原位聚合酶链反应技术检测结节性红斑皮损中Ⅱ型单纯疱疹...

目的 探讨结节性红斑（ＥＮ）的发病与Ⅱ型单纯疱疹病毒（ＨＳＶ－２）的关系。方法 用原位聚合酶链反应法（ＩＳＰＣＲ）,对３５ＮＥ皮损组织及１４例正常皮肤组织切片中的ＨＳＶ－２ＤＮＡ进行扩增,ＳＰ免疫组化染色,ＡＥＣ显色,结果 ＨＳＶ－２ＤＮＡ在ＥＮ及正常皮肤中的检出率分别为５７．１％（２０／３５）、８．３％（２／１４）,Ｐ〈０．０１,阳性信号主要位于皮损部位,以小血管为著,细胞内的分布以核内型为主,     

聚合酶链反应检测多形红斑皮损中单纯疱疹病毒DNA

为了研究多形红斑（ＥＭ）与单纯疱疹病毒（ＨＳＶ）之间的关系，特别是探讨与疱疹相关的ＥＭ的临床特点及其在ＥＭ发病中所占比例，应用聚合酶链反应（ＰＣＲ）方法检测了６０例ＥＭ的典型皮损的活检组织和３２例正常组织及疾病对照标本中的ＨＳＶＤＮＡ。结果显示：２１例（３５％）ＥＭ患者检测出ＨＳＶ特异ＤＮＡ，对照标本（１６份正常皮肤组织和１６份其它皮肤病的皮损标本）中未发现ＨＳＶＤＮＡ。根据病史提供，６０例ＥＭ中有５例是药物相关的ＥＭ，８例是病毒相关的ＥＭ，其余４７例为特发型ＥＭ。研究表明：２１例ＰＣＲＨＳＶＤＮＡ阳性的ＥＭ患者中，仅有２例发病前有口唇疱疹病史，１５／２１例（７１．４％）是复发性ＥＭ，发病高峰在冬季。尽管临床上无明显ＨＳＶ感染史，多数特发性ＥＭ也与疱疹病毒感染有关。ＰＣＲ技术是检测ＥＭ皮损中ＨＳＶ的快速、有效的方法，它可以决定病因上和ＨＳＶ感染有关的病例。     

用聚合酶链反应检测多形红斑皮损中单纯疱疹病毒的DNA

用聚合酶链反应检测２３例多形红斑石蜡组织切块中ＨＳＶ－ＤＮＡ，１６例为阳性（６９．５６％），其中ＨＳＶ－Ⅰ型１３例（８１．２５％），ＨＳＶ－Ⅱ型３例（１８．７５％）。对照组为银屑病３例，大疱性类天疱疮５例，扁平苔藓３例均为阴性。对于探讨ＨＳＶ在多形红斑发病机理中的作用有重要意义。     

应用原位聚合酶链反应检测淋病患者HSV-Ⅱ感染情况的报告

为了解淋病患者ＨＳＶ Ⅱ的感染情况，应用原位聚合酶链反应（ＩＳＰＣＲ）和ＰＣＲ技术对５８例淋病患者进行ＨＳＶ Ⅱ检测。结果两种方法ＨＳＶ ⅡＤＮＡ阳性检出率均为５３．４５％（３１／５８）。ＩＳＰＣＲ显示，１１例有皮损者中，９例阳性信号存在于上皮细胞核及胞浆，既往有疱疹皮损者６例，阳性信号均存在于细胞核，４１例无明显皮损者，１６例阳性，其中１２例阳性信号在细胞核，４例在胞核及胞浆。结果提示，淋病患者合并有高的ＨＳＶ Ⅱ感染率。应用ＩＳＰＣＲ不仅可以敏感特异地检测ＨＳＶ Ⅱ，而且可以定位，为临床诊断和治疗提供重要依据     

STD门诊患者血清HSV－2 IgG抗体检测报告

采用ＥＬＩＳＡ技术检测了８０例ＳＴＤ门诊患者（其中淋病１４例、非特异性泌尿生殖道感染３８例及尖锐湿疣２８例）血清单纯疱疹病毒２型（ＨＳＶ－２）ＩｇＧ抗体。结果发现ＳＴＤ患者血清ＨＳＶ－２ＩｇＧ抗体阳性率为６０．０％，远比正常人群中的２５．３％为高（Ｐ＜０．００１），说明生殖器疱疹病毒感染在我国与西方国家一样，有上升和流行趋势。     

