DIG—DNA探针检测宫颈癌组织中人乳头瘤病毒6,11,16,18型DNA

用地高辛配基分别标记，制备ＨＰＶ－６、ＨＰＶ－１１、ＨＰＶ－１６及ＨＰＶ－１８的ＤＮＡ探针，应用核酸杂交技术，对３７例宫颈癌活检组织ＤＮＡ作了检测，各型ＨＰＶＤＮＡ的检出率为：ＨＰＶ－６０％、ＨＰＶ－１１２．７、ＨＰＶ－１６６７．５％、ＨＰＶ－１８１３．５％，联合应用ＨＰＶ－１６和ＨＰＶ－１８探针共同检测结果为８１％。结果显示：我国宫颈癌的发生与ＨＰＶ感染密切相关，联合应用高危型ＨＰＶ探针可提高检     

子宫颈组织中人乳头状瘤病毒的PCR检测

①目的探讨人乳头状瘤病毒１６，１８型（ＨＰＶ－１６，ＨＰＶ－１８）感染与慢性宫颈炎、宫颈癌的关系。②方法聚合酶链反应（ＰＣＲ）技术检测２２例正常宫颈、５５例慢性宫颈炎、３０例宫颈癌的子宫颈组织。③结果正常宫颈组织中ＨＰＶ－１６，１８的检出率均为０．００％，慢性宫颈炎分别为１８．１８％和７．２７％，宫颈癌分别为５６．６７％和３．３３％．统计学处理表明，宫颈癌组织中ＨＰＶ－１６的检出率明显高于宫颈炎组织（χ２＝１３．６２４，Ｐ＜０．０１），而且Ⅲ度宫颈糜烂组织中ＨＰＶ－１６的检出率也较Ⅰ，Ⅱ度的检出率高，两者比较差异有显著意义（χ２＝１０．５５０，Ｐ＜０．０１），但不同宫颈疾患组织中ＨＰＶ－１８的检出率无统计学意义。④结论子宫颈癌的发生与ＨＰＶ感染密切相关，以ＨＰＶ－１６尤为明显，ＰＣＲ技术是检测组织中ＨＰＶ－ＤＮＡ的敏感、特异的方法     

双温聚合酶链反应检测喉乳头瘤中HPV－DNA

应用双温聚合酶链反应（ＰＣＲ）检测喉乳头瘤石蜡包埋组织中的人乳头瘤病毒（ＨＰＶ）ＤＮＡ。结果与常规ＰＣＲ（三温循环）相同：３０例喉乳头瘤中ＨＰＶ总检出率为４６．７％（１４／３０），ＨＰＶ－６／１１／１６的检出率分别为４０％（１２／３０）、１６．７％（５／３０）和６．７％（２／３０），ＨＰＶ－６的检出率极显著高于ＨＰＶ－１１／６（ｘ２＝１０．６，Ｐ＜０．０１）。初步结果表明，本地区喉乳头瘤主要与ＨＰＶ－６的感染密切相关。该法具有缩短检测时间、节省一个恒温水浴箱等优点。     

广谱人乳头瘤病毒DNA探针对尖锐湿疣和假性湿疣的原位杂交检测

对用肉眼形态、组织学特征和人乳头瘤病毒核壳抗原（ＨＰＶ－Ａｇ）来检测诊断为尖锐湿疣和假性湿疣的两组病例，用广谱人乳头瘤病毒ＤＮＡ探针，作核酸原位杂交检测，结果：尖锐湿疣组ＨＰＶ－ＤＮＡ检出率为３０／３３（９０．９％），而假性湿疣组检出率仅为１／３８（２．６％）（Ｐ＜０．０００１），提示假性湿疣并非为ＨＰＶ感染引起，或ＨＰＶ感染的可能性很小。作者还对应用广谱人乳头瘤病毒ＤＮＡ原位杂交作为对疑为ＨＰＶ感染病变作病理学初筛检测的意义作了讨论。     

