多聚酶链反应与酶标法检测生殖器疱疹病毒感染的比较研究

为探讨多聚酶链反应(PCR)与酶标法检测生殖器疱疹病毒感染的诊断价值。采用PCR和酶标法检测生殖器疱疹病毒感染者45例，并用PCR作病毒分型。结果：临床确诊的45例患者中，酶标法检测抗原HSV阳性27例，阳性率60．0%(27／45)；PCR检测HSV阳性42例，阳性率93．33％(42／45)．其中HSV-11例，占2．38%；HSV-241例，占97．62％。对于有皮损的患者，PCR法较酶标法有较高的阳性检出率。由此可见：(1)HSV-2是主要的致病因素；(2)在对病毒分型及有皮损患者的诊断上，PCR法明显优于酶标法。     

酶联免疫吸附试验在检测生殖器疱疹病毒抗原中的应用

目的探讨酶联免疫吸附试验(ELISA)检测泌尿生殖道分泌物及皮损中单纯疱疹病毒(HSV)抗原在临床中的应用价值.方法用聚合酶链反应(PCR)和ELISA方法检测泌尿生殖道及皮损中HSV DNA和HSV抗原,并对两种方法的检测结果进行比较.结果118例标本中,PCR检出41例阳性,ELISA 42例阳性.在118例标本中,111例两种方法结果相符,7例不相符合.PCR阳性的41例中,ELISA阳性38例(敏感性92.68%);PCR阴性的77例标本中,ELISA阳性4例(特异性94.81%).结论ELISA检测HSV抗原的方法可直接检测出泌尿生殖道及皮损中HSV病原体,从而为生殖器疱疹诊断提供准确的实验依据,其敏感性、特异性高,方便,快速,值得临床推广应用.     

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。     

特异性HSV抗体检测在生殖器疱疹诊断中的重要性分析

目的探讨生殖器疱疹(GH)患者HSV-Ⅰ、Ⅱ型IgG、IgM抗体检测的临床意义,评价其对GH诊断的重要性和实用性。方法应用酶联免疫吸附试验(ELISA)对生殖器部位有皮损者检测HSV的抗原及IgG、IgM抗体。结果男性患者有生殖器皮损者215例,HSV抗原阳性率45.58%(98/215),抗体阳性率98.14%(211/215)。女性患者有生殖器皮损者193例,HSV抗原阳性率为38.86%(75/193),抗体阳性率99.48%(192/193)。生殖器有皮损者HSV抗体阳性率均明显高于抗原阳性率。男女IgMⅠ型阳性率42.37%(100/236),提示HSV-Ⅰ所致GH明显增加。结论对生殖器皮损时间短的进行HSV抗原检测,及对生殖器皮损时间长或反复者进行抗体检测是判断HSV感染的检测方法。IgM抗体检测对有生殖器皮损但抗原检测阴性,及对无皮损排毒期具有辅助诊断意义。     

单纯疱疹病毒在不典型皮损及无皮损的生殖道分泌物检测情况的临床分析

目的研究不典型皮损及无皮损生殖道分泌物单纯疱疹病毒(HSV)的检测情况,为生殖器疱疹的防治提供临床研究资料。方法回顾性分析我院性传播疾病(STD)门诊379例疑似生殖器单纯疱疹感染患者临床资料。结果典型组HSV检出86例,检出率33.33%,非典型组HSV检出17例(HSV-2型13例、HSV-1型4例),检出率14.05%,典型组HSV检出率较高,差异有统计学意义(χ2=14.479,P=0.000,P0.05);典型组不同皮损HSV-2型检出率比较,差异均无统计学意义(χ2=1.369,P=0.713,P0.05);非典型组不同皮损HSV-2型检出率比较,差异均无统计学意义(χ2=0.779,P=0.855,P0.05);男性疑似HSV感染患者HSV检出率24.73%,女性疑似HSV感染患者HSV检出率32.49%,差异有统计学意义(P0.05),女性检出率较高。结论对非典型皮损或无皮损的生殖道分泌物检测HSV病毒对于HSV病毒亚临床感染及时诊断、治疗及传播的防范均有重要的临床意义。     

