自贡地区女性15种高危型人乳头瘤病毒感染情况分析

目的了解自贡地区15种高危型人乳头瘤病毒(HR-HPV)基因型感染的分布情况,为自贡地区HR-HPV分子流行病学研究提供科学依据。方法对371例宫颈分泌物标本采用多通道实时荧光定量PCR技术检测15种HR-HPV基因亚型,分析不同年龄段女性HR-HPV感染情况。结果 371例临床标本中共检出阳性56例,检出率15.09%;单一HR-HPV基因亚型感染41例(11.05%),复合感染15例(4.04%);其中HPV16、52、58、18是最常见的高危型,检出率分别为5.93%、3.23%、2.96%、2.43%。HR-HPV感染自贡女性人群的年龄分布以18~50岁年龄段为主,其中18~30岁年龄段的女性人群检出率最高(19.30%),高于其余年龄段,差异无统计学意义(P0.05)。结论自贡地区15种HR-HPV感染有年轻化趋势,HR-HPV基因型检测对于自贡地区HR-HPV感染的诊疗、预防性疫苗的使用及宫颈癌的早期干预和预防具有重要意义。     

多重实时PCR检测高危型人乳头瘤病毒感染

目的 调查不同宫颈病变组妇女生殖道13种高危型HPV(HR HPV)感染情况,分析HR HPV感染与宫颈病变的关系.方法 选择深圳市宝安区妇幼保健院宫颈专科门诊患者350例,利用TCT技术检测其宫颈上皮细胞学状况,根据细胞学结果分组.采用多重实时PCR(mRT PCR)检测其生殖道HR HPV感染情况和病毒载量,采用χ2检验或Fisher精确概率法比较各组间HR HPV感染率,采用Kruskal-Wallis或Wilcoxon秩和检验比较各组间HR HPV阳性者的病毒载量,采用Wilcoxon秩和检验比较HR HPV阳性和阴性组的年龄分布情况.结果 未见上皮内病变或恶性病变(NILM)组、意义不明的非典型鳞状上皮细胞增生(ASCUS)组、鳞状上皮细胞低度病变(LSIL)组、鳞状上皮细胞高度病变(HSIL)组的HR HPV阳性例数分别为10例(3.4%)、7例(20.0%)、11例(78.6%)和6例(6/6).NILM组HR HPV阳性率低于ASCUS组、LSIL组,ASCUS组HR HPV阳性率低于LSIL组,差异有统计学意义(χ2值分别为14.43、107.69、14.76,P均<0.01);NILM组低于HSIL组、ASCUS组低于HSIL组,差异有统计学意义(P值分别为0.000 1、0.000 4).NILM组、ASCUS组、LSIL组和HSIL组HR HPV阳性者的病毒载量分别为4.10 (3.38～6.27)、5.33 (3.63～6.66)、5.77 (4.01～7.01)和5.58 (4.19～5.85)(拷贝/ml,lg).将ASCUS组、LSIL组和HSIL组3组合并为宫颈细胞异常组,病毒载量为5.58(3.63～7.01)(拷贝/ml,lg)高于NILM组,差异有统计学意义(U=43.0,P<0.05).HR HPV阳性组平均年龄36(21～56)岁,与阴性组[33(21～58)岁]比较,差异无统计学意义(U=4 544,P>0.05).结论 HR HPV感染与宫颈病变有关;HR HPV病毒载量高低与宫颈病变程度无相关性,但病毒载量越高,存在宫颈病变的可能性越大.HR HPV阳性组与阴性组的年龄分布无差异.

Abstract：

Objective To investigate 13 high-risk types of HPV (HR HPV) infection rates in women with different grades of cervical lesions.Methods A total of 350 women, who were hospitalized in the department of gynecology in Bao′an Maternity & Child health hospital, were enrolled for the study.TCT technology was used to evaluate the cervical epithelium.The group were divided according to the cytology results.Multiplex real time PCR (mRT PCR) was used to detect the viral loads.HR HPV infection rate of different groups were analyzed using χ2 test or Fisher exact test.HR HPV viral loads of patients in different grades of cervical lesion groups were compared using Kruskal-Wallis or Wilcoxon test, and the age distribution of HR HPV positive group and negative group was analyzed by using Wilcoxon test.Results The HR HPV infection rates of NILM, ASCUS, LSIL, HSIL were 3.4% (10/295), 20.0% (7/35), 78.6% (11/14) and 100.0% (6/6), respectively.HR HPV positivity in NILM was lower than ASCUS (χ2=14.43,P<0.01) and LSIL (χ2=107.69,P<0.01), HR HPV positivity in ASCUS was lower than LSIL (χ2=14.76,P<0.01). The median of HR HPV viral loads in NILM, ASCUS, LSIL and HSIL were 4.10 (3.38-6.27), 5.33 (3.63-6.66), 5.77 (4.01-7.01) and 5.58 (4.19-5.85) respectively (copies/ml,lg).Combined ASCUS, LSIL and HSIL groups into cervical lesion group, HR HPV viral load of which was higher than that of NILM (U=43.0, P<0.05).The median Ages of HR HPV positive group and negative group were 36 and 33, respectively.No statistical significance was found between them (U=4 544, P>0.05).Conclusions The present study revealed that HR HPV infection was related to cervical lesion, but there was no correlation between viral load and cervical lesion grade. In additional, no difference in age distribution was found between HR HPV positive group and negative group.     

