence of HPV was much higher in those presenting other findings, such as inflammation, than those without other abnormal findings (49.5% vs. 31.5%). On the basis of multivariate analysis, the risk of HPV infection was higher among women with multiple sexual partners [>3 vs. 1: OR = 3.1, 95% CI: (1.5, 7.2)], Pap smear findings [low/high‐grade lesions vs. negative: OR = 2.8, 95% CI: (1.2, 6.5)], the presence of warts [yes vs. no: OR = 3.0, 95% CI: (1.5, 6.3)] and no history of child birth [no vs. yes: OR = 2.6, 95% CI: (1.0, 6.7)]. Younger age was an additional risk factor for HPV infection with carcinogenic genotypes [OR for 1 year increase = 0.93, 95% CI: (0.89, 0.98)]. J. Med. Virol. 81:2059–2065, 2009. © 2009 Wiley‐Liss, Inc.     

Human papillomavirus (HPV) DNA triage of women with atypical squamous cells of undetermined significance with Amplicor HPV and Hybrid Capture 2 assays for detection of high-grade lesions of the uterine cervix

Up to 20% of women having a cytology smear showing atypical squamous cells of undetermined significance (ASC-US) and infected with high-risk human papillomavirus (HR HPV) have high-grade cervical intraepithelial neoplasia (CIN 2/3). Results obtained with the Amplicor HPV and Hybrid Capture 2 (HC-2) assays for HR HPV DNA detection in women referred to colposcopy for an ASC-US smear were compared. Cervical samples in PreservCyt were tested for the presence of 13 HR HPV types with HC-2, with Amplicor at three cutoffs for positivity (0.2, 1.0, and 1.5 optical density units), and for 36 genotypes with the Linear Array (LA). Of 396 eligible women, 316 did not have CIN, 47 had CIN 1, 29 had CIN 2/3, and 4 had CIN of unknown grade. HR HPV was detected in 129 (32.6%) and 164 (41.4%) samples with HC-2 and Amplicor HPV (cutoff, 0.2), respectively (P = 0.01). Overall, 112 specimens were positive and 215 were negative with the HC-2 and Amplicor HPV assays (agreement of 82.6%; 95% confidence interval [CI], 78.5 to 86.0). The clinical sensitivity and specificity of Amplicor HPV at cutoffs of 0.2, 1.0 and 1.5 and of HC-2 for detection of CIN 2/3 were 89.7% (95% CI, 72.8 to 97.2) and 62.5% (95% CI, 57.5 to 52.4), 89.7% (95% CI, 72.8 to 97.2) and 64.5% (95% CI, 59.4 to 69.2), 89.7% (95% CI, 72.8 to 97.2) and 64.7% (95% CI, 59.7 to 69.5), and 93.1% (95% CI, 77.0 to 99.2) and 72.2% (95% CI, 67.4 to 76.5), respectively. Both HR HPV detection tests identified women with ASC-US who would benefit the most from colposcopy. Women with persistent HR HPV infection need further investigation despite a first normal colposcopy.     

Comparisons of HPV DNA detection by MY09/11 PCR methods

Two modifications to the original L1 consensus primer human papillomavirus (HPV) PCR method, MY09-MY011, using AmpliTaq DNA polymerase (MY-Taq), were evaluated for HPV DNA detection on clinical specimens from a cohort study of cervical cancer in Costa Rica. First, HPV DNA testing of 2978 clinical specimens by MY09-MY011 primer set, using AmpliTaq Gold DNA polymerase (MY-Gold) were compared with MY-Taq testing. There was 86.8% total agreement (kappa = 0.72, 95%CI = 0.70-75) and 69.6% agreement among positives between MY-Gold and MY-Taq. MY-Gold detected 38% more HPV infections (P < 0.0001) and 45% more cancer-associated (high-risk) HPV types (P < 0.0001) than MY-Taq, including 12 of the 13 high-risk HPV types. Analyses of discordant results using cytologic diagnoses and detection of HPV DNA by the Hybrid Capture 2 Test suggested that MY-Gold preferentially detected DNA positive specimens with lower HPV viral loads compared with MY-Taq. In a separate analysis, PGMY09-PGMY11 (PGMY-Gold), a redesigned MY09/11 primer set, was compared with MY-Gold for HPV DNA detection (n = 439). There was very good agreement between the two methods (kappa = 0.83; 95%CI = 0.77-0.88) and surprisingly no significant differences in HPV detection (P = 0.41). In conclusion, we found MY-Gold to be a more sensitive assay for the detection of HPV DNA than MY-Taq. Our data also suggest that studies reporting HPV DNA detection by PCR need to report the type of polymerase used, as well as other assay specifics, and underscore the need for worldwide standards of testing.     

Comparison of Human Papillomavirus Detections in Urine,Vulvar, and Cervical Samples from Women Attending a Colposcopy Clinic

While urine-based sampling for human papillomavirus (HPV) is being explored as a simple and noninvasive approach for cervical cancer screening, data comparing HPV genotyping in urine and those in cellular sampling of the cervix and vulva, and their correlation with rigorously confirmed cervical disease status, are sparse. We performed HPV genotyping on voided-urine and clinician-collected vulvar and cervical samples from 72 women undergoing colposcopy. Although urine-based HPV carcinogenic HPV detection was lower (58.3%) than cervical (73.6%) and vulvar (72.1%) detection (P = 0.05 and 0.07, respectively), the agreement of urine HPV with cervical and vulvar HPV was moderate (kappa = 0.55) and substantial (kappa = 0.62), respectively. Urine-based carcinogenic HPV detection had a clinical sensitivity of 80.8% (95% confidence interval [CI] = 60.7 to 93.5) and a specificity of 53.3% (95% CI = 37.9 to 68.3) for diagnosing cervical intraepithelial neoplasia grades 2/3 (CIN2/3) on histology; 90.0% of CIN3 was positive for urine HPV. The corresponding sensitivity and specificity values for vulvar sampling were 92% (95% CI = 74 to 99) and 40.5% (95% CI = 25.6 to 56.7), and those for cervical sampling were 96.2% (95% CI = 80.4 to 99.9) and 40% (95% CI = 25.7 to 55.7), respectively. HPV16 was the most common carcinogenic genotype detectable in 25% of urine, 33.8% of vulvar, and 31.9% of cervical samples overall, with prevalence increasing with cervical disease grade, regardless of the sampling method. Stronger cervical HPV PCR signal strengths were associated with increased frequency of urine HPV detection. In summary, the relatively lower detection rates but comparable clinical performance of urine-based HPV sampling underscore the need for larger studies to evaluate urine-based sampling for cervical cancer screening, epidemiologic studies, and postvaccination HPV disease surveillance.