用核酸荧光定量PCR技术检测尖锐湿疣HPV的研究

对５０例病理诊断为尖锐湿疣（ＣＡ）的标本进行了核酸荧光定量ＰＣＲ检测,检出率为９６％。结果表明：核酸荧光定量ＰＣＲ可一次性完成对尖锐湿疣ＨＰＶ感染的检测,检测覆盖面宽,漏检率低,对尖锐湿疣的诊断有较好的应用价值。     

高危型人乳头瘤病毒E7 mRNA与喉病变的关系

目的检测高危型人乳头瘤病毒(HPV)E7 mRNA表达量并分析其与喉病变的关系。方法经临床明确诊断的喉癌50例、喉癌前病变22例、声带息肉16例共计88例患者,分为喉癌组、病变组、息肉组,分别进行HPV DNA检测及HPV mRNA反转录实时荧光定量聚合酶链反应(PCR)检测。结果各组均有HPV-16及HPV-18型特异的E6、E7阳性表达。喉癌组HPV-16和HPV-18的E6及E7的相对表达量均明显高于病变组和息肉组(P<0.05);喉癌组HPV-16和HPV-18的E6及E7检出率明显高于病变组和息肉组(P<0.05);喉癌组HPV mRNA拷贝数明显高于病变组和息肉组(P<0.05),HPV mRNA的表达水平随喉病变的发展而提高。结论 HPV mRNA的表达率随喉病变的发展而增加,其表达量也随之增强,其表达水平与喉病变发展具有一定相关性。检测HPV E7 mRNA可以为喉癌的预防、发展监测提供重要的临床指导。     

人乳头瘤病毒快速检测分型方法的临床应用

目的探讨人乳头瘤病毒(HPV)免疫层析技术快速检测分型方法的临床应用价值。方法采用免疫层析技术快速检测法对人乳头瘤病毒进行检测分型,采用基因检测技术PCR检测法作对照。结果对386例标本进行HPV常见亚型6、11型和16、18型检测,快速检测法与对照PCR检测法均105例阳性,阳性率均为27.20%,两种方法的阳性率一致(P>0.05)。对HPV亚型总的分型检测,快速检测法检出阳性105例,阳性率为27.20%;对照PCR检测法检出HPV阳性142例,阳性率为36.78%,两种方法的阳性率差异有统计学意义(χ2=8.15,P<0.05)。结论免疫层析快速检测分型方法用于HPV常见亚型6、11型(低危型)和16、18型(高危型)检测,具有操作简便,检测速度快,准确率高,医疗成本较低等特点,值得基层医院推广应用。     

110例尖锐湿疣患者FQ-PCR检测结果分析

目的 探讨尖锐湿疣患者人乳头瘤病毒(HPV)感染分型及其意义. 方法 采用荧光定量PCR技术(FQ-PCR)检测男女尖锐湿疣患者HPV感染分型. 结果 男性患者44例,检出阳性37例(84.09%),分型结果以HPV6/11型为主(91.89%);女性患者外阴标本33例,检出阳性26例(78.78%),分型结果以HPV6/11型为主(53.84%);宫颈标本33例,检出阳性19例(57.57%),分型结果以HPV6/11型为主(57.89%). 结论 HPV6/11型是尖锐湿疣发病的主要型别,荧光定量聚合酶链反应(FQ-PCR)可作为尖锐湿疣诊断的指标.     

