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〈）．     

小儿咽喉乳头状瘤组织人乳头瘤病毒检测及型别分析

目的：探讨人乳头瘤病毒感染和小儿咽喉乳头状瘤的关系。方法：应用ＰＣＲ和ＰＣＲ产物斑点杂交技术检测３５例小儿咽喉乳头瘤组织和１０例小儿声带小结（对照组）ＨＰＶ６、１１、１６、１８、３３五个型别的ＤＮＡ。结果：乳头瘤组织ＨＰＶ感染率为９１．４％（３２／３５）,其中ＨＰＶ６型检出率为５４．２％（１９／３５）,ＨＰＶ１１型感染率为２５．７％（９．３５）,多重型ＨＰＶ６＋１１感染率为１１．４％（４／３５）;     

子宫颈组织人乳头瘤病毒感染状况的研究

应用兼并复合人乳头瘤病毒－聚合酶链反应（ＨＰＶ－ＰＣＲ）技术，对７４例外观正常的子宫颈分泌物拭子标本进行人乳尖瘤病毒（ＨＰＶ）ＤＮＡ的测定，结果：正常子宫颈ＨＰＶＤＮＡ检出率为３２．４３％（２４／７４），其中ＨＰＶ６、１１型为６．７６％（５／７４），ＨＰＶ１６、１８型为４．０５％（３／７４），ＨＰＶ其它型为２１．６２％（１６／７４），外观正常的子宫颈ＨＰＶ感染的主要易感年龄为２６～３４岁的青年妇女     

喉鳞癌及乳头状瘤组织中人乳头瘤病毒DNA的PCR检测

①目的探讨人乳头瘤病毒１６型和１８型（ＨＰＶ１６和ＨＰＶ１８）与喉鳞癌、喉乳头状瘤的关系。②方法应用聚合酶链反应（ＰＣＲ）技术检测３４例喉乳头状瘤、８３例喉鳞状细胞癌石蜡包埋组织中ＨＰＶ１６及ＨＰＶ１８的ＤＮＡ．③结果喉乳头状瘤及喉鳞癌组织中ＨＰＶ１６的检出率为１４．７１％和３０．１２％，两组比较差异无显著性（χ２＝３．００６，Ｐ＞０．０５）；ＨＰＶ１８在喉乳头状瘤组织中未检出，而在喉鳞癌组织中为６．０２％，两组比较，差异无显著意义（χ２＝０．０８７，Ｐ＞０．０５）。高分化喉鳞癌组织中ＨＰＶ１６的检出率显著高于中分化鳞癌（χ２＝６．０３２，Ｐ＜０．０５），而高分化与低分化、中分化与低分化鳞癌之间的差异均无显著性（χ２＝０．３５１，１．５００，Ｐ均＞０．０５）；不同病理分化喉鳞癌组织中ＨＰＶ１８的检出率差异无显著性（χ２＝０．０６０，Ｐ＞０．０５）。④结论喉鳞癌的发生与ＨＰＶ感染密切相关，尤以ＨＰＶ１６明显     

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，喉其他病变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。     

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。     

子宫颈组织人乳头瘤病毒感染状况的研究

应用兼并复合人乳头瘤病毒—聚合酶链反应（ＨＰＶ－ＰＣＲ）技术，对７４例外观正常的子宫颈分泌物拭子标本进行人乳头瘤病毒（ＨＰＶ）ＤＮＡ的测定。结果：正常子宫颈ＨＰＶＤＮＡ检出率为３２．４３％（２４／７４），其中ＨＰＶ６、１１型为６．７６％（５／７４）；ＨＰＶ１６、１８型为４．０５％（３／７４）；ＨＰＶ其它型为２１．６２％（１６／７４）。外观正常的子宫颈，ＨＰＶ感染的主要易感年龄为２６～３４岁的青年妇女。本研究对女性生殖道ＨＰＶ感染的状况及传播途径进行了探讨。     