食管癌组织中人乳头瘤病毒的检测

用人乳头瘤病毒（ＨＰＶ）保守基因的通用引物和１６、１８型特异性引物，通过聚合酶链反应（ＰＣＲ），检测２０例正常新鲜活检食管粘膜，４０例新鲜食管癌组织和５１例石蜡包埋的食管癌组织标本的ＨＰＶＤＮＡ，在２０例正常人食管粘膜组织中ＨＰＶＤＮＡ检出率９．１％，癌组织中为３６．２％，鳞癌和腺癌分别为３４．７％、４７．１％，鳞癌以１６型感染为主，腺癌以１８型为主。但肿瘤分化程度与ＨＰＶ感染无关。结果提示：ＨＰＶ感染与食管癌的发生有一定关系，不同型别的ＨＰＶ与不同组织的亲和性有差别。     

乳头瘤病毒及巨细胞病毒在宫颈癌组织中的协同作用

应用ＤＮＡ－ＤＮＡ斑点杂交技术，对６４例宫颈标本（包括正常宫颈２６例，重度宫颈糜烂１７例，宫颈癌２１例）分别检测ＨＰＶ－１６ＤＮＡ及ＨＣＭＶＤＮＡ。结果发现正常宫颈、宫颈糜烂及宫颈癌组织中ＨＰＶ１６ＤＮＡ的检出率分别为２３．０８％、６４．７１％、７１．４３％。而ＨＣＭＶＤＮＡ的检出率则依次为３．８５％、２３．５４％、及４２．８６％，两者检出率呈直线相关（ｒ＝０．９２３）。因此，推测ＨＣＭＶ很可能与ＨＰＶ１６协同作用导致宫颈癌的发生。     

外阴癌的HPV检测及预后的关系

目的研究人乳头瘤病毒（ＨＰＶ）感染与外阴癌的发病和预后的关系。方法将外阴癌病理存档蜡块切片脱蜡，以ＰＣＲ方法扩增其中可能存在的ＨＰＶ－ＤＮＡ（６／１１型和１６／１８型），经琼脂糖凝胶电泳，置紫外光反射仪下观察结果。结果在４９例外阴癌原发灶中，ＨＰＶ－ＤＮＡ１６／１８型的检出率为２６．５％，６／１１型的检出率为２．０％；ＨＰＶ－ＤＮＡ１６／１８型阳性外阴癌的术后生存率好于ＨＰＶ－ＤＮＡ１６／１８型阴性外阴癌。结论一部分外阴癌的发病与ＨＰＶ感染有关，这一部分外阴癌的预后较好；另一部分外阴癌的发病与ＨＰＶ感染无关，但其预后较差。     

PCR检测宫颈病变组织和配偶精液及尿液HPV16、18型的研究

应用聚合酶链反应（ＰＣＲ）技术对３３例女性宫颈病变组织及其配偶精液、尿液进行人乳头瘤病毒（ＨＰＶ）１６、１８型检测。结果：宫颈病变组织、精液和尿液ＨＰＶ１６型检出率分别为５７．６％、５７．６％和１８．１％；ＨＰＶ１８型检出率分别为３０．３％、１２．１％和３．０％。配偶间精液与宫颈病变组织ＨＰＶ１６同型检出率为７３．６％；ＨＰＶ１８同型检出率为３０．０％。研究表明：女性感染ＨＰＶ的同时，配偶也有潜伏感染。实验证实了ＨＰＶ通过精液传播的可能性。     

双温聚合酶链反应检测喉乳头瘤中HPV—DNA

应用双温聚合酶链反应（ＰＣＲ）检测喉乳头瘤石蜡包埋组织中的人乳头瘤病毒（ＨＰＶ）ＤＮＡ》结果与常规ＰＣＲ（三温循环）相同；３０例喉乳头瘤中ＨＰＶ总检出率为４６．７％（１４／３０），ＨＰＶ－６／１１／１６的检出率分别为４０％（１２／３０）、１６．７％（５／３０）和６．７％（２／３０）１６．７％（５／３０）和６．７％（２／３０），ＨＰＶ－６的检出率极显著高于ＨＰＶ－１１／６（ｘ＾２＝１０．６，Ｐ〈）．     