尖锐湿疣患者荧光定量PCR检测结果分析

目的探讨尖锐湿疣患者疣体中人乳头瘤病毒(HPV)分型、基因含量及其意义。方法采用荧光定量PCR技术(FQ-PCR)检测尖锐湿疣患者疣体中HPV-DNA含量及其型别。结果169例尖锐湿疣患者经FQ-PCR检测后168例(99.41%)阳性,其中HPV6/11型阳性150例(88.76%),平均1.2×107拷贝/ml;HPV16/18型阳性13例(7.69%),平均2.3×106拷贝/ml;HPV6/11和HPV16/18型同时阳性5例(2.96%),平均1.0×107拷贝/ml;4型均阴性1例(0.59%),定量为0×10拷贝/ml。结论HPV6/11型是尖锐湿疣发病的主要型别,尖锐湿疣患者疣体中HPV-DNA含量与其是否复发无明显关系,FQ-PCR可作为尖锐湿疣早期诊断的指标。     

老年喉癌组织中人乳头状瘤病毒E7及肿瘤增殖、迁移相关基因mRNA表达量的检测及临床意义

目的检测老年喉癌组织中增殖、迁移相关基因及人乳头状瘤病毒(HPV)E7 mRNA表达量并分析其临床意义。方法经临床明确诊断的喉癌、喉癌前病变、声带息肉患者共计88例,分为喉癌组、病变组、息肉组3组,分别扩增目的基因丝氨酸丰富剪接因子(SRSF)1、重组人微管解聚蛋白(STMN)1、结肠癌转移相关基因(MACC)1、生存素(Survivin)、β-链蛋白(catenin)、E-钙黏蛋白(cadherin)、N-cadherin、红细胞波形蛋白纤维(Vimentin)、CD44v6、基质金属蛋白酶(MMP)-9、含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-1、Beclin1及内参基因GAPDH,计算其mRNA含量。结果喉癌组HPV-16及HPV-18的E7 mRNA含量均显著高于病变组和息肉组(P<0.05);喉癌组织中增殖基因SRSF1、STMN1、MACC1、Survivin的mRNA含量显著高于病变及息肉组(P<0.05);喉癌组织中E-cadherin、β-catenin的mRNA含量显著低于病变组和息肉组(P<0.05);喉癌组织中Ncadherin、Vimentin、CD44v6、MMP-9的mRNA含量显著高于病变组和息肉组(P<0.05);喉癌组织中Caspase-1、Beclin1的mRNA含量均显著低于病变组和息肉组(P<0.05)。结论喉癌组织中HPV mRNA表达量异常升高,并与喉癌细胞增殖、迁移相关基因表达量相关。检测HPV E7 mRNA可以为喉癌的预防、发展监测提供重要的临床指导。     

宫颈癌盆腔淋巴结HPV DNA检测的临床意义

以宫颈癌盆腔淋巴结中高危型人乳头状瘤病毒16/18(HPV16/18)特异性引物通过PCR方法,检测27例宫颈癌477枚盆腔淋巴结中HPV16/18 DNA.27例宫颈癌原发灶HPV16/18 DNA检出21例,21例HPV16/18阳性原发灶的369枚淋巴结阳性率为13.82%.淋巴结HPV DNA阳性率在不同年龄、肿瘤直径、细胞分化程度、病理类型、肌层浸润深度、有无淋巴结转移和不同淋巴结清扫数的宫颈癌差异有显著性意义.认为在无转移淋巴结检测到HPV基因,对宫颈癌微小转移具有提示意义,有助于发现普通病理学难以检测到的早期隐匿性转移.     

应用荧光定量PCR技术检测HBV DNA低拷贝样品及临床意义分析

目的：观察荧光定量PCR技术对HBVDNA低拷贝样品检测的情况并探讨其临床意义。方法：采用荧光定量PCR和改良PCR法对一系列不同拷贝数的临床标本平行进行检测。结果：在电泳分析时PCR产物的亮度与样品的拷贝数呈正相关，〉10^3copies／ml的样品能经PCR扩增得到明亮的特异产物条带，10^3copies／ml的样品也可得到特异的弱条带，但10^2copies／ml的样品无法检测到特异产物，并有明亮的引物聚合体产生。结论：荧光定量PCR技术的检测准确下限为10^4copies／ml，10^3copies／ml的样品检测时存在很大的随机性，这种条件下的样品并不能用荧光定量PCR的方法进行准确检测，应该采用其他更灵敏的方法。     

