用聚合酶链反应检测食管癌组织中人乳头瘤病毒DNA

应用聚合酶链反应（ＰＣＲ）技术对汕头市区６８例食管癌的石蜡包埋标本进行人乳头瘤病毒（ＨＰＶ）ＤＮＡ序列检测，结果显示，ＨＰＶＤＮＡ总阳性率为６６．１８％（４５／６８），检出型别主要为ＨＰＶ６、１１、１６，检出率分别为２７．９４％、３６．７６％和２７．９４％，经统计学处理三型间无显著性差异；ＨＰＶ－１８及未定型别各占８．８２％。值得注意的是ＨＰＶ感染中多重感染占阳性病例的５３．３３％（２４／４５）。初步结果表明，汕头市食管癌高发区有较高的ＨＰＶ感染率，此与食管癌的发生，可能有密切关系。     

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.     

应用通用引物聚合酶链反应技术检测结肠癌组织中人乳头瘤病毒DNA

应用人乳头瘤病毒（ＨＰＶ）通用引物介导的聚合酶链反应（ＰＣＲ）技术检测了１５例结肠癌石蜡包埋病理组织切片中ＨＰＶＤＮＡ，其中１０例呈阳性扩增（阳性率为６６．７％）。１２例正常结肠组织经上述ＰＣＲ检测均呈阴性反应。阳性扩增产物经核酸斑点杂交进行ＨＰＶ型别分析，ＨＰＶ１６型占４例（４０．０％），１８型１例（１０．０％），１６／１８型５例（５０．０％），未检出其他ＨＰＶ型别。表明ＨＰＶ可能对结肠癌的发生具有病原相关性。     

用PCR检测肺癌组织中人乳头瘤病毒16、18型DNA相关序列

用ＰＣＲ检测肺癌组织中人乳头瘤病毒１６、１８型ＤＮＡ相关序列胡克，李清泉，潘显光，曹作炎，杨炯人乳头瘤病毒（ＨＰＶ）与人类某些恶性肿瘤的发生有密切关系，ＨＰＶ可能在病因学方面参与机体多部位肿瘤的形成。作者应用ＰＣＲ技术检测肺癌组织中ＨＰＶ１６、１８型...     

尖锐湿疣组织中人乳头状瘤病毒的检测

用免疫组织化学、ＤＮＡ原位杂交和聚合酶链反应技术，检测人生殖器尖锐湿疣和女阴假性湿疣组织中人乳头状瘤病毒衣壳抗原（ＨＰＶ－Ａｇ）和病毒核酸序列（ＨＰＶ－ＤＮＡ），并观察了ＨＰＶ在尖锐湿疣组织中的分布特点与病变组织学改变的关系。结果显示尖锐湿疣中ＨＰＶ－Ａｇ阳性率为７１．４％（３５／４９）；原位杂交ＨＰＶ６／１１ＤＮＡ阳性率为９６．５％（２８／２９）；ＰＣＲ扩增后尖锐湿疣ＨＰＶ６／１１／１６／１８ＤＮＡ阳性率为１００％（５３／５３）；假性湿疣ＨＰＶ６／１１／１６／１８ＤＮＡ阳性率为２１．４％（３／１４）。观察ＨＰＶ－Ａｇ和ＨＰＶ－ＤＮＡ分布，表明ＨＰＶ增殖性感染和尖锐湿疣特异的病理改变密切相关。     

子宫颈组织中人乳头瘤病毒16型E6基因片段的定量PCR检测

目的探讨子宫颈组织中人乳头瘤病毒(HPV)16型E6基因的含量与疾病严重程度的关系.方法用定量聚合酶链反应(PCR)检测20例慢性宫颈炎,6例宫颈非典型增生,18例宫颈癌组织中HPV16E6基因的拷贝数.结果HPV16E6基因在慢性宫颈炎,宫颈非典型增生及宫颈癌组织中的平均拷贝数(拷贝/μgDNA)分别为6.16×104,5.33×106和6.45×106.统计学处理表明,宫颈非典型增生及宫颈癌组织中E6基因的拷贝数显著高于慢性宫颈炎(P<0.01),宫颈癌组织中E6基因的拷贝数高于宫颈非典型增生组织,但差异无显著性(P>0.05).结论HPV16E6基因的拷贝数与宫颈疾病程度呈正相关,定量检测HPV16E6基因可作为监测宫颈癌高危人群的一种方法.     