第二代杂交捕获法与多重荧光PCR在检测高危型人乳头瘤病毒中的应用比较

目的探讨第2代杂交捕获法（HC2）和多重荧光聚合酶链反应（PCR）在女性生殖道高危型人乳头瘤病毒（HPV）中的检测性能。方法随机选择267例宫颈疾病患者，采集宫颈部位分泌物，运用美国食品药品监督管理局（FDA）认证的HC2和多重荧光PCR分别检测标本中的HPV，并将2种方法检测结果不一致的标本进行测序分析。结果HC2和多重荧光PCR检测高危型HPV的阳性率分别为33．33％和31．46％；HC2和多重荧光PCR对高危型HPV感染的诊断敏感性分别为85．88％和90．32％，特异性分别为91．21％和99．43％。结论与HC2相比，多重荧光PCR检测高危型HPV的敏感性和特异性均有提高。     

妇女高危型人乳头瘤病毒核酸定量检测结果分析

目的探讨阳江地区妇女高危型人乳头瘤病毒（HPV）感染情况和年龄分布。方法选取2012年5月至2013年4月在该院妇科就诊的1744例妇女宫颈棉拭子标本,采用实时荧光定量PCR技术检测高危型HPVDNA。将筛查人群分成10个年龄组,分别计算各年龄组高危型HPV阳性率。结果1744例妇女中,共检测出高危型HPV阳性者200例,总阳性率为11．5％（200／1744）,高危型HPV阳性者在大于40~45岁和大于35-40岁2个年龄组的分布率较高,分别是25．O％（50／200）和22．5％（45／200）。结论对妇女进行HPV检查,可为宫颈癌的早期诊断、预防及治疗效果提供实验室诊断依据。     

喉乳头瘤中人乳头瘤病毒DNA的PCR检测

喉乳头瘤中人乳头瘤病毒ＤＮＡ的ＰＣＲ检测董菁，庄坚，余秀葵（汕头大学医学院微生物学教研室，广东汕头５１５０３１）关键词乳头瘤，人乳头瘤病毒，聚合酶链反应喉乳头瘤（Ｌａｒｙｎｇｅａｌｐａｐｉｌｌｏｍａ，ＬＰ）为耳鼻喉科常见病之一，该病虽可自发缓解，但易...     

高危人乳头瘤病毒检测在实时多重荧光核酸扩增技术中的意义

目的 探讨实时多重荧光聚合酶链反应(PCR)检测宫颈脱落细胞内高危型人乳头瘤病毒(HPV)感染的临床意义,建立快速、准确、适于推广的检测方法.方法 采用实时多重荧光PCR技术检测1 560例宫颈脱落细胞高危型HPV(16、18、31、33、35、39、45、51、52、56、58、59、68)感染状况.结果 1 560例患者高危型HPV阳性率为21%(327/1 560).其中宫颈炎、湿疣、LSIL 、HSIL 、鳞状细胞癌患者的高危型HPV阳性率分别为10.3%、28.4%、21.6%、35.2%和88.9%.结论 实时多重荧光PCR技术检测宫颈脱落细胞内高危型HPV感染,具有快速、简便、灵敏、不易污染等优点,适用于大规模筛查和高危型HPV持续感染监测.     

人乳头瘤病毒PCR产物酶标-微孔板杂交分型

目的建立聚合酶链反应（ＰＣＲ）产物直接酶标记微孔板反向分子杂交法用于人乳头瘤病毒（ＨＰＶ）型别鉴定。方法利用ＨＰＶ通用引物介导ＰＣＲ（ＧＰＰＣＲ）扩增靶ＤＮＡ，然后对产物直接标记辣根过氧化物酶复合物（ＨＲＰＰＥＩＱＩ），与预先包被在微孔板上的ＨＰＶ寡核苷酸探针杂交后酶显色法检测。结果ＨＰＶ各型之间无交叉杂交；杂交灵敏度可达１３～７６个病毒ＤＮＡ拷贝，比ＰＣＲ琼脂糖凝胶电泳检测高１０倍；该法重复性好，阳性孔批内ＣＶ为６．３４％，批间ＣＶ为１０．５３％，阴性孔批内ＣＶ为１％，批间ＣＶ为５．７９％；而且杂交影响因素较少。结论该法特异性强、灵敏度高、操作简便，无放射性和ＥＢ染料污染，杂交时间短、仪器读取结果，便于大量常规临床样本定性或定量检测。     