乳头瘤病毒及巨细胞病毒在宫颈癌组织中的协同作用

应用ＤＮＡ－ＤＮＡ斑点杂交技术，对６４例宫颈标本（包括正常宫颈２６例，重度宫颈糜烂１７例，宫颈癌２１例）分别检测ＨＰＶ－１６ＤＮＡ及ＨＣＭＶＤＮＡ。结果发现正常宫颈、宫颈糜烂及宫颈癌组织中ＨＰＶ１６ＤＮＡ的检出率分别为２３．０８％、６４．７１５、７１．４３％。而ＨＣＭＶＤＮＡ的检出率则依次为３．８５％、２３．５４％、及４２．８６％，两者检出率呈直线相关（ｒ＝０．９２３）。因此，推测ＨＣＭＶ很可能与     

HPV的快速检测与分型

以通用引物PCR初筛158例标本，凡HPV感染阳性标本再用型持异引物PCR作分型检测，结果：疣、乳头瘤等良性病变的HPV检出率为61．1％（55／90），主要检出HPV6和／或HPV11，占阳性例数的89．1％（49／55）；食管癌组织有较高的HPV感染率（66．2％，45／68），以检出HPV6，11，16为主，并明显高于HPV8（X2检验，P＜0．01），提示HPV感染可能与本地区食管癌发生密切用关。本研究建立的HPV快速检测和分型方法，用于临床检测和回顾性研究均可获得满意结果。     

人乳头瘤病毒与食管鳞癌及不典型增生相关性的研究

目的研究重庆地区食管癌组织中人类乳头状瘤病毒（HPV）感染与食管鳞癌发生的关系。方法收集112例食管鳞状细胞癌标本和38例非典型增生食管上皮及74例正常食管黏膜组织。运用荧光定量PCR方法分别检测各组织标本中HPV-16、18型的感染状况。结果食管鳞癌组织、非典型增生上皮及正常黏膜中HPV-16DNA的检出率分别为38．4％（43／112）、15．9％（6／38）和5．4％（4／74）。鳞癌组织中HPV-16的检出率明显高于非典型增生上皮和正常食管黏膜（P〈0．05），非典型增生上皮中HPV-16的检出率虽远高于正常食管黏膜，但统计学差异未见显著性；食管鳞癌中HPV-16的阳性检出率与其分化程度未见明显相关。所有组织标本均未检出HPV-18DNA。结论HPV-16的感染可能是重庆地区食管鳞癌发生的之一高危因素；积极开展防治高危型HPV感染对降低该地区食管鳞癌的发病率具有重要的意义。     

以HPV基因通用引物行PCR检测阴茎肿瘤组织中HPV.DNA

为了探讨人乳头瘤病毒感染与阴茎肿瘤的关系。用人乳头瘤病毒基因通用引物行聚合酶链反应,检测５２例阴茎鳞癌。ＨＰＶ阳性率为６７％,３１例为ＨＰＶ－１６,３例为ＨＰＶ１６／１８,１例为ＨＰＶ１１／１８。４例阴茎淋巴结转移灶标本,３例检出与原发癌一致的ＨＰＶ．ＤＮＡ。６例阴茎乳头状瘤标本,４例检出ＨＰＶ－６或１１。２０例正常阴茎包皮均阴性。提示ＨＰＶ有阴茎肿瘤的发生发展中有一定的作用,但与肿瘤的病理分级无     

高危型HPV感染与食管鳞癌组织中HLA-G表达的相关性研究

目的研究人乳头瘤状病毒(HPV)在新乡地区食管鳞癌、癌旁组织中的感染情况和人类白细胞抗原系统G(HLA-G)表达的关系。方法采用基因芯片技术检测114例食管鳞癌组织及癌旁食管正常组织中HPV亚型感染情况,免疫组化SP法检测HLA-G的表达。结果 114例食管鳞癌组织及癌旁食管正常组织中HPV感染阳性率分别为63.2%和6.0%,HLA-G的检出率分别为51.8%、0%,差异均有统计学意义(P<0.05);食管鳞癌存在HPV16、18和52三种亚型感染,其中HPV16感染率最高,HPV52感染率最低。结论新乡地区食管鳞癌组织以HPV16、18和52亚型感染为主;HPV感染和HLA-G蛋白表达与食管鳞癌分化程度正相关;HPV感染食管鳞癌组织与HLA-G蛋白表达存在一定的相关性,可能提示与食管鳞癌发生有较密切关系。     