人喉癌组织中人乳头瘤病毒DNA的检测

目的为探讨喉癌与人乳头瘤病毒（ＨＰＶ）感染的关系和ＨＰＶ在喉癌中基因组型的分布与表达。方法应用聚合酶链反应技术（ＰＣＲ）制备非放射性探针标记物－地高辛标记ＨＰＶ共有引物探针,对１４６例喉不同病变的新鲜组织标本（喉癌６８例,喉其它病变４８例,正常喉组织３０例）,进行ＨＰＶ６,１１,１６,１８,３１,３３,３５,４２,５８共９型ＨＰＶＤＮＡ感染的检测;阳性者用多重引物ＰＣＲ方法分型。结果喉癌ＨＰＶ感染阳性率４５．６％（３１／６８）,喉癌颈转移淋巴结组织阳性率２０．０％（３／１５）,喉癌前病变阳性率１１．８％（２／１７）,声带息肉阳性率６．３％（１／１６）,１５例癌旁及１５例癌周正常喉组织均为ＨＰＶＤＮＡ阴性。ＨＰＶＤＮＡ型别分布在喉癌中以ＨＰＶ１６、１８型为主,喉良性病变中以ＨＰＶ６、１１型为主。结论喉癌发生与ＨＰＶ感染有关。     

宫颈透明细胞癌中人乳头瘤病毒的检测

目的 探讨宫颈透明细胞癌中高危型HPV感染情况.方法 提取1例37岁宫颈透明细胞癌患者手术切除组织蜡块中的DNA,通过巢式PCR方法检测其中HPV感染情况.结果 该患者肿瘤切除组织高危型HPV18型阳性.结论 利用巢式PCR方法分型检测宫颈透明细胞癌中的高危型HPV型别,其准确性及敏感性均较高.     

应用原位PCR技术检测阴茎癌组织中人乳头瘤病毒DNA

目的：研究人乳头瘤病毒（ＨＰＶ）与阴茎癌的关系。方法：应用原位ＰＣＲ技术对４６例阴茎癌组织中的ＨＰＶ１６和ＨＰＶ１８ＤＮＡ进行检测。结果：３３例阴茎癌中检测到了ＨＰＶＤＮＡ（７１．７％），其中２９例ＨＰＶ１６ＤＮＡ阳性（６３．０％），６例ＨＰＶ１８ＤＮＡ阳性（１３．０％），２例ＨＰＶ１６和ＨＰＶ１８ＤＮＡ均阳性，４例ＨＰＶ１８ＤＮＡ阳性而ＨＰＶ１６阴性；２例淋巴结转移癌，３例癌旁不典型增生组织和１例癌旁增生组织ＨＰＶ１６ＤＮＡ阳性。结论：阴茎癌与ＨＰＶ１６、ＨＰＶ１８感染有密切关系，原位ＰＣＲ技术是一项敏感性高、特异性强的技术。     

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。     

Detection of human papillomavirus DNA in fine-needle aspirations of metastatic squamous-cell carcinoma of the uterine cervix using the polymerase chain reaction

Routinely processed fine-needle aspirations of metastatic squamous-cell carcinoma (SCC) were analyzed for human papillomavirus type 16 (HPV-16) using the polymerase chain reaction (PCR), an in vitro DNA amplification method. HPV-16 DNA was detected in five of seven (71%) metastases from SCC of the uterine cervix. In two cases in which the primary tumor was available for comparison, the HPV-16 DNA content of the primary tumor and of the metastasis was identical. HPV-16 was not found in a metastatic SCC from the lung or in a metastatic nasopharyngeal SCC. These findings demonstrate that HPV-16 DNA sequences can be readily detected in routinely processed fine-needle aspirations using the polymerase chain reaction. The finding of HPV-16 DNA in a metastasis may serve to direct a search for a primary site of origin.     

