DIFFERENTIATION OF PSEUDOCONDYLOMA OF VULVA AND CONDYLOMA ACUMINATA BY DOT BLOT HYBRIDIZATION AND POLYMERASE CHAIN REACTION

This study differentiated pseudocondyloma of vulva from condyloma acuminata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases of pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the study. The genital lesions were examined clinically and were biopsied. Each biopsy was subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82. 6%) out of 23 cases of condyloma acuminata and 2(25% ) out of 8 cases pseudocondyloma of vulvae (P<0. 05). PCR detected HPV DNA in 51 (92. 7%) out of 55 cases of condyloma acuminata , compared with none in 23 cases of pseudocondyloma (P<0.001 ). HPV DNA was present in the majority of condyloma acuminata specimens. HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful technique for HPV DNA detection.

COMPARISON BETWEEN IMMUNOFLUORESCENCE AND PCR IN DETECTING HUMAN PAPILLOMA VIRUS IN CONDYLOMA ACUMINATA

Objective To compare the effectiveness of immunofluorescence and polymerase chain reaction （PCR） in detecring human papilloma virus （HPV） in condyloma acuminata （CA）. Methods HPVs in CA tissues from 60 patients were detected by immunofluorescence and PCR, respectively. Different subtypes of HPVs were also identified with restriction fragment length polymorphism （RFLP） . Resuits The positive detective rates of immunofluorescence and PCR were 56. 67 % （34/60） and 96. 67 % （58/ 60）, respectively （ P 〈 0. 01 ）. RFLP results showed HPV6 and HPV11 were the main subtypes in the detected virus, which accounted for 98.28%. Conclusion The sensibility of PCR is superior to that of immunofluorescence.

Human papillomavirus DNA and P16～(INK4A) expression in concurrent esophageal and gastric cardia cancers

AIM:To investigate the relationship between human papillomavirus (HPV) infection and concurrent esophagus and gastric cardia cancer from the same patient (CC) and examine the significance of P16 INK4A protein expression.METHODS:Polymerase chain reaction was used to detect the presence of HPV type16 (HPV16).The expression of P16 INK4A protein was detected using immunohistochemistry.RESULTS:Among the CC specimens,HPV16-DNA was found in eight cases of esophageal squamous cell carcinoma (ESCC) and five cases of gastric cardia adenocarcinoma (GCA),respectively (47% vs 29%),and two of both ESCC and GCA.P16 INK4A was highly expressed in both ESCC and GCA.In the HPV-associated positive CC,higher P16 INK4A expression was observed in the GCA than in the ESCC (75% vs 25%,P < 0.05).CONCLUSION:HPV16 as a correlated risk factor may play an important role in the development of ESCC and GCA.P16 INK4A may be a screening index in the HPVassociated carcinoma of gastric cardia.

Comparison of Human Papillomavirus Detection and Genotyping with Four Different Prime Sets by PCR-Sequencing

Objective To assess and compare the Human Papillomavirus（HPV） detection efficiency and the potential clinical utility of PCR sequencing‐based technology.Methods Four HPV consensus primer sets（GP5＋/6＋,MGP,MY09/11,and PGMY09/11） were used in order to amplify a broad spectrum of HPV types for HPV infection in 325 cervical samples and the PCR products were sequenced afterwards for the HPV genotyping.Results The HPV‐positive rate was 75.4%,of which 35.5% harbored more than one HPV genotype.A total of 36 different genotypes was found,with HPV 16（24.1%） being the most prevalent,followed by HPV 58（13.3%） and HPV 52（9.6%）.There were substantial to almost perfect agreements between different primer sets regarding HPV detection efficiency,with the kappa value varying from 0.751 to 0.925,MGP,and PGMY09/11 were the most effective in detecting multiple infections（P0.001）.With each of the primer sets,a board range of HPV types could be identified,though there were several differences for a few genotypes.Conclusion The substantial agreement between PCR‐sequencing and HC2 for the detection of high‐risk HPV（kappa=0.761） indicated that PCR‐sequencing is also suitable for routine HPV screening.