用多聚酶链反应探讨宫颈人乳头瘤病毒感染

目的了解温州地区ＨＰＶ感染与宫颈炎的关系，为防治该病提供依据。方法用多聚酶链反应（ＰＣＲ）检测１２８例慢性宫颈炎及４８例正常宫颈标本组织中的人乳头瘤病毒（ＨＰＶ）ＤＮＡ，并对ＨＰＶ进行１６、１８型别分析。结果慢性宫颈炎组ＨＰＶ检出率为３２．９％（４２／１２８），ＨＰＶ１６、１８及双重感染（１６＋１８）各占其中的５４％（２３／４８）、１１．９％（５／２４）、１１．９％（５／２４），对照组．ＨＰＶ的检出率为６．３％（３／４８）。两组ＨＰＶ阳性检出率有显著性差异（Ｐ＜０．０５）。结论ＨＰＶ感染，尤其ＨＰＶＩ６感染与慢性宫颈炎相关，但与年龄大小无关。     

应用PCR技术对女性生殖道尖锐湿疣及其它良性病变HPV—DNA…

本研究应用ＰＣＲ技术对女性生殖道尖锐湿疣、可疑湿疣、假性湿疣、宫颈息肉、宫颈糜烂、慢性阴道炎等良性病变以及正常阴道组织进行ＨＰＶ－ＤＮＡ检测，结果ＨＰＶ－ＤＮＡ阳性检出率分别为９８．２％（５６／５７）、７１．４％（１５／２１）、４０．４％（１０／４７）、７６．２％（１６／２１）、７７．３％（１７／２２）、１６％（４／２５）和１０．７％（３／２８），ＨＰＶ６和１１型感染率分别占８５．７％（４８／５６     

用PCR检测新疆部分女性尖锐湿疣与宫颈上皮内瘤变组织HPV的…

应用多重引物人乳头瘤病毒（ＨＰＶ）６ｂ、１１、１６、１８型聚合酶链反应（ＰＣＲ）技术检测７４例新疆南疆、北疆部分地区女性生殖道尖湿疣（ＣＡ）与宫颈上皮内瘤变（ＣＩＮ）组织中人乳头瘤病毒（ＤＮＡ（ＨＰＶ ＤＮＡ）。其中在南疆检测出ＣＡ１７例，ＣＩＮⅡ级２０例，在北疆检测出ＣＡ２２例，ＣＩＮⅠ－Ⅱ级２例，ＣＩＮⅢ级１３例。结果：总ＨＰＶＤＮＡ检出率 ９４．１％、９０．０％、９０．９％、８６．７％。ＣＩ     

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。     

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.     

食管癌组织中人乳头瘤病毒的PCR检测

应用ＰＣＲ对汕头地区６０例食管癌石蜡包埋标本进行人乳砂瘤病毒ＤＮＡ序列检测。结果：ＨＰＶ－ＤＮＡ总检出率为６３．３％，ＨＰＶ－６、１１、１６、１８型的检出率分别为３０．０％，４０．０％，３０．０％和１０．０％，其中多重感染占阳性例数的６０．５％。初步结果表明，汕头地区食管癌组织中有较高的ＨＰＶ感染率，此与食管癌的发生可能有密切关系。     

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     

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。     

A high throughout assay for human papillomavirus genotypes with fluorescence polarization