STD门诊患者血清HSV—2IgG抗体检测报告

采用ＥＬＩＳＡ技术检测了８０例ＳＴＤ门诊患者（其中淋病１４例、非特异性泌尿生殖道感染３８例及尖锐湿疣２８例）血清单纯疱疹病毒２型）ＨＳＶ－２）ＩｇＧ抗体。结果发现ＳＴＤ患者血清ＨＳＶ－２ＩｇＧ抗体阳性率为６０．０％，远比正常人群中的２５．３％为高（Ｐ＜０．００１），说明生死器疱疹病毒感染在我国与西方国家一样，有上和流行趋势。     

性病门诊患者人类免疫缺陷病毒和单纯疱疹病毒感染的血清学检测

为了解我同性病患者中人类免疫缺陷病毒（ＨＩＶ）和单纯疱疾病毒（ＨＳＶ）感染的流行情况，我们选择了我所性病门诊的病人进行了淋球菌分离培养以及ＨＩＶ和ＨＳＶ感染的血清学检测。结果表明，８８份临床标本淋球菌分离培养阳性率为２８．４％。ＨＳＶ－１和ＨＳＶ－２抗体检出率分别为５９．１％和２５．０％。统计分析结果表明，淋球菌感染和ＨＳＶ－２抗体阳性之间有一定的相关性（Ｐ＜０．０１）。８８份血清标本ＨＩＶ抗体检出率为０．     

性病门诊患者人类免疫缺陷病毒和单纯疱疹病毒感染的血清学检测

为了解我同性病患者中人类免疫缺陷病毒（ＨＩＶ）和单纯疱疾病毒（ＨＳＶ）感染的流行情况，我们选择了我所性病门诊的病人进行了淋球菌分离培养以及ＨＩＶ和ＨＳＶ感染的血清学检测。结果表明，８８份临床标本淋球菌分离培养阳性率为２８．４％。ＨＳＶ－１和ＨＳＶ－２抗体检出率分别为５９．１％和２５．０％。统计分析结果表明，淋球菌感染和ＨＳＶ－２抗体阳性之间有一定的相关性（Ｐ＜０．０１）。８８份血清标本ＨＩＶ抗体检出率为０．     

生殖器疱疹的诊断及感染特征探讨

细胞培养法对８０例临床诊断为典型生殖器疱疹的患者进行了人类单纯疱疹病毒（ＨＳＶ）分离，并结合链霉亲和素过氧化物酶免疫组化法（ＬＳＡＢ）及ＰＣＲ法对其中部分患者进行了ＨＳＶ及其型别检查。三种方法的ＨＳＶ检出率分别为５８．８％、２０％、９０％；培养法作为金标准特异性高，对早期患者检出率达８６．４％；ＰＣＲ法与其他两法比较敏感性高，并利于ＨＳＶ型别的诊断；ＬＳＡＢ法对临床标本的检出率虽很低，却可提高培养法的检出率。ＨＳＶ的感染型别ＨＳＶ２占９１．７％、ＨＳＶ１占８．３％。还讨论了ＰＣＲ引物的优点，并总结了本组患者的临床感染特征。     

原位聚合酶链反应技术检测结节性红斑皮损中Ⅱ型单纯疱疹病毒DNA

目的　探讨结节性红斑（ Ｅ Ｎ） 的发病与Ⅱ型单纯疱疹病毒（ Ｈ Ｓ Ｖ２） 的关系。方法　用原位聚合酶链反应法（ Ｉ Ｓ Ｐ Ｃ Ｒ） ,对３５ 例 Ｅ Ｎ 皮损组织及１４ 例正常皮肤组织切片中的 Ｈ Ｓ Ｖ２ Ｄ Ｎ Ａ 进行扩增, Ｓ Ｐ 免疫组化染色, Ａ Ｅ Ｃ 显色。结果　 Ｈ Ｓ Ｖ２ Ｄ Ｎ Ａ 在 Ｅ Ｎ 及正常皮肤中的检出率分别为５７ ．１ ％ （２０／３５） 、８ ．３ ％ （２／１４） , Ｐ＜ ０ ．０１ ,阳性信号主要位于皮损部位,以小血管为著,细胞内的分布以核内型为主,亦有核浆混合型。结论　 Ｅ Ｎ 的发病与 Ｈ Ｓ Ｖ２ 有关。     

