婴幼儿呼吸道乳头瘤病人乳头瘤病毒的检测

目的：研究人乳头瘤病毒（ＨＰＶ）与婴幼儿呼吸道乳头瘤病临床病理特征之间的关系。方法应聚合酶链式反应－限制性片段长度多态性技术，对１３例婴幼儿呼吸道乳头瘤病组织中的ＨＰＶ进行定型分析。结果：ＨＰＶ阳性率为１００％，其中ＨＰＶ６占６１．５％（８／１３），ＨＰＶ１１占３０．８％（４／１３），ＨＰＶ１５占７．７％（１／１３）。无一例ＨＰＶ１６、３１、３３、５２、５８阳性。每例患者多次手术标本检测结果一致。     

小儿喉乳头状瘤HPV-DNA及体液免疫检测

目的：探讨小儿喉乳头状瘤（ＪＬＰ）人乳头瘤病毒（ＨＰＶ）感染途径及发病机理。方法：采用ＰＣＲ及ＰＣＲ产物斑点杂交技术检测ＪＬＰ组织ＨＰＶ－ＤＮＡ;散射免疫比浊法测定血清Ｉｇ及补体Ｃ３。结果：ＪＬＰ组织ＨＰＶ总感染率为９５％（１９／２０）,其中ＨＰＶ。型为５５％（１１／２０）,ＨＰＶ１１为３０％（６／２０）,ＨＰＶ６＋１１型为１０％（２／２０）;ＪＬＰ患者血清ＩｇＧ、ＩｇＡ、ＩｇＭ、Ｃ３值正常,对照     

上呼吸道乳头状瘤HPVDNA的检测

应用多重引物ＰＣＲ技术对４７例复发性呼吸道乳头状瘤病（ＲＲＰ）和９例声带息肉石蜡包埋组织中ＨＰＶ６／１１、１６、１８ＤＮＡ进行了检测。结果发现：①ＲＲＰ组织中ＨＰＶ６／１１ＤＮＡ的阳性率为６８．１％（３２／４７），其中喉乳头状瘤、口咽乳头状瘤、鼻腔和鼻前庭乳头状瘤组织中ＨＰＶ６／１１ＤＮＡ的阳性率分别为７０．４％（１９／２７）、６６．７％（４／６）和６４．３％（９／１４）；②所有ＲＲＰ标本中均未发现特异性的ＨＰＶ１６和ＨＰＶ１８ＤＮＡ；③９例声带息肉标本中四种型号的ＨＰＶＤＮＡ全部阴性。实验结果表明，ＲＲＰ的发病与ＨＰＶ６／１１感染密切相关。多重引物ＰＣＲ检测ＨＰＶＤＮＡ具有敏感、特异、快速、简便等优点，适合于临床广泛开展和进行流行病学调查。     

鼻腔鼻窦内翻性乳头状瘤人类乳头状瘤病毒DNA检测

为了解鼻腔鼻窦内翻性乳头状瘤与人类乳头状瘤病毒（ＨＰＶ）之间的关系，采用聚合酶链反应，对３８例患者的４４个鼻腔鼻窦内翻性乳头状瘤的病理组织蜡块进行ＨＰＶ－ＤＮＡ的检测。结果显示，３８例中３０例患者的３０个瘤组织蜡块呈ＨＰＶ阳性，总感染率为６８．２％，其中３０例次ＨＰＶ１１型阳性（６８．２％），１８例次ＨＰＶ１６型阳性（４０．９％），２例次ＨＰＶ１８型阳性（４．５％），其中１８例ＨＰＶ１１，１６、２例ＨＰＶ１１，１８呈双重感染。试验表明，ＨＰＶ与鼻腔鼻窦内翻性乳头状瘤在病因上有一定的相关性。     