低密度基因芯片技术检测人乳头瘤病毒基因型

 [目的]利用低密度基因芯片方法对人乳头瘤病毒（HPV）进行分型检测,建立一种经济实用的临床HPV感染的基因型检测方法.[方法]利用低密度基因芯片方法对145例阴道镜门诊病人的宫颈拭子标本进行HPV-DNA检测并分型.[结果]被检人群中共检测出57例HPV阳性,88例HPV阴性,HPV的感染率为39.3%,阳性标本中检测出12种高危型别,2种低危型别.其中单型感染51例（89%）,混合感染6例（11%）,16,31,18,58型检出率较高.[结论]低密度基因芯片是一种快速、简便、特异性强、灵敏度高的检测方法,在HPV和宫颈癌的筛查与诊断中有很大的应用价值.     

某市食管癌组织中人类乳头状瘤病毒感染情况的研究

目的研究河南省食管癌高发区食管癌组织中人类乳头状瘤病毒感染与食管鳞状细胞癌发生的关系。方法利用PCR和免疫组织化学技术检测68例食管癌,15例非典型增生食管上皮和53例正常食管黏膜组织中人类乳头状瘤病毒HPV16／18感染和HPV16／18E6蛋白的表达情况。结果68例食管癌中,HPV DNA阳性率为67．6％（46／68）。在正常食管黏膜、非典型增生上皮及鳞状细胞癌组织中HPV16／18E6的阳性率分别为9．4％（5／53）、40．0％（6／15）和61．2％（42／68）,癌组织中HPV16／18E6蛋白的阳性表达率明显高于非典型增生上皮和正常食管黏膜（P〈0．05）,非典型增生上皮中HPV16／18E6蛋白的阳性表达率亦明显高于正常食管黏膜（P〈0．05）。结论HPV16／18型感染可能在某食管癌高发区食管癌的发生中起重要作用。     

Comparison of polymerase chain reaction and catalyzed signal amplification in situ hybridization methods for human papillomavirus detection in paraffin-embedded cervical preneoplastic and neoplastic lesions

BACKGROUND: Two molecular methods for HPV genotyping in formalin-fixed, paraffin-embedded tissue were evaluated: in house polymerase chain reaction assay (PCR) with consensus and type-specific primers and a novel procedure of in situ hybridization-a catalyzed signal amplification system (CSA-ISH, Genpoint, DAKO, Glostrup, Denmark). The number of HPV positive cases and detected viral types were compared in cervical biopsies and cone specimens according to histopathological diagnosis. Primer efficiency in detecting various types of HPV by PCR method was evaluated. METHODS: DNA samples (101) were used as a template to amplify with three pairs of consensus (MY09/11, GP5+/6 +, CPI/IIG) and four type-specific HPV primers (HPV-6/11, 18, 16 and 33). The according histological tissue sections were analyzed with CSA-ISH method, using commercial HPV biotinylated probes HPV-6/11, 16/18 and 31/33/51. RESULTS: The degree of concordance for PCR and CSA-ISH was 64.4%. In 63 of 101 samples (62.4%), HPV was detected by PCR, while only 35 (34.7%) were positive using CSA-ISH. CSA-ISH found lower percentages for all HPV types, except HPV-6/11. A lower percentage of positive results in all high-grade lesions was detected by CSA-ISH. Multiple infections were detected by PCR in only one sample and in three samples by CSA-ISH. Detection with My09/11 primers followed by Gp5+/6+ primers, in nested reaction, gave the highest number of positive results: 58 of 63 (92%). None of the samples diagnosed as condylomata planum or CIN I was positive for HPV-6/11 (low risk type), which was detected exclusively in condylomata acuminatum group. CONCLUSIONS: A significantly higher number of positive samples was detected with PCR than with CSA-ISH method. CSA-ISH method should be improved, especially in detecting HPV in high-grade lesions. CSA-ISH may be more accurate in detection of multiple infections. GP5+/6+ in nested reaction after MY09/11 detected the highest number of positive results. Samples diagnosed as benign lesions positive on HPV-X must be monitored as possible candidates for progression. CIN I lesions, which were HPV negative, probably will not progress. This finding may be important in planning therapy and avoiding unnecessary treatment.     