性病门诊836例患者单纯疱疹Ⅰ,Ⅱ型检测结果分析

目的探讨性病专科门诊患者中HSV-Ⅰ,Ⅱ型病毒的感染情况及在典型和非典型生殖器疱疹(GH)患者中的分布。方法采用荧光定量聚合酶链反应技术(FQ-PCR)对836例性病专科门诊患者进行HSV-Ⅰ,Ⅱ型DNA的检测。结果836例患者中,HSV-Ⅰ型感染38例(4.54%),男性阳性率5.20%,女性为3.36%,二者差异无显著性;HSV-Ⅱ型感染380例(45.45%),男性阳性率39.96%,女性为55.37%,二者差异有显著性(P<0.05)。具有典型GH或高度可疑GH症状者369例,HSV感染者218例;无明显GH症状者467例,HSV感染者198例,二者差异有显著性(P<0.05)。结论HSV在本地区具有较高感染率,以HSV-Ⅱ型为主。在非典型GH患者中,HSV的检出率较高,须引起临床注意。     

女性原发性生殖器疱疹中HSV-Ⅱ的检测及意义

目的探讨单纯疱疹病毒Ⅱ型(HSV-Ⅱ)与女性原发性生殖器疱疹(GH)的相关性及意义。方法应用酶联免疫吸附法(ELISA)分别检测患者血清中HSV-Ⅱ抗体以及分泌物中的HSV-Ⅱ抗原。结果HSV-Ⅱ中IgG均阴性的138例女性生殖器疱疹患者中HSV-Ⅱ抗原阳性78例(56.52%);HSV-Ⅱ中IgM抗体阳性130例(94.20%),抗体阳性率明显高于抗原阳性率(P<0.01)。典型病例组抗原阳性65例(87.84%),IgM抗体阳性66例(89.19%);不典型病例组抗原阳性13例(20.31%),IgM抗体阳性64例(100.00%)。结论对于皮损时间短,症状典型者可检测HSV-Ⅱ抗原;皮损时间长,或反复者可检测HSV-Ⅱ抗体,可以有效提高HSV感染的临床诊断率。     

寻常型银屑病与单纯疱疹病毒1型相关性的研究

目的 探讨寻常型银屑病与单纯疱疹病毒1型(HSV-1)的可能相关性。方法 应用PCR法检测患者皮损、外周血单一核细胞(PBMCs)和咽拭子中HSV-1DNA,ELISA法检测患者血清中抗HSV-1的IgM、IgG抗体,并与正常人对照做比较。结果 患者皮损、PBMCs和咽拭子中HSV-1DNA检出率分别为37.5%、18.6%和18.8%,血清中抗HSV-1的IgM、IgG抗体检出率分别为37.2%和53.5%.经χ²检验,患者皮损、PBMCs中HSV-1DNA和血清中IgM抗体检出率显着高于正常人对照,点滴状患者的皮损、PBMCs和咽拭子中HSV-1DNA以及血清中抗HSV-1IgM抗体检出率显着高于斑块状患者。结论 寻常型银屑病尤其是皮损呈点滴状者与HSV-1显着相关,患者可能存在HSV-1的近期感染。     

彩色乳胶免疫层析法检测单纯疱疹病毒1,2型抗原临床应用分析

目的探讨彩色乳胶免疫层析法检测单纯疱疹病毒(HSV)1和2型抗原的临床应用价值。方法采用彩色乳胶颗粒免疫层析法检测各种水疱、溃疡和其他皮损中的HSV-1和2型抗原,同时用实时荧光定量PCR法进行对照。结果免疫层析法可检出≥1×106DNA拷贝/mL的HSV-1型混悬液和≥5×105DNA拷贝/mL的HSV-2型混悬液。132例标本中,免疫层析法检出HSV阳性43例,PCR检出HSV阳性50例。以PCR法为金标准,免疫层析法的灵敏度、特异度、阳性预测值和阴性预测值分别为84.00%,98.78%,97.67%和91.01%。结论免疫层析法检测HSV敏感性和特异性高,具有方便、快速和经济的特点,适于有症状患者的及时检测,对疱疹感染早期诊断和及时治疗有重要作用。     