尖锐湿疣患者皮损中人乳头瘤病毒检测与临床复发关系的研究

目的探讨杭州地区尖锐湿疣患者中人乳头瘤病毒(HPV)感染的型别,以及不同病毒型别和病毒载量与复发的关系。方法采用实时荧光定量聚合酶链反应(FQ-PCR)检测了289例患者皮损中的HPV,并作临床跟踪观察3个月,记录复发次数。结果289例患者中仅HPV6/11型感染214例,占74.04%;37例为HPV6/11和HPV 16/18混合感染,占12.80%:仅HPV16/18阳性者16例,占5.54%;HPV6/11和16/18均阴性者22例,占7.61%。53例HPV16/18阳性感染者中,男18例,阳性率为13.14%;女35例,阳性率为23.03%,差异有统计学意义(χ2=4.6979,P<0.05)。HPV16/18型感染组与HPV6/11型感染组的3个月复发率差异无统计学意义(P>0.05),但复发次数高于HPV6/11感染组(P<0.05)。不同HPV6/11型病毒载量组之间的3个月复发率和复发次数差异均无统计学意义(P>0.05)。结论尖锐湿疣患者的HPV感染型别以6/11型为主,女性患者中高危型HPV16/18型的感染率明显高于男性。不同HPV型别与复发率无关,但高危型HPV感染者复发次数增加。不同HPV6/11型病毒载量与尖锐湿疣的临床复发无关。     

人类乳头瘤病毒HPV16,18型感染与胃癌发生的关系

目的探讨人类乳头瘤病毒ＨＰＶ１６,１８型感染与胃癌发生的关系．方法采用ＨＰＶ１６Ｅ６和ＨＰＶ１８Ｅ６／Ｅ７特异性引物,通过多聚酶链反应（ＰＣＲ）,检测３７例新鲜胃癌组织标本及２０例新鲜癌旁正常胃粘膜标本中的ＨＰＶＤＮＡ．结果新鲜胃癌组织３７例中ＨＰＶ１６,１８型ＤＮＡ检出率分别为２１６％和５４％．新鲜癌旁正常胃粘膜标本２０例中检出率为０．ＨＰＶ１６型ＤＮＡ在胃癌和癌旁正常胃粘膜中检出率存在显著性差异（Ｐ＜００５）,而ＨＰＶ１８则没有显著性差异（Ｐ＞００５）．新鲜胃癌组织３７例中ＨＰＶ１６,１８型ＤＮＡ总检出率为２７０％,２０例新鲜癌旁正常胃粘膜标本中总检出率为０,两者有显著性差异（Ｐ＜００５）．新鲜胃癌组织３７例中ＨＰＶ１６,１８型双重感染检出率为０．ＨＰＶ１６和（或）ＨＰＶ１８型ＤＮＡ的检出率在不同分化程度的胃癌中无显著性差异,但ＨＰＶ１６型ＤＮＡ在不同部位的胃癌中检出率却有显著性差异,以贲门部胃癌检出率最高．结论高危类ＨＰＶ１６型感染可能与胃癌发生有关,ＨＰＶ１６型病毒癌基因可能在胃癌形成过程中起一定作用     

新疆哈萨克族食管癌中HPV感染状况分析

目的:探讨人乳头状病毒(HPV)18, 31, 45E7 3个高危基因亚型在新疆哈萨克族(哈族)食管癌中的感染率及其相关性. 方法:采用聚合酶链反应(PCR)技术检测HPV-18, 31, 45E7基因在316例哈族食管癌中的感染情况. 并随机抽取PCR产物进行测序来验证PCR结果.结果:HPV18, 31和45E7基因亚型在食管癌中阳性率分别为25.3%(80/316), 14.2%(45/316)和7.3%(23/316), 其基因型分布与年龄、性别均无关. HPV31, 45E7基因阳性率在组织病理学分级上无统计学意义. HPV18基因型分布在中低分化食管鳞状细胞癌中的检出率明显高于高分化食管鳞癌(29.6% vs 17.7%, χ2 = 5.398, P<0.05).结论:HPV18型是新疆哈族食管癌的主要高危亚型之一, 其基因型分布与食管癌分化程度有关;HPV31, 45亚型可能不是新疆哈族食管癌最主要的高危亚型.     