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。     

单纯疱疹病毒的聚合酶链反应——微孔板反向杂交检测和分型

单纯疱疹病毒 2型 (HSV－ 2)和单纯疱疹病毒 1型 (HSV－ 1)引起的生殖器疱疹的预后不同 [1]，检测 HSV时有必要对其分型。目前，生殖器疱疹的实验室诊断“金标准”是 HSV分离培养法，该法敏感性欠佳，费时费力。新近出现的 PCR－ ELISA及 PCR－微孔板杂交法，将 PCR、核酸杂交及酶联免疫技术结合起来，克服了传统方法的诸多弊端 [2,3]。我们建立了 HSV的 PCR－微孔板反向杂交检测和分型方法，并初步评价其在生殖器标本 HSV检测和分型中的应用价值。 一、材料与方法 (一 )材料： Vero细胞自中国科学院上海细胞生物研究所引…     

荧光定量聚合酶链反应在生殖器疱疹诊断和分型中的应用

目的评价荧光定量聚合酶链反应(FQ-PCR)在生殖器疱疹(GH)诊断和分型中的应用。方法以单纯疱疹病毒(HSV)-1 DNA多聚酶基因和HSV-2糖蛋白D基因为靶基因区,设计合成正向、反向引物和探针,分别对HSV-1和HSV-2进行FQ-PCR检测,优化反应体系,进行方法学评价,并对疑似GH患者的生殖器棉拭子标本采用FQ-PCR进行检测和分型。结果建立的FQ-PCR对HSV检测和分型具有特异性;线性范围好(标准品的浓度为5×102~5×108 copies/ml,r=0.998);灵敏度达到5×102 copies/ml;重复性较好,批内CV值为2.29%,批间CV值为4.76%。186例患者HSV阳性44例,阳性率为23.7%(44/186);其中44例阳性者中HSV-1 8例(占18.2%),标本病毒载量为8.5546×106 copies/ml,HSV-2 36例(占81.8%),病毒载量为1.9 861×106 copies/ml。FQPCR与PCR法的HSV检出率与分型检测结果一致。结论建立的FQ-PCR方法具有高特异性和敏感性,分型、定量准确,方法快速、简便,可用于GH的诊断和分型。     

生殖器溃疡中单纯疱疹病毒的检测和分型

目的：了解性病门诊生殖器溃疡患者中单纯疱疹病毒（HSV）感染情况，并评价聚合酶链反应（PCR）－微孔板反向杂交检测和分型方法在生器疱疹诊断中的意义。方法：采用病毒分离培养、普通PCR和PCR－微孔板反向杂交法同时对200份生殖器溃疡标本作了HSV检测与分型。结果：PCR－微孔板反向杂交法的敏感性和特异性分别为98．1％和95．9％，PCR－微孔板杂交法分型结果与病毒分离培养法和普遍PCR的分型结果完全相符。生殖器溃疡中HSV检出率为30％（60／200），其中HSV－2感染占96．7％（58／60）。结论：HSV－2是性病门诊患者生殖器溃疡的主要病因之一，PCR－微孔板反向杂交法是一种适用生殖器溃疡标本中HSV的检测与分型的快速、敏感和特异的诊断方法。     

生殖器疱疹127例临床和实验室诊断的分析

目的探讨生殖器疱疹患者的临床特征,进行实验室检测方法的比较。方法回顾分析127例生殖器疱疹患者临床资料,并同时进行HSV抗原ELISA检测及血清学检测。结果生殖器疱疹患者好发于青年人,发病前可有诱因,皮损多样。127例经ELISA检测,阳性79例,阳性检出率为62.2%,HSVIgG,IgMELISA检测,其中IgG阳性15例,占11.8%,IgM阳性6例,占4.7%。结论生殖器疱疹皮损多样,应提高警惕。抗原ELISA法优于血清IgM,IgG抗体血清法。     

Polymerase chain reaction for diagnosis of genital herpes in a genitourinary medicine clinic