喉癌人乳头瘤病毒感染和p53蛋白表达的研究

对４４例喉鳞癌和１６例声带息肉患者采用原位核酸杂交技术探测其人乳头瘤病毒（ＨＰＶ）６Ｂ，１１，１６，１８型ＤＮＡ同源序列及ＬＳＡＢ免疫组化法探测其Ｐ５３蛋白的表达，结果：（１）喉癌与ＨＰＶ１６／１８杂交阳性率为４３．２％（１９／４４），声带息肉为１２．５％（２／１６）（Ｐ〈０．０５）（２）喉癌ｐ５３蛋白阳性率为５６．８％（２５／４４），声带息肉全部为阴性；（３）喉癌ＨＰＶ１６／１８杂交及ｐ５３蛋白     

国人喉癌与人类乳头状瘤病毒相关性的研究

本项研究对１２４例喉不同病变的新鲜组织标本，采用共同引物和多重引物ＰＣＲ的方法，进行ＨＰＶＤＮＡ检测。用共同引物ＰＣＲ检测ＨＰＶ＿（６，１１，１６，１８，３１，３３，３５，４２，５８）九个型别的感染，阳性病例再用多重引物ＰＣＲ进一步分型。结果，①喉癌组：ＨＰＶ感染的总阳性率为４９．１％，其中ＨＰＶ＿（１８）型阳性率１５．８％，ＨＰＶ＿（１６）型为１２．３％，ＨＰＶ＿（１６）和＿（１８）双重型感染为５．３％，ＨＰＶ＿（６／１１）和＿（１８）型混合感染为３．５％，其它型为１２．３％。②颈转移淋巴结组：总阳性率为２１．４％，其中ＨＰＶ＿（１６）、ＨＰＶ＿（１８）及ＨＰＶ＿（１６）和＿（１８）双重型感染各为７．１％。③癌前病变组：ＨＰＶ感染阳性率为１１．１％，为ＨＰＶ＿（６／１１）和＿（１８）型混合感染。④声带息肉组：阳性率７．１％，为ＨＰＶ＿（６／１１）型感染。⑤癌旁及癌周正常喉组织：均为ＨＰＶＤＮＡ阴性。本文对ＨＰＶ在喉癌中致病作用进行了讨论。     

鼻腔鼻窦内翻性乳头状瘤人类乳头状瘤病毒DNA检测

为了解鼻腔鼻窦内翻性乳头状瘤与人类乳头状瘤病毒（ＨＰＶ）之间的关系，采用聚合酶链反应，对３８例患者的４４个鼻腔鼻窦内翻性乳头状瘤的病理组织蜡块进行ＨＰＶ－ＤＮＡ的检测。结果显示，３８例中３０例患者的３０个瘤组织蜡块呈ＨＰＶ阳性，总感染率为６８．２％，其中３０例次ＨＰＶ１１型阳性（６８．２％），１８例次ＨＰＶ１６型阳性（４０．９％），２例次ＨＰＶ１８型阳性（４．５％），其中１８例ＨＰＶ１１，１６、２     

原位杂交在成年和幼年型喉乳头状瘤HPV病因学的对比研究

探讨成年和幼年型喉乳头状瘤ＨＰＶ感染发病差异及其影响因素。方法：用地高辛配基（Ｄｉｇｏｘｉｇｅｎｉｎ）标记ＨＰＶ６和ＨＰＶ１１型作探针，原位核酸杂交方法在２９例成年型喉乳头状瘤（ＡＬＰ）和２１例幼年型喉乳头状瘤（ＪＬＰ）石蜡包埋标本检测ＨＰＶ同源序列。结果：ＡＬＰＨＰＶ６和ＨＰＶ１１阳性率分别为４１４％（１２／２９）和４８３％（１４／２９）；ＪＬＰＨＰＶ６及ＨＰＶ１１阳性率均为７６２％（１６／２１）。ｘ２统计示：两型喉乳头状瘤ＨＰＶ６及ＨＰＶ１１阳性率明显不同（ＨＰＶ６ｘ２＝５９９，ＨＰＶ１１ｘ２＝３９５，Ｐ均小于００５）。结论：１）ＡＬＰ和ＪＬＰＨＰＶ感染发病存在差异。２）ＡＬＰ除了ＨＰＶ感染外，其促发因素不可忽视，ＪＬＰ更倾向于依赖ＨＰＶ感染而发病。     