Urine versus brushed samples in human papillomavirus screening： study in both genders

Aim： To investigate whether urine is a good medium for screening and whether there is a correlation between the amount of extracted DNA and human papillomavirus （HPV）-positivity. Methods： In the present study, 30 first-voided urine （FVU） specimens and 20 urethroglandular swabs using cervex-brushes from male partners of HPV-positive patients, and 31 FVU specimens and 100 liquid-based cervix cytology leftovers sampled with cervix-brushes from HPV-positive women were examined for the presence of β-globin. Oncogenic HPV were detected using type-specific PCR. Results： β-globin was found in all the brushed samples, whereas it was found in only 68.9% of the FVU specimens. HPV-PCR was positive in 60.0% of the male brushes, in 29% of the female brushes and in 0% of the male FVU specimens. DNA concentration was, respectively, 0.9998 ng/μL, 37.0598 ng/μL and 0.0207 ng/μL. Conclusion： Urine is not a good tool for HPV detection, probably because the low DNA concentration reflects a low amount of collected cells. β-globin is measurable in FVU by real time quantitative PCR, but the DNA concentration is lower compared to brush sampling for both genders. β-globin-positivity of urethral and cervical swabs is 100%, showing a higher mean concentration of DNA, leading to a higher detection rate of HPV. This is the first article linking DNA- concentration to the presence of HPV.

DETECTION OF HUMAN PAPILLOMAVIRUS TYPES 16, 18 DNA RELATED SEQUENCES IN BRONCHOGENIC CARCINOMA BY POLYMERASE CHAIN REACTION

In studying the relationship between human papillomavirus (HPV) and bronchogenic carcinoma, 'high-risk' HPV 16, 18 DNA sequences were detected in samples from 50 lung cancer patients, 18 patients with benign pulmonary diseases and 4 fetal lung tissues by polymerase chain reaction (PCR) and dot-blot hybridization with biotin-labelled probes. The results showed that HPV 16, 18 DNA related sequences were found in 32% of lung cancer specimens, with 10 cases of HPV 16, 5 cases of HPV 18 and 1 case of both types. 48.15% (13 / 27) of squamous cell carcinomas were shown to be positive for HPV 16, 18 DNA. In addition, two adenocarcinomas and one small cell carcinoma were positive for HPV 16 DNA. No specimens from benign diseases tissues and fetal lung tissues showed positive results. These results suggest that primary bronchogenic carcinoma is related to HPV infection.

Asymptomatic Genital Infection of Human Papillomavirus in Pregnant Women and the Vertical Transmission Route

To further investigate the vertical transmission route of human papillomavirus (HPV) and the indication for the choice of mode of delivery, the infective status of 152 asymptomatic pregnant wemen and the maternal-fetal transmission were studied. By using general primers in polymerase chain reaction (GP-PCR) combined with restriction fragment length polymorphism analysis, HPV DNA positive rate in cervical secretions and venous blood in asymptomatic pregnant women was 36.21 % and 52.78 %, respectively, and the identified genotypes were mainly HPV_16 and _18. The maternal-fetal transmission rate of HPV via genital tract as well as blood was 40.91 % and 57.89 %, respectively. It was concluded that besides the transmission route of genital tract and amniotic fluid, there was also transplacental transmission of HPV in utero. Therefore,in our opinion, it is not an absolut indication to perform a cesarean delivery for the pregnant women with HPV asymtomatic genital infection.

THE DETECTION OF P53 GENE IN HUMAN CERVICAL CARCINOMA WITH AND WITHOUT HUMAN PAPILLOMAVIRUS INFECTION

The special primers Of p53 exon 7 as wed as HPV16 E6 and E7 ORFs (Opening Rending Frame) were used with PCR, PCR-SSCP technique, and 35 specimens of cervical carcinoma were examined. The results were as follows: ① HPV16 E6, E7 DNA was found in 25/35 specimens (71. 4%),which proved again HPV16 Infection an important event in cervical carcinogenesis. However only 11/35 (31.42% ) bad E6 and E7 ORFs simultaneously, 3/35 (8. 57%) and 11/35 (31. 42% ) had only E6 or E7 respectively. ② No mutation and LOH (Loss of Heterozygote) of p53 exon 7 were found in allof 35 specimens. Additionally in the present study, we developed a non-isotopic PCR-SSCP method.