不同引物介导的聚合酶链反应对人乳头瘤病毒的快速检测与分型

目的 比较通用引物、型特异性引物在检测人乳头瘤病毒（HPV）DNA中的应用。方法 104例尖锐湿疣组织中提取DNA后分别用HPV6b、11、16、18型特异性引物及通用引物MY09／11、GP5^＋／6^＋经聚合酶链反应（PCR）进行扩增。部分标本采用限制性内切酶Pst I来确定HPV11型。结果 GP5^＋／6^＋的检出率高于MY09／11，但差异无显著性（χ^2=3．78，P〉0．05）。GP5^＋／6^＋检出微量HPV DNA的敏感度高于MY09／11。型特异性引物检测HPV感染优于通用引物MY09／11结合酶切分型。结论 型特异性引物检出HPV11单一型感染显著多于HPV6b型，且发现HPV6b、11型同时感染有逐年增高之趋势。GP5^＋／6^＋引物结合型特异性引物可为临床上检测尖锐湿疣HPV型别提供方便易行的选择。     

1294例女性高危型人乳头瘤病毒基因分型结果回顾性分析

目的了解北京地区妇科门诊就诊女性高危型人乳头瘤病毒(HPV)的感染及其基因亚型分布情况,为该市今后防治人乳头瘤病毒感染和宫颈癌提供参考依据。方法回顾性分析2013年1月至2014年5月该院妇科门诊就诊的1 294例女性宫颈拭子的13种高危型HPV基因分型检测结果,比较不同基因型的流行病学特点。采用SPSS17.0对数据进行统计学分析。结果 1 294例妇科门诊就诊女性中,以58型、16型和52型HPV最为常见,检出率分别为10.5%、9.2%和8.2%。各年龄段就诊女性中,30~40岁的HPV感染率最高(39.9%),其次为40~50岁、大于或等于60岁,差异无统计学意义(P0.05)。结论该地区妇科门诊就诊女性高危型HPV感染率较高,应加强HPV筛查力度,为今后HPV相关疾病的防治提供基础依据。     

荧光定量PCR检测在宫颈癌筛查中的应用

目的探讨荧光定量聚合酶链反应(FQ-PCR)检测宫颈病变人乳头瘤病毒(HPV)13种高危型DNA的可行性,为检测宫颈组织中HPV的感染提供有效的方法。方法选择197例阴道细胞学检测异常的宫颈脱落细胞,使用FQ-PCR方法检测13种高危型HPV DNA,同时取宫颈病变处组织进行组织病理学检查。结果 329例样本中HPV 13种高危型阳性者218例,阳性率为66.26%:CT值范围在13～40之间,平均为(26±2.31);组织病理学检测186例阳性,阳性率为56.53%,经χ~2检验,差异有统计学意义(χ~2=6.566,P=0.0104);阴性标本为Undet。结论 FQ-PCR方法检测宫颈病变中13种高危型HPV DNA感染,具有快速、简便、灵敏度高、特异性强等优点,对宫颈癌的普查和治疗有指导意义。     

基因芯片检测技术在女性生殖道HPV分型感染中的应用

目的探讨女性生殖道人乳头瘤病毒（HPV）各种基因分型感染的情况。方法采用HPV分型基因芯片检测系统,同时检测5种低危型HPV亚型和18种高危型HPV亚型。结果测定250例标本中HPV阳性92例（36．8％）,20～35岁的年龄阶段检出率最高（占总阳性数73．9％）,其中感染单一亚型57例（61．9％）,感染2种亚型28例（30．4％）,感染3种亚型及以上7例。低危型主要为HPV6、11,其次为43;高危型主要为16、18、33、35、45、31、58等亚型。结论基因芯片对于检测HPV的多种型别和HPV多型别混合感染具有重要意义,可为HPV感染和宫颈癌的防治提供重要依据。     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.     

杂交捕获法和PCR-RDB法检测妇女HPV感染的比较

Objective To compare hybrid capture-Ⅱ(HC2)to polymerase chain reaction(PCR)-reverse dot-blot(RDB)for detecting HPV infection in women.Methods HC2 method and PCR-RDB were applied to detecting human papillomaviral infection in female genital duct,and the test results were analyzed.Results Concordant results were found in 81.14%(400/493)samples(Kappa=0.63;95%CI,0.57-0.69).The high risk type HPV positive rate detected by PCR-RDB were 1.60%(3/187),29.85%(20/67),69.88%(58/83),86.52%(77/89)and 91.04%(61/67),respectively,when the HC2 RLU/CO of samples were＜1.00,1.00-9.99,10.00-99.99,100.00-999.99 and≥1000.00.Statistical differences were found among the five groups(χ2=289.3,P＜0.01).Cross reactions were found between HC2 high risk probe and HPV53,66,11,cp8304 and other genotypes.Conclusion There is good concordance between HC2 and PCR-RDB,though cross reaction exists in HC2.Relatively speaking,there are more influential factors for PCR-RDB.