国人喉癌与人类乳头状瘤病毒相关性的研究

本项研究对１２４例喉不同病变的新鲜组织标本，采用共同引物和多重引物ＰＣＲ的方法，进行ＨＰＶ ＤＮＡ检测。用共同引物ＰＣＲ检测ＨＰＶ６．，１６，１８，３１，３３，３５，４２，５８九个型的感染，阳性病例再用多重引物ＰＣＲ进一步分型。结果，（１）喉癌组；ＨＰＶ感染的总阳性率为４９．１％，其中ＨＰＶ１８型阳性率１５．８％，ＨＰＶ１６型为１２．３％，ＨＰＶ１６和１８双重感染为５．３％，ＨＰＶ６／１１和１８型     

PCR和PCR产物斑占杂交技术检测成人咽喉病变中人乳头状？…

目的：研究成人咽喉部良、恶性病变与人乳头状瘤病毒（ＨＰＶ）感染的关系。方法：应用聚合酶链反应（ＰＣＲ）和斑点杂交技术,对５５例咽喉不同病变的新鲜组织标本进行ＨＰＶ６,１１,１６,１８,３３共５型ＨＰＶ－ＤＮＡ感染的检测。结果：在咽乳状瘤组ＨＰＶ感染率为６０％（６／１０）,喉乳头状瘤组为７０％（７／１０）,喉鳞状上皮非典型增生组为２０％（１／５）,声带息肉组为２０％（１／５）,喉癌组为２０％（１／５     

婴幼儿咽喉乳头瘤组织人乳头瘤病毒感染的探讨

目的　探讨温州地区人乳头瘤病毒感染和婴幼儿咽喉乳头状瘤的关系。方法　应用聚合酶链反应和核酸斑点杂交技术检测 35例婴幼儿咽喉乳头瘤组织和 10例对照组组织 (小儿声带小结 )HPV6、11、16、18、3 3 5个型别的DNA。结果　乳头瘤组织HPV感染率为 91 4% (30 / 35 ) ,其中HPV6型检出率为 5 4 2 % (19/ 35 ) ,HPV11型感染率为 2 5 7% (9/ 35 ) ,多重型别HPV6 11感染率为 11 4% (4/ 35 ) ;HPV16、18、3 3 型 ,均为阴性。对照组各型检测结果均为阴性 ,两者对比差异有高度显著性 (P <0 0 0 1)。结论　温州地区婴幼儿咽喉乳头瘤的发生与HPV感染密切相关 ,尤以HPV6感染为主。PCR结合核酸斑点杂交技术检测HPV具有敏感性高、特异性强的优点 ,值得推广。     

HPV11对小儿喉乳头状瘤预后的影响

目的 :研究人乳头状瘤病毒 (HPV)型别对小儿喉乳头状瘤 (JLP)预后的影响。方法 :应用聚合酶链反应结合斑点杂交技术对 2 5例JLP的石蜡标本进行HPV定型分析 ,并统计HPV11、HPV6 感染组的气管切开率和术后复发率。结果 :HPV总检出率为 96.0 % ,其中HPV11为 5 6.0 % ,HPV6 为 4 0 .0 % ,HPV16、18、33无一例阳性。HPV11感染组的气管切开率为 71.4 % ,术后复发率为 85 .7% ;HPV6 感染组的气管切开率为 3 0 .0 % ,术后复发率为4 0 .0 %。两组分别比较 ,其差异均有显著性意义 (P <0 .0 5 )。结论 :HPV6、11与JLP发生密切相关 ,HPV11感染与JLP的喉梗阻和术后复发率相关 ,HPV11感染可作为JLP预后评判的重要依据。     

Detection of human papillomavirus genome in nasolaryngeal papillomas using digoxigenin labeled DNA probes