妊娠期人乳头瘤病毒的感染状况及母婴传播的研究

应用聚合酶链反应检测孕妇尖锐湿疣（ＣＡ）组织、无ＣＡ孕妇宫颈分泌物、外周血及羊水、新生儿咽分泌物、脐血中人乳头瘤病毒（ＨＰＶ）感染及母婴传播。结果表明：ＣＡ组织中ＨＰＶＤＮＡ阳性率为９０．３２％，以ＨＰＶ６／１１型为主；无ＣＡ孕妇宫颈分泌物中ＨＰＶＤＮＡ阳性率为３５．７１％，以ＨＰＶ１６／１８型为主；孕妇血中ＨＰＶＤＮＡ阳性率为５７．６９％；母婴间经产道垂直传播率为４４．４４％，血性经胎盘传播传播率为６０％。说明ＨＰＶ不仅存在于ＣＡ组织中，还存在于无ＣＡ孕妇生殖道及外周血中，母婴间传播途径除产道外，还有胎盘传播     

宫颈癌组织中人乳头瘤病毒16型E7蛋白致癌机理初探

目的 研究宫颈癌组织中人乳头瘤病毒（ＨＰＶ）１６－Ｅ７蛋白对视网膜母细胞瘤基因（Ｒｅｔｉｎｏｂｌａｓｔｏｍａ）Ｒｂ蛋白及Ｅ２Ｆ－１的作用的机制,探讨ＨＰＶ１６－Ｅ７蛋白与宫颈癌发生的关系。方法 采用聚合酶链反应检测宫颈癌及正常宫颈组织中ＨＰＶ１６感染等,用蛋白印迹技术对ＨＰＶ１６ ＤＮＡ阳性的宫颈癌组织中是否存在ＨＰＶ１６－Ｅ７蛋白和Ｒ６蛋白－Ｅ２Ｆ－１形成的复合物进行检测。正常宫颈组织作为对照,     

子宫颈刮片中人乳头瘤病毒的基因分型

目的 确定不同型人乳头瘤病毒（ＨＰＶ）感染的自然历程以及其持续感染在子宫颈癌发展过程中的作用。方法 应用聚合酶链反应检测荷兰１５５名妇女子宫刮颈片中的ＨＰＶ ＤＮＡ，应用线样探针分析法（ＬｉＰＡ）进行，包括ＨＰＶ６，１１，１６，１８，３１，３３，３５，４０，４２，４３，４４，４５，５１，５２，５６和５８的基因分型。结果 １５５例妇女子宫颈片中ＨＰＶ ＤＮＡ检出率为６０％，其中在宫颈细胞学检查正常或     

Expression of p53 protein in laryngeal squamous cell carcinoma and dysplasia: possible correlation with human papillomavirus infection and clinicopathological findings

In order to evaluate the expression of p53 protein in 28 premalignant and 40 malignant squamous cell proliferations of the larynx and its relationship to tobacco consumption, human papillomavirus infection and differentiation grade of the lesions, p53 expression was examined by means of a microwave post-fixation immunohistochemical method using the PAb 240 and PAb 1801 monoclonal antibodies. HPV infection was assessed by non-isotopic in situ hybridization (NISH) and polymerase chain reaction (PCR). A large proportion of carcinomas (77.5%) and dysplasias (61%) expressed p53. No difference was found between differentiation grades of the lesions regarding p53 detection (P>0.1), but moderate or intense p53 expression was more frequent in the carcinomas (P<0.05). A statistical correlation was found between cigarette consumption and both p53 detection and p53 staining intensity (P<0.05 in each case). HPV study revealed HPV 16 and 18 infection only in carcinomas. The frequency was 28% and the physical state of the virus as demonstrated by NISH was integration into the genome. We observed an inverse relationship between HPV infection and p53 expression (P=0.006). Our findings suggest that p53 overexpression is a common and early event which increases in frequency with progression of laryngeal squamous cell carcinoma. The expression of p53 is influenced by tobacco and high-risk types of HPV.     