Evaluation of Human Papillomavirus Infection among Women Using Pap Smear and PCR in Shiraz, Iran

Human papillomaviruses （HPVs） infect mucosal and cutaneous epithelia and cause malignancy and neoplastic lesions. These viruses cause 25,000 deaths per year from cervical cancer most often in developing countries. This major public health problem makes them important targets in the researches of papillomavirus detection methods. Since the early diagnosis of this virus infection would prevent neoplasia and cervical cancer, therefore in this study the combination of molecular and cytological methods were used to show the occurrence of the infection in women referred to Baghiatollah clinic of Shiraz. The results showed out of 110 cases, two samples were positive by PCR using GP5/6 primers but Pap smears showed only one sample of abnormal cytology. The rest 108 samples were negative by PCR and had normal cytology. The samples （1.82%） in evaluated women. present study showed a low occurrence of HPV infection in cervical

Head-to-head comparison of 7 high-sensitive human papillomavirus nucleic acid detection technologies with the SPF10 LiPA-25 system

Background:The SPF10 LiPA-25 system for human papillomavirus(HPV)detection with high analytical perfor-mance is widely used in HPV vaccine clinical trials.To develop and evaluate more valent HPV vaccines,other comparable methods with simpler operations are needed.Methods:The performance of the LiPA-25 against that of other 7 assays,including 4 systems based on reverse hybridization(Bohui-24,Yaneng-23,Tellgen-27,and Hybribio-16)and 3 real-time polymerase chain reaction(PCR)assays(Hybribio-23,Bioperfectus-21,and Sansure-26),was evaluated in selected 1726 cervical swab and 56 biopsy samples.A total of 15 HPV genotypes(HPV 6,11,16,18,31,33,35,39,45,51,52,56,58,59,and 66)were considered for comparison for each HPV type.Results:Among the swab samples,compared to LiPA-25,compatible genotypes were observed in 94.1%of samples for Hybribio-23,92.8%for Yaneng-23,92.6%for Bioperfectus-21,92.4%for Hybribio-16,91.3%for Sansure-26,89.7%for Bohui-24,and 88.0%for Tellgen-27.The highest overall agreement of the 15 HPV genotypes combined was noted for Hybribio-23(κ=0.879,McNemar’s test:P=0.136),followed closely by Hybribio-16(κ=0.877,P<0.001),Yaneng-23(κ=0.871,P<0.001),Bioperfectus-21(κ=0.848,P<0.001),Bohui-24(κ=0.847,P<0.001),Tellgen-27(κ=0.831,P<0.001),and Sansure-26(κ=0.826,P<0.001).Additionally,these systems were also highly consistent with LiPA-25 for biopsy specimens(all,κ>0.897).Conclusions:The levels of agreement for the detection of 15 HPV types between other 7 assays and LiPA-25 were all good,and Hybribio-23 was most comparable to LiPA-25.The testing operation of HPV genotyping should also be considered for vaccine and epidemiological studies.

细辛抗人乳头瘤病毒的作用研究

目的 :筛选细辛抗人乳头瘤病毒 (HPV)的有效部位。方法 :用系统溶媒 (石油醚、乙醚、乙酸乙酯、正丁醇、乙醇、蒸馏水 )对细辛依次进行提取 ,应用荧光定量聚合酶链反应技术 (FQ PCR)对各提取物进行体外抗病毒药效学试验 ,检测离体尖锐湿疣皮损中HPV DNA的扩增情况。结果 :HPV DNA经细辛水提物作用后 ,其PCR扩增检测为阴性 ,最低有效浓度为 0 4 g/ml,而细辛其它提取部位PCR扩增检测为阳性。结论

温州地区尖锐湿疣组织人乳头瘤病毒检测及型别分析

用人乳头瘤病毒（ＨＰＶ）通用引物（ＧＰ）及型特异性引物（ＳＰ）６、１１、１６、１８对４２例尖锐湿疣（ＣＡ）组织和４８例对照组进行多聚酶链反应（ＰＣＲ）检测。结果：ＣＡ组织五组引物扩增阳性率分别为８５．７％（３６／４２）、７３．８％（３１／４２）、１９．１％（８／４２）、２．４％（１／４２）、４．８％（２／４２），多重型别阳性率为１１．９％（５／４２）；对照组ＨＰＶ阳性率为６．３％（３／４８）。