人乳头瘤病毒16与胃癌相关性的研究

目的探讨人乳头瘤病毒１６（ＨＰＶ１６）与胃癌发生的关系。方法用聚合酶链式反应（ＰＣＲ）扩增出ＨＰＶ１６早期区Ｅ６的１２０ｂｐＤＮＡ片段，扩增产物与５＇末端３２Ｐ标记的特异性寡核苷酸探针进行斑点杂交及放射性自显影，用此方法对３４６份新鲜的及福尔马林固定石蜡包埋的胃组织标本中ＨＰＶ１６ＤＮＡ进行检测。结果胃腺癌、癌旁粘膜、胃局部淋巴结及正常胃粘膜组织中ＨＰＶ１６的检出率分别为：２２．７２％（３０／１３２）、５．８８％（４／６８）、０％（０／４２）、２．８８％（３／１０４），胃癌组ＨＰＶ１６的检出率高于其他各组，统计学上差异均有显著性（Ｐ＜０．０１）。结论ＨＰＶ１６可感染胃粘膜上皮细胞，可能是胃癌的致癌因素之一。     

Anal cancer in Chinese: human papillomavirus infection and altered expression of p53

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Anal cancer in Chinese: human papillomavirus infection and altered expression of p53

AIM To detect the presence of HPV DNA and study the alteration of p53 expression in anal cancers in Chinese.METHODS HPV DNA was amplified by PCR. The amplified HPV DNA was classified by DBH. HPV antigen and p53 expression were respectively detected by immunohistochemistry.RESULTS HPV DNA was amplified only in one case of squamous cell carcinoma of the 72 Chinese anal cancers and further classified as HPV type 16. Others were all HPV negative. HPV antigen and p53 expression were also detected in this case. Positive stainings with anti-p53 antibody were seen in 61.2% anal cancers. There were no statistically significant differences between anal squamous cell carcinomas and adenocarcinomas and between anal adenocarcinomas and rectal adenocarcinomas. p53 protein expression was observed in the basal cells of squamous epithelium of condyloma acuminatum and morphologically normal squamous epithelium in 2 cases invaded by anal adenocarcinoma.CONCLUSION HPV infection was not associated with these cases of anal cancer. p53 alteration was a common event. Positive p53 immunostaining can not be regarded as a marker for differentiating benign from malignant lesions.     

Identification of human papillomavirus as a preventive strategy for cervical cancer in asymptomatic women in the Peruvian Andes

ObjectiveTo detect the most prevalent human papillomavirus (HPV) genotypes in cervical smear samples of asymptomatic Peruvian women by analyzing the correlation between Papanicolaou (PAP)-stained cervical tests and PCR-sequencing.MethodsA total of 254 women attending routine gynecological examinations were included in this study. The samples were analyzed by PAP technique and examined under a microscope by a pathologist and classified by the Bethesda system. HPV amplification was done using the primers specific for E1 region and positive specimens were confirmed by direct sequencing.ResultsThe prevalence of HPV was investigated in 254 cervical scrape samples by PCR. PAP smear showed that 94.9% cases had normal morphology and 5.1% had an inflammatory pattern; 20.5% were found to be infected with HPV, comprising 20 different genotypes. HPV16 was the most prevalent genotype in correlation with changes in cervical cytology.ConclusionsOur results suggest the HPV is very frequent even in women with negative PAP, and PCR seems to be the best option to determine the causative agent of HPV infection in endocervical samples. Identification of the HPV genotype in asymptomatic women may allow the implementation of appropriate prophylactic measures which may have a direct impact on the natural history of the disease and the subsequent development of cervical malignancy.     