慢性宫颈炎患者宫颈中人乳头瘤病毒基因的检测

应用聚合酶链反应技术检测正常妇女与慢性宫颈炎患者宫颈人乳头瘤病毒感染情况的结果显示，南京市慢性宫颈炎患者ＨＰＶ－ＤＮＡ总阳性率达５８．６％；ＨＰＶ６，１１，１６，１８型阳性率分别为２９．３％，３０．７％，２８．６％和３４．３％，而正常妇女中ＨＰＶ总阳性率为１９．４％，４个型别ＨＰＶ阳性率为９．７％，１６．１％，３．２％和３．２％，明显低于宫颈炎患者。如皋市慢性宫颈炎患者ＨＰＶ－ＤＮＡ总阳性率高达８     

Rapid real time PCR to distinguish between high risk human papillomavirus types 16 and 18

Aims—To assess the validity and practicality of real time polymerase chain reaction (PCR) for human papillomavirus (HPV) testing in combination with liquid based cytology samples for cervical screening.Methods—Real time PCR using consensus (GP5+/6+) and type specific primers was developed to detect genital HPV types. This provides rapid, efficient amplification followed by denaturation of the product and computer analysis of the kinetics data that are generated. Liquid based cytology samples were obtained from patients attending routine cervical screening clinics. DNA was extracted from the residual cellular suspension after cytology using spin columns.Results—Real time PCR successfully distinguished between HPV-16 and HPV-18 on the basis of amplification with consensus primers followed by DNA melting temperature (T_m) analysis. Sensitivities of one to 10 copies of HPV-16 (mean T_m = 79.4°C; 2 SD, 0.8) and four to 40 copies of HPV-18 (mean T_m = 80.4°C; 2 SD, 0.4) were obtained. In a mixed population of SiHa and HeLa cells containing known copy numbers of HPV-16 and HPV-18 genomes, HPV-16 and HPV-18 products were clearly separated by T_m analysis in mixtures varying from equivalence to 1/1000. Together with detailed melt analysis, type specific primers from the same region of the L1 gene confirmed the differential ability of this system. The method was applied to 100 liquid based cytology samples where HPV status using conventional GP5+/6+ PCR was already known. There was 95% agreement between the methods, with 55 positives detected by conventional PCR and 59 with real time PCR. The method was then tested on 200 routine liquid based cytology samples. Approximately 10% were positive by real time PCR, most of which were classified as HPV-16 by detailed melt analysis. Thirteen (6.8%) HPV positives were identified in 189 samples showing no evidence of cervical cytological abnormality.Conclusions—Real time PCR is a rapid, efficient method for the detection of HPV with the separation of HPV-16 and HPV-18 on the basis of differential T_m. Preliminary results suggest it could prove useful if HPV testing is added to cervical screening programmes.     

Prognostic value of human papillomavirus types 16 and 18 DNA physical status in cervical intraepithelial neoplasia

The aim of this work was to assess the value of the physical status of human papillomavirus (HPV) DNA as a disease marker for cervical cancer development in a set of 248 DNA samples previously genotyped as HPV 16 or 18, by calculating the E2/E6 ratio through real-time PCR. There was a significant difference in integration status according to disease grade for both genotypes (p <0.001). Furthermore, especially for HPV 18, determining the DNA physical status could be a useful biomarker in predicting cervical cancer risk development, with a lower E2/E6 ratio clinically associated with the development of a precancerous lesion.