It is being reported that human papillomavirus (HPV) has been implicated in the pathogenesis of various neoplastic lesions of the genital organs. To investigate the etiological role of HPV and its types in nasolaryngeal papillomas, we retrospectively analyzed HPV genomes by nucleic acid hybridization methods; for detecting DNA and mRNA, we employed the recently developed nonradioactive (digoxigenin labeled) DNA probes and compared the results by radioisotope methods. In total, 43 cases of papillomatous lesions were examined. They were verruca vulgaris of the nasal vestibule (Nr = 2), nasal inverted papilloma (IP, Nr = 26), and laryngeal papilloma (Nr = 15). HPV types examined were type 2, 6, 11, 16 and 18. Two cases of verruca vulgaris were shown to contain HPV-2 DNA and its mRNA by in situ hybridization. HPV-11 DNA was detected in 3 cases (12%) of nasal inverted papilloma whereas HPV-16 was detected in 1 case (4%); the latter case was associated with squamous cell carcinoma. These results suggest that HPV may be implicated in the development of IP, and HPV-16 may play an important role in the malignant transformation of IP. In the cases of multiple laryngeal papilloma (Nr = 8, one juvenile type and 7 adult type), either HPV-6 or HPV-11 was detected at the high rate (6/8, 75%). The presence of the HPV genomes provides strong evidence for the HPV etiology of these laryngeal papillomas. Whereas in the cases of adult single laryngeal papilloma (Nr = 7), HPV was not detected. Technically, the sensitivity of digoxigenin (DIG) labeled DNA probe was almost same as 35S labeled probe by dot blot hybridization, thus we applied DIG labeled probe to Southern blot hybridization with low background. By in situ hybridization using digoxigenin labeled probes, the rates of HPV detection were almost equal to those by 35S labeled probes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection and typing of papillomavirus DNA in formaldehyde-fixed paraffin-embedded tissue

Clinical specimens from nine patients with papillomatosis of the vocal cords and three patients with vocal cord polyps were evaluated for the presence of human papillomavirus (HPV) DNA using two complementary molecular hybridization techniques. In one method, involving polymerase chain reaction (PCR) amplification, HPV DNA sequences were replicated in vitro from tissue DNA extracted from paraffin sections prior to hybridization. Polymerase chain reaction amplification was compared with the standard method of Southern blot hybridization. Results of the two techniques for all nine laryngeal papillomas agreed completely: five patients harbored HPV type 6 and four HPV type 11. Both PCR amplification and Southern blot hybridization found two of the three polyps to be free of HPV infection, while PCR detected HPV type 18 in one polyp specimen that was reported negative by Southern blot hybridization, suggesting a greater sensitivity of PCR. Our results demonstrate that PCR amplification is as reliable and at least as sensitive as Southern blot hybridization. Moreover the PCR technique opens the way to the undertaking of a whole variety of retrospective studies using formaldehyde-fixed paraffin-embedded tissues.     

Human papilloma virus infection and expression of p16 protein in laryngeal papilloma and laryngeal carcinoma]

OBJECTIVE: To evaluate the role of human papilloma virus (HPV) infection and inactivation of p16 gene in laryngeal papilloma (LP) and laryngeal squamous cell carcinoma (LC). METHODS: HPV consensus primers direct in situ polymerase chain reaction (ISPCR) and immunohistochemical method were applied to detect the presence of HPV genomes (1, 6, 8, 11, 13, 16, 18, 30, 31, 32, 33, 45, 51) and the expression of p16 protein respectively in 93 cases of formalin-fixed, paraffin-imbedded specimens, which contained 46 cases of LPs [adult-onset laryngeal papilloma (ALP) 21, juvenile-onset laryngeal papilloma (JLP)25], 26 cases of LCs, 6 cases of normal tissues adjacent to carcinoma, and 15 cases of vocal noduli. RESULTS: (1) The difference of positive rates of HPV-DNA in JLP group (84%, 21/25) and other groups were statistically significant (chi 2 test, P < 0.05). The difference of positive rates of HPV-DNA in ALPs(38.1%, 8/21), in LCs(19.2%, 5/26), in vocal noduli(0%, 0/15), and in normal tissues adjacent to carcinoma(0%, 0/6) were not significant statistically (chi 2 test or Fisher's exact probability test, P > 0.05). (2) The positive rates of expression of p16 protein in ALP group(57.1%, 12/21) and LC group(38.5%, 10/26) were significantly lower than that in vocal nodule group(93.3%, 14/15), in JLP group(88%, 22/25), and in normal tissues adjacent to carcinoma group (100%, 6/6) (chi 2 test or Fisher's exact probability test, P > 0.05). There were no significant differences of positive rates of expression of p16 protein between ALP group and LC group, and between JLP group and vocal nodule group (chi 2 test, P > 0.05). (3) In LPs, the difference of positive rates of p16 protein expression between HPV positive cases and HPV negative cases was significant statistically (chi 2 test, P < 0.05). In LCs, there was no difference in p16 protein expression rate between the two teams(Fisher exact probability test, P > 0.05). CONCLUSION: The pathogenesis of JLP is closely associated with HPV infection and not associated with the inactivation of p16 gene. Conversely, the pathogenesis of ALP and LC is associated with the inactivation of p16 gene and not associated with the HPV infection.     