复方杠柳液外用治疗尖锐湿疣临床观察及对离体人乳头瘤病毒DNA的影响

目的观察中药复方杠柳液治疗尖锐湿疣(condylomaacuminatum ,CA)的临床疗效,并从分子水平探讨其作用机理。方法将81例尖锐湿疣患者,随机分为复方杠柳液(30例)、杠柳浆(2 1例)、尤脱欣(30例) 3组,评定各组疗效及不良反应发生率。采用聚合酶链反应技术(polymerasechainreaction ,PCR) ,检测2例离体尖锐湿疣疣体中人乳头瘤病毒(humanpapillomavirus,HPV)DNA经复方杠柳液及其君药杠柳浆作用后的变化。结果复方杠柳液、杠柳浆、尤脱欣3组的痊愈率分别为56. 67%、42 . 86%、63 .33% ;总有效率分别为83 .33%、71. 43%、86. 67% ,3组疗效比较差异无显著性。复方杠柳液、杠柳浆、尤脱欣3组的不良反应发生率分别为6. 67%、4 .76%、86. 67% ,差异有显著性(P <0 . 0 1) ;复发率分别为14 .3%、12 . 5%、47. 1%。两份不同及相同浓度疣体匀浆混悬液经复方杠柳液、杠柳浆作用后PCR均阴性,未经药物处理的生理盐水组PCR均阳性。结论复方杠柳液、杠柳浆、尤脱欣3组最终疗效相近,复方杠柳液及杠柳浆组不良反应少、复发率低。该药对离体CA组织中的HPV- DNA可能有破坏作用。

疣毒净霜剂的研制及其体外药效学

目的 :研制治疗尖锐湿疣的疣毒净霜剂 ,并进行其抗人乳头瘤病毒 (HPV)的体外药效研究。方法 :以鸦胆子、紫草、莪术、细辛等为主药 ,制备水包油型霜剂。将霜剂作用于离体尖锐湿疣皮损组织中的HPV ,应用荧光定量聚合酶链反应 (FQ PCR)技术 ,通过定量检测HPV DNA的扩增情况 ,研究其抗病毒作用。结果 :经疣毒净霜剂作用后 ,DNA扩增受到抑制 ,检测结果为阴性 ;而对照组的DNA数则达每 1mL 1× 1 0 7拷贝 ,扩增没有受到抑制 ,结果为阳性。结论 :疣毒净霜剂制备工艺简单、稳定性好 ,具有明显的抗人乳头瘤病毒作用。

尖锐湿疣患者病损部位高危型人乳头瘤病毒的检测及其临床意义

目的了解尖锐湿疣(CA)患者高危型(16、18型)人乳头瘤病毒(HPV)的感染情况及其临床意义。方法应用实时荧光定量聚合酶链反应(FQ-PCR)对41例CA患者(扁平形16例,乳头形/菜花状25例)病损部位进行HPV16、18型DNA检测,又根据其发病次数分为初发和复发2组,分别有29例和12例,观察高危型HPV的感染率、病毒DNA含量及各组间的差异。结果CA患者高危型HPV的感染率为43.90%,与对照组(3.85%)比较差异有显著性(P<0.01)。扁平形和乳头形/菜花状CA患者高危型HPV的阳性率分别为18.75%和60.00%,2组比较也有明显差异(P<0.01);2组间病毒DNA含量比较差异无显著性(P>0.05)。初发和复发者HPV16、18型感染率及其DNA含量检测结果差异均无显著性(P>0.05)。25例乳头形/菜花状CA患者的疣体组织阳性率为36%,其表面分泌物的阳性率为52%。结论CA患者高危型HPV的感染率显著高于普通人群。乳头形/菜花状CA中高危型HPV感染率较扁平形明显升高。CA的复发与否可能与高危型HPV的感染无必然联系。同时检测疣体组织和分泌物的高危型HPV可提高检出率。