Objective To develop a simple, cheap, quick, accurate and practical method for a high throughout genotypes assay of human papillomavirus (HPV) DNA.Methods Crude DNA was extracted by a simplified proteinase K digesting method. HPV common conservative primers: GP5 /6 system was used to amplify HPV DNA in 127 samples of condylomata acuminatum (CA) and cervical scrapes by PCR, then the PCR product was assayed using a template directing terminator incorporation (TDI) and genotypes were detected with fluorescence polarization (FP). Major HPVs type-specific probes (HPV6, 11, 16, 18, 31, 33, 35 and 58) designed by us were hybridized with the specific PCR products and a special fluorescent ddNTP terminator was directly added to the end of the probe under direction of specific PCR products.The results were measured with FP and compared with the results of the DNA sequence.Results Compared with the results of DNA sequencing, the results detected with fluorescence polarization were all correct. The proposed method could detect more than one type of HPV infection,but DNA sequencing method could not. The positive rate of HPV was 100% in 78 CA biopsies.Among them, there were 14 HPV double infections [HPV6B and 11(9 cases), HPV11 and 16(4),HPV11 and 18(1)], 5 HPV triple infections [HPV6B,11 and 16(4), HPV11,16 and 18(1)], and one HPV quadruple infection (HPV6B, 11,16 and 18). The positive rate of HPV was 77% in the 49cervical scrapes. Six HPV double infections [HPV6B and 11(2), HPV11 and 16(1), HPV6B and 16(1), HPV16 and 18(1), HPV18 and 58 (1)], 3 HPV triple infections [HPV6B, 11 and 16(2),HPV11,16 and 18(1)] and one HPV quadruple infection (HPV6B,11,16 and 18) were detected incervical cancer scrapes.Conclusions The proposed method allowed a high throughout, special, simple, rapid, automatic and economical detection of HPV-DNA genotyping without a use of labeling probes. It can detect multiple HPV genotype infection and will be and useful tool in HPV genotype screening.     

宫颈癌患者血液中人乳头瘤病毒检出率的临床意义

目的:探讨宫颈癌患者血液中人乳头瘤病毒(HPV)DNA检出率对宫颈癌的诊断及病情进展评价的临床意义。方法:应用巢式PCR检测48例宫颈癌,55例宫颈高度病变(HG),6例宫颈低度病变(LG),29例宫颈炎症患者血浆中HPV-16,18DNA。分析各组患者血中HPVDNA检出率的差异,以及检出率与宫颈癌临床分期的相关性。结果:血浆中HPVDNA在宫颈癌组、HG组、LG组及宫颈炎症组中的检出率分别为56.3%、41.8%、16.7%和37.9%,其中HPV-16分别为20.8%、5.5%、0.0%和6.9%;HPV-18分别为50.0%、38.2%、16.7%和31.0%。HPV-16和HPV-18均为阳性者8例,其中宫颈癌Ⅰ期3例,Ⅱ期4例,CINⅢ1例。在9例有淋巴结转移的患者中,6例血浆HPVDNA阳性。血浆HPVDNA在宫颈癌组中检出率高于其他3组,但无统计学差异。结论:宫颈癌患者血中HPV检出率高于CIN和宫颈炎症患者,并且在发生转移的宫颈癌患者中检出率更高,但血浆HPVDNA检出率与宫颈癌临床分期无关。宫颈癌患者血中HPV双重感染率较高。     

新疆不同民族喉乳头状瘤与喉癌组织中不同类型HPV的检测

目的：探讨新疆部分民汉族喉乳头状瘤（LP）和喉癌（LC）与人乳头状瘤病毒（HPV）感染的关系。方法：应用聚合酶链反应的（PCR）技术，对83例喉癌患者（维吾尔族27例，哈萨克族4例，汉族52例），63例喉乳头状瘤患者（维吾尔族33例，汉族30例），20例声带炎性息肉（LIP）患者（维、汉各10名），总共166例分别进行HPV6／11的9．6％（8／83）和LIP的0．0％（0／20）（P＜0．05）；（2）LC组织中HPV16／18感染的阳性率为37．3％（31／83）明显高于LP的6．3％（4／63）和LIP的0．0％（0／20（P＜0．05）；（3）民汉之间的差异无统计学意义（P＞0．05）。结论：（1）民汉LP的发生主要与HPV6／11感染有关；LC主要与HPV16／18感染关系密切，HPV感染可引起细胞基因的异常调控，可能是喉上皮组织发展成为肿瘤病变的促进因子。     

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.     

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.