Seroprevalence of HSV-1 and HSV-2 infection in the general French population

BACKGROUND: In spite of the large prevalence and growing incidence of herpes simplex infection (HSV-1 and HSV-2), relatively few large serological surveys are available worldwide and it is still difficult compare frequencies of HSV contaminations in various countries. We present the results of HERPIMAX, the first epidemiological inquiry on HSV prevalence in the general French population. METHODS: Of a cohort of 12,735 presumed healthy adult volunteers included in the prospective study SU.VI.MAX, designed to assess the relation between nutritional supplementations and degenerative diseases, HERPIMAX randomly selected 4412 subjects (females 66.5%, males 33.5%). All serum samples were assessed for HSV-1 and HSV-2 IgG antibodies with a HSV type specific, enzyme immunosorbent assay (EIA). Equivocal result were retested with another HSV type specific immunoblot assay combined with a type common HSV IgG EIA in order to give a definitive interpretation. RESULTS: The mean seroprevalence was 67% for HSV-1 and 17.2% for HSV-2. For HSV-2 the seroprevalence was higher in females (17.9%) compared with males (13.7%) (p<0.001). For both HSV types, there was no significant difference in prevalence as regards age distribution in males and females, whereas prevalence increased significantly with age in females for HSV-1. Univariate analysis showed a significant association between HSV-1 prevalence and education level in males and females (p<0.001) and between HSV-2 prevalence and marital status in both sexes (p<0.001). There were geographical disparities, with a higher HSV-2 prevalence in the south of France as well as in Paris. CONCLUSION: These results confirm a high prevalence of HSV infection in France. They are also in agreement with previous results of other survey carried out in other developed countries as regards higher prevalence of HSV-2 infection in women, the stability of seroprevalence for both HSV types after 35 years of age in females and 45 years of age in males.     

Viral DNA detection and RAS mutations in actinic keratosis and nonmelanoma skin cancers

Background Actinic keratosis (AK) is a well‐established precancerous skin lesion that has the potential to progress to squamous cell carcinoma (SCC). Basal cell carcinoma (BCC) is a locally aggressive slowly growing tumour that rarely metastasizes. A number of viruses have been proposed to play a role in the development of nonmelanoma skin cancers (NMSC), but the most plausible evidence to date suggests that cutaneous human papillomavirus (HPV) is the key instigating factor. Objectives To evaluate the prevalence of HPV, cytomegalovirus (CMV), herpes simplex virus (HSV) and Epstein–Barr virus (EBV) and investigate their relationship with the presence of RAS gene mutations in cutaneous lesions obtained from nonimmunosuppressed patients. Methods HPV, CMV, HSV and EBV detection was performed using polymerase chain reaction (PCR) in skin biopsies (26 AK, 12 SCC and 15 BCC samples) that were collected from immunocompetent patients. The RAS mutation incidence was also investigated in all cutaneous lesions by use of PCR/restriction fragment length polymorphism and direct DNA sequencing. Results Seventeen out of 53 (32%) skin lesions were found to be positive for HPV DNA. The highest incidences of HPV infection were five of 15 (33%) in BCC and four of 12 (33%) in SCC specimens. The HPV incidence was eight of 26 (31%) in AK and eight of 53 (15%) in normal skin tissue. Twelve out of 53 (23%) skin lesions were CMV‐positive. The highest incidence of CMV infection was six of 15 (40%), observed in BCC specimens. The CMV incidence was two of 26 (8%) in AK and four of 12 (33%) in SCC. No normal skin biopsy was found to be positive for CMV. All cutaneous samples were negative for HSV and EBV DNA, as assessed by our PCR‐based assays. Only three samples, one AK (4%), one BCC (6%) and one SCC (8%), were found to carry a G>T transversion at the second position of HRAS codon 12. Both HRAS mutant SCC and BCC biopsies were HPV‐ and CMV‐positive, as well. Conclusions HPV DNA is detected in NMSC, AK and normal skin biopsies. Our results also indicate that CMV is involved in NMSC at higher levels than in premalignant lesions, whereas the virus was not detected in normal skin biopsies. HSV and EBV do not appear to be involved in the pathogenesis of cutaneous lesions. Moreover, we suggest that the HRAS codon 12 mutation is not a very common event in AK or NMSC. Finally, both viral infection and HRAS activation appear to represent independent factors in the aetiology of NMSC, samples of which were obtained from immunocompetent patients.     