新疆部分地区儿童复发性喉乳头状瘤HPV6和HPV11病毒感染在维吾尔族及汉族患者之间的差异

目的：研究新疆部分地区喉乳头状瘤病毒HPV6,HPV11在汉族及维吾尔族儿童复发性喉乳头状瘤（JRRP）患儿中的表达差异。方法：采用聚合酶链反应（PCR）技术对喉乳头状瘤组织中HPV11和HPV6 DNA进行定型分析,结合回顾性分析1996—01—2008—03期间在新疆医科大学第一附属医院耳鼻咽喉科连续收治的42例JRRP患者。结果：HPV6／11阳性检出率97．61％（41／42）,HPV6阳性36．58％（41／15）,HPV11阳性63．41％（41／26）,HPV6阳性组中维族HPV6阳性53．33％（8／15）,汉族HPV6阳性46．67％（7／15）,HPV11阳性组中维族HPVll阳性65．38％（17／26）,汉族HPV11阳性34．61％（9／26）。结论：新疆部分地区JRRP以HPV11,6感染为主,HPV11感染者占多数,HPV病毒类型与维汉间的发病率之间差异无统计学意义。     

Human papillomavirus in laryngeal papillomas and in adjacent normal epithelium

Eleven adults with laryngeal papillomas were studied for the presence of human papillomavirus (HPV) DNA by in situ hybridization. As well as from the papillomas, three additional biopsies were taken from the normal-appearing mucosa as follows: the involved vocal cord, the opposite vocal cord (when the papilloma was unilateral), and from the ventricular fold on the side of the lesion. These normal tissues were analysed by polymerase chain reaction (PCR) to detect HPV DNA. All except one of the 11 papillomas contained HPV DNA; nine were HPV 6/11 DNA positive and one positive for HPV 16 DNA. The normal-appearing laryngeal mucosa harboured HPV DNA in eight out of 11 patients. The present results strongly support the concept that the adult-type laryngeal papilloma is an HPV-induced lesion, mostly due to HPV types 6 and 11. The persistence of HPV DNA in the adjacent normal epithelium is consistent with the frequent recurrence of these lesions.     

人咽喉部良恶性肿瘤与乳头状瘤病毒关系的研究

采用免疫组化及ＤＮＡ斑点杂交技术检测人咽喉部乳头状瘤及鳞状细胞癌组织中人乳头状瘤病毒（ＨＰＶ）壳蛋白抗原及ＨＰＶ６、１１、１６、１８型ＤＮＡ。１１例乳头状瘤ＨＰＶ抗原与ＨＰＶ ＤＮＡ阳性率均为４５．５％。２２例鳞状细胞癌ＨＰＶ抗原阳性率２２．７％，ＨＰＶ ＤＮＡ阳性率２７．３％。乳头状瘤ＨＰＶ检出率与组织学检查的结果相符。提示咽喉部乳头状瘤及鳞状细胞癌的发生、发展与ＨＰＶ感染有关。