性病高危人群HPV DNA分型检测结果分析

目的分析性病高危人群HPV-DNA分型检测的结果。方法采用荧光定量聚合酶链反应技术(FQPCR)对541例性病高危人群进行HPV-DNA分型检测。结果男、女HPV阳性率分别为38.69%和52.63%,141例疣体中HPV阳性率为73.76%,其中男、女疣体HPV阳性率分别为61.54%和80.90%,400例分泌物中HPV阳性率为38.25%,其中男、女分泌物HPV阳性率分别为30.61%和42.69%,两者差异均具有显著性。配偶患CA者、CA治疗后复查者和自检者的HPV阳性率分别为44.12%,45.88%和33.33%。结论性病高危人群HPV检出率均较高,且女性高于男性。配偶患CA者、CA治疗后者和自检者也存在较高的HPV感染,应重视对这些人群的HPV常规检测。

CO_2激光碳化尖锐湿疣残存HPV DNA活性的实验研究

对20例尖锐湿疣(CA)患者,局部皮损经CO2激光治疗后,碳末DNA进行PCR扩增,并与正常角质形成细胞(KC)培养,观察KC增殖情况,观察其是否具有活性。结果发现:碳末DNAPCR扩增,40%阳性,碳末DNA对KC有明显抑制作用。从而推断碳末DNA仍有活性,可能是引起该病复发的原因之一。

柴胡对人乳头瘤病毒杀灭作用的实验研究

目的筛选柴胡抗人乳头瘤病毒(HPV)的有效成分。方法用溶媒(石油醚、乙醚、乙酸乙酯、正丁醇、乙醇、蒸馏水)对柴胡分别进行提取,应用荧光定量聚合酶链反应技术(FQ-PCR)对各提取物进行体外抗病毒药效学实验,检测离体尖锐湿疣皮损中HPV-DNA的扩增情况。结果HPV-DNA经柴胡水提物作用后,其PCR扩增检测为阴性,最低有效浓度为0.2g/mL,而用其他有机溶剂提取部分PCR扩增检测为阳性。结论柴胡的水提成分有抗人乳头瘤病毒作用。

应用PCR对尖锐湿疣和外耳道乳头状瘤中HPV DNA的分型检测

从手术切除的24例女性和12例男性尖锐湿疣新鲜标本中,以及42例女性尖锐湿疣、16例男性外耳道乳头状瘤和4例女性假性湿疣的石蜡包埋标本中,提取组织的基因组DNA,用人工合成的人乳头瘤病毒6.11和16型E6区特异性寡聚核苷酸引物,通过PCR进行HPV DNA的分型检测。结果66例女性尖锐湿疣中,感染HPV6型者4例,感染11型者12例,6+11型混合感染者49例;阴性1例,总检出率达98.4％。4例女性假性湿疣中1例为HPV6型感染,阳性率25％。16例男性外耳道乳头状瘤中HPV6+11型感染者5例,6+16型感染者3例,6+11+16型多重感染者8例,阳性率100％。12例男性尖锐湿疣中,HPV11型感染者7例,6+11型4例,阴性1例,总阳性率91.6％。还对细胞学上空泡化和非典型空泡化尖锐湿疣标本的HPV感染做了比较,未发现差异。

荧光定量聚合酶链反应在尖锐湿疣诊断中的应用

目的 探讨荧光定量聚合酶链反应 (FQ - PCR)对尖锐湿疣 (CA )的诊断价值。方法 FQ- PCR检测病理确诊的 CA患者 36份标本和健康人 84份标本 ;FQ- PCR及病理诊断同期检测临床送检病例 2 73份标本 ,并做诊断性试验评价。结果 36例病理确诊的 CA患者 HPV6和 HPV11的 DNA FQ- PCR全部阳性 ,平均拷贝数为 1.0× 10 7± 1.0× 10 2 / m l;84例健康人 FQ- PCR全部阴性 ,平均拷贝数为 3× 10± 2× 10 / ml。 FQ- PCR与病理诊断符合率为 10 0 % ,CA患者与健康人拷贝数的差异有显著性 (P<0 .0 5 )。FQ- PCR与病理诊断对比得出 :灵敏度为 10 0 % ,特异度为 92 % ,误诊率为 0 .0 8,漏诊率为 0 ,准确度为 98.9% ,阳性预测值为 98.8% ,阴性预测值为 10 0 % ,阳性似然比为 12 .5 ,阴性似然比为 0。结论 FQ - PCR能准确定量 ,灵敏度高 ,特异性强 ,快速 ,简便 。