Histological and cytological evidence of viral infection and human papillomavirus type 16 DNA sequences in cervical intraepithelial neoplasia and normal tissue in the west of Scotland: evaluation of treatment policy

Biopsy samples from 27 patients referred to a colposcopy clinic in Glasgow for cervical abnormalities were assessed for the relations among colposcopic appearances, cytological and histological diagnosis, expression of papillomavirus antigen, and the presence of human papillomavirus (HPV) types 6, 11, 16, and 18 deoxyribonucleic acid (DNA) sequences. Specimens were from colposcopically abnormal areas of the transformation zone and from colposcopically apparently normal areas of the zone in the same patients (paired matched internal control tissue). All 27 women referred for abnormal smears had colposcopic abnormalities. HPV-16 or 18 DNA sequences were detected in 20 of the 27 colposcopically abnormal biopsy samples and 13 of the 27 paired normal samples. Twelve samples of colposcopically normal tissue contained histological evidence of viral infection but only four of these contained HPV DNA sequences. The other nine samples of colposcopically normal tissue which contained HPV DNA sequences were, however, histologically apparently normal. HPV-6 and 11 were not detected. Integration of the HPV-16 genome into the host chromosome was indicated in both cervical intraepithelial neoplasia and control tissues. In two thirds of the HPV DNA positive samples the histological grade was classed as normal, viral atypia, or cervical intraepithelial neoplasia grade 1. Papillomavirus antigen was detected in only six of the abnormal and three of the normal biopsy samples, and HPV DNA was detected in all of these. The detection of HPV DNA correlates well with a combination of histological and cytological evidence of viral infection (20 of 22 cases in this series). A poor correlation between the site on the cervix of histologically confirmed colposcopic abnormality and the presence of HPV DNA sequences implies that a cofactor other than HPV is required for preneoplastic disease to develop. A separate study in two further sets of biopsy samples examined the state of HPV DNA alone. The sets were (a) 43 samples from cervical intraepithelial neoplasia and nine external controls and (b) 155 samples from cervical intraepithelial neoplasia, cervical cancer, vulval intraepithelial neoplasia, and vulval cancer and external controls. HPV-11 was found in only two (4.7%) of the 43 specimens from cervical intraepithelial neoplasia, whereas HPV-16 was found in 90 (58%) of the other 155 specimens. These results also suggest that HPV subtype is subject to geographical location rather than being an indicator of severity of the lesion or of prognosis.     

DNA sequences of human papillomavirus types 11, 16, and 18 in lesions of the uterine cervix in the west of Scotland

Punch biopsy specimens of the cervix were examined both histologically and for the presence of human papillomavirus (HPV) DNA sequences. The presence of HPV DNA sequences was sought with the Southern blot technique using radioactively labelled HPV-6, 11, 16, and 18 DNA probes, both together and separately. Twenty six biopsy specimens were examined. Histological examination showed cervical intraepithelial neoplasia grade 2 or 3 in 16 specimens, viral changes (koilocytosis) in four, and inflammation or a normal appearance in three. Eleven specimens were negative for HPV DNA sequences, 10 contained HPV-16 DNA, four contained HPV-18 DNA, and one contained both HPV-18 and HPV-11 DNA. Episomal HPV-16 DNA was detected in one case of cervical intraepithelial neoplasia grade 3 and in five cases of cervical intraepithelial neoplasia grade 2/3 with koilocytosis; and episomal HPV-18 DNA was found in two specimens classed as cervical intraepithelial neoplasia grade 2/3, one of which also contained HPV-11 DNA, and in one specimen that showed viral changes alone. Integrated HPV DNA was found in six specimens (four with HPV-16 DNA and two with HPV-18 DNA), including two cases of chronically inflamed cervix with no histological evidence of viral infection or cervical intraepithelial neoplasia. Detection of viral DNA in early lesions may identify patients at risk of malignant progression. This is the first report of HPV-18 DNA in cervical intraepithelial neoplasia in Scotland.