Detection of human papillomavirus DNA in esophageal carcinoma in Greece

AIM:To detect human papillomavirus(HPV)in theesophageal mucosa and the possible relationship with esophageal cancer in Greece.METHODS:Forty-nine patients underwent esophagogastroduodenoscopy(EGD)and esophageal biopsy at a university hospital that acts as a referral center for Northern Greece.Nineteen of these patients(14 male and 5 female)had esophageal squamous cell carcinoma(ESCC)and 30(15 male and 15 female)did not have any reported esophageal malignancy.Histopathological assessment was followed by polymerase chain reaction analysis of all the samples.Patient demographic data(age,sex,and place of birth)and information regarding smoking habits,alcohol consumption or sexual habits were collected.A method of statistical interference,verification of hypotheses based on homogeneity and independentχ2 test,was used.RESULTS:From the 49 patients that underwent EGD and biopsy,19 had ESCC and 30 had normal esophageal mucosa,with a mean age of 65.2 years.Regarding the prevalence of oncogenic risk factors for esophageal carcinoma,an interesting conclusion was that 78%of the patients used tobacco and almost one-third had multiple sexual partners,whereas only 20%of the patients consumed alcohol,which was not statistically significant,when compared to the control group.In the ESCC group,the only two positive samples were among the male patients(2/14 male patients with ESCC,14.5%).No HPV was identified in the control group.The predominant HPV types identified were 11 and 31,which have a low malignancy potential.The presence of HPV DNA in the ESCC group was not statistically significant,95%confidence interval(χ2=3.292,P=0.07).CONCLUSION:This is the first relevant study in Greece,and despite the lack of statistical significance,the issue of HPV infection and ESCC does merit further investigation.     

Coinfection of Chlamydia trachomatis, Ureaplasma urealyticum and human papillomavirus among patients attending STD clinics in Estonia

Women visiting Estonian STD clinics were subjected to PCR assay for human papillomavirus (HPV), Chlamydia trachomatis and Ureaplasma urealyticum biovar 2. The overall prevalence of coinfection was 8%. The chlamydial infection was found to be associated with HPV, especially with high-risk HPV (OR=2.5, p<0.005) and most significantly in women over 41 y of age. C. trachomatis infection also occurred more frequently in U. urealyticum-infected than in U. urealyticum-free patients (OR=2.6, p=0.02). U. urealyticum infection did not associate with HPV status. The clinical significance of the association between C. trachomatis and U. urelyticum infection remains to be elucidated.     

Prevalence of human papillomavirus in esophageal carcinoma in Tangshan,China

AIM:to study the prevalence of human papillomavirus(HPV) in esophageal carcinoma in tangshan,China,a high-incidence area.METHODS:Formalin-fixed,paraffin-embedded tissue specimens from 198 patients who were pathologically diagnosed with esophageal squamous cell carcinoma from 2011 to 2013 were obtained from a pathology department in Tangshan.DNA was extracted from all198 specimens to detect HPV by polymerase chain reaction(PCR).β-globin PCR was performed to check the quality of the DNA extraction procedure.PCR was performed to detect a wide range of HPV types,and type-specific PCR was performed to detect HPV types16 and 18.Negative and positive controls were used for HPV 16 and 18 detection.RESULTS:the DNA extraction method in this study appeared to be more effective than other previously reported methods.After DNA extraction,more than98%of the tissue specimens had an acceptable result in the DNA qualification test(β-globin PCR).the overall prevalence of HPV in tumor tissues by GP6+/GP5+PCR was 79.79%,and the prevalence of HPV types16 and 18 was 40.40%and 47.47%,respectively.PCR demonstrated the presence of HPV,and direct sequencing confirmed the HPV genotypes.All HPVpositive PCR products were checked by DNA sequence analysis using DNAman and compared with the known HPV sequences listed in the basic Local Alignment Search tool database to evaluate the HPV types.this analysis confirmed the presence of HPV types 16 and18.CONCLUSION:DNA of high-risk HPV types 16 and 18is present in esophageal tumors,implicating HPV as a possible etiologic factor for esophageal squamous cell carcinoma.