生殖器部位皮损的单纯疱疹病毒检测及分型

目的 探讨生殖器疱疹部位皮损的不典型表现及其与单纯疱疹病毒型别的关系。方法 对外生殖器部位及其周围有硬结或疖肿、裂隙、毛囊炎等非水疱性皮肤黏膜损害的患者进行临床资料采集和分析,并对皮损标本进行单纯疱疹病毒的分离培养、PCR检测和病毒分型。结果 105例有外生殖器部位非水疱性皮损的患者入选本研究,在硬结(或疖肿)、裂隙、毛囊炎、类似擦破、单个溃疡、非特异性红斑和红肿渗液性包皮龟头炎皮损中,PCR检测HSV的阳性率分别33.3%(6/18)、20%(3/15)、37.5%(6/16)、28.6%(2/7)、33.3%(4/12)、20%(5/25)和50%(6/12),总的检出阳性率为30.5%(32/105)。分离培养法检测HSV的阳性率分别为22.2%(4/18)、13.3%(2/15)、25%(4/16)、14.3%(1/7)、33.3%(4/12)、8%(2/25)和41.7%(5/12),总的检出阳性率为21%(22/105)。两种方法检测HSV的总检出率差异无统计学意义(κ=0.095,P=0.114)。HSV-PCR分型结果与荧光单克隆抗体分型结果相符。在所有HSV阳性者中,HSV-1感染占9.4%(3/32),HSV-2感染占90.6%(29/32)。结论 生殖器HSV感染的皮肤黏膜损害多样,可为外生殖器部位的硬结(疖肿)、裂隙、毛囊炎、类似擦破、单个溃疡、非特异性红斑和红肿渗液性包皮龟头炎等不典型表现,而且主要由HSV-2感染引起。     

初发性生殖器疱疹患者的临床特征及复发因素分析

目的 分析单纯疱疹病毒2型(HSV-2)所致的初发性生殖器疱疹(GH)患者的临床特征及复发的影响因素.方法 选取2015年1月至2019年12月广州医科大学附属市八医院皮肤性病科门诊确诊的189例初发性GH患者作为研究对象,对其临床资料、实验室检查结果进行回顾性分析,对临床特征和可能影响其复发的因素进行相关性分析.结果...     

Detection of herpes simplex virus genomic DNA in various subsets of Erythema multiforme by polymerase chain reaction

BACKGROUND: The wide variation in the detection of herpes simplex virus (HSV) DNA (36-75%) by polymerase chain reaction (PCR) in erythema multiforme (EM) may be partly attributed to differences in case selection in terms of subsets of EM studied. OBJECTIVE: To determine the frequencies of detection of HSV DNA in specific subsets of EM. METHODS: Nested PCR was used to detect HSV DNA in skin biopsies with histologically proven EM. RESULTS: PCR was performed on skin biopsies from 63 patients with EM. HSV DNA was detected in 3/11 (27.2%) of single-episode HSV-associated EM (HAEM), 6/10 (60%) of recurrent HAEM, 1/4 (25%) of single-episode idiopathic EM and 6/12 (50%) of recurrent idiopathic EM. HSV DNA was not detected in atypical EM (0/11), suspected drug-induced EM (0/9) or Stevens-Johnson syndrome (0/6). CONCLUSION: The overall PCR positive rates of HAEM (42.9%) and idiopathic EM (43.8%) were comparable suggesting that idiopathic EM is likely to be related to a subclinical HSV infection.     

定量PCR和间接免疫荧光法联合检测生殖器疱疹病毒感染

目的：研究间接免疫荧光法（IIF）检测生殖器HSV感染的敏感性和特异性。方法：以定量PCR为对照,用HSV型共同性糖蛋白单克隆抗体为夹心的IIF法,检测了94例临床诊断为生殖器疱疹的患者皮疹中的HSV。结果：IIF法检测HSV的敏感性为74．12％,特异笥为55．60％;总阳性率（71．30％）,明显低于定量PCR法的阳性率（90．40％）（P＜0．05）,但两种方法检测GH患者皮水疱内的HSV阳性率无明显差异（86．20％vers.97.00%),而检测糜烂性皮疹内的HSV时,PCR法的阳性率高于IIF法（P＜0．05）,结论：IIF法具有简单,快捷的优点,适用于检测早期可疑GH患者皮疹内HSV,有临床实用价值。