BACKGROUND: Polymerase chain reaction (PCR) has well established advantages over culture for diagnosis of herpes viruses, but its technical complexity has limited its widespread application. However, recent methodological advances have rendered PCR more applicable to routine practice. Aim: To compare automated PCR with viral culture for diagnosis of genital herpes. METHODS: We studied 236 patients presenting with clinical features suggestive of genital herpes at an inner city genitourinary medicine clinic. Two swabs were taken from each patient. Cell culture and typing were performed by standard methods. Automated PCR was performed using the LightCycler instrument and the infecting viral type was determined by restriction endonuclease digestion of amplicons. RESULTS: 109 patients (46%) had a positive test for herpes simplex virus (HSV). In 88, both PCR and culture were positive; in 21 PCR only was positive. With both detection methods, lesion duration and morphology were associated with HSV detection. Compared with culture alone, use of PCR increased sensitivity by 13.3% in specimens from vesicular lesions, by 27.4% from ulcerative lesions, and by 20.0% from crusting lesions. CONCLUSIONS: We advocate adoption of automated PCR as an efficient HSV detection and typing method for diagnosis of genital herpes in routine clinical practice. PCR allowed rapid laboratory confirmation of the diagnosis and increased the overall HSV detection rate by 24%.     

Detection of varicella zoster virus in genital specimens using a multiplex polymerase chain reaction

OBJECTIVE: To compare the relative proportions of varicella zoster virus (VZV) and herpes simplex viruses in specimens obtained from the genital lesions of adults presenting with presumed genital herpes infection. METHODS: Swabs of genital lesions from 6210 patients attending general practices, infectious diseases clinics within hospitals, or sexual health centres for treatment of their genital lesions were tested using polymerase chain reaction (PCR) technology. The multiplexed PCR was capable of detecting herpes simplex virus types 1 and 2 (HSV-1, HSV-2), VZV, and cytomegalovirus in a single sample. RESULTS: A total of 2225 patients had viruses detected by PCR. HSV-1 was detected in 36%, HSV-2 in 61%, and VZV in 2.9% of PCR positive samples. Of the 65 patients with VZV genital infection, many were thought to have HSV infection before laboratory testing. CONCLUSIONS: The finding of VZV in nearly 3% of virus positive genital specimens demonstrates that this virus needs to be considered as a differential diagnosis for genital herpetic lesions. Advice provided to patients with VZV genital infection regarding the source of infection, likelihood of recurrence, and potential for transmission of the virus will be different from that given to patients with HSV infection.     

抗原捕获聚合酶链反应分型检测妇女生殖器单纯疱疹病毒

目的 :　建立直接分型检测妇女生殖器单纯疱疹病毒 (HSV)的抗原捕获聚合酶链反应 (AC -PCR)。方法 :　用抗HSV型共同性糖蛋白单克隆抗体 ,包被聚苯乙烯离心管 ,捕获HSV ,同时加入 3个引物 :HSV - 1 HSV - 2型共同性上游引物及HSV - 1和HSV - 2型特异性下游引物 ,进行PCR扩增。结果 :HSV - 1和HSV - 2标准病毒株均分别扩增出与设计大小相符的 4 77bp和 399bpDNA条带。AC -PCR可检测到 10PFUHSV - 1和 1PFUHSV - 2。用AC -PCR检测了 36 5份妇女生殖器拭子标本 ,阳性 10 1例(2 7.7% ) ,2 3例为HSV - 1(占 2 2 .8% ) ,78例为HSV - 2 (占 77.2 % ) ;其中 112份标本同时用AC -PCR和分离培养法检测 ,AC -PCR的阳性率为 2 6 .8% (30 112 ) ,分离培养法的阳性率为 2 0 .5 % (2 3 112 ) ,两者差异有显著性 (χ2 =4 .5 ,P <0 .0 5 )。结论 :　AC -PCR是特异、敏感、快速分型检测妇女生殖器HSV感染的方法     

生殖器疱疹患者病毒DNA的定量测定研究

目的 对生殖器疱疹病毒(HSV)患者病毒DNA进行定量测定.方法 以标准的HSV质粒作为标准,用聚合酶链反应(PCR)和酶联免疫吸附法(ELISA),定量测定HSVDNA.结果 100例生殖器疱疹患者中93例HSV测定阳性,7例阴性.在93例阳性者中有58例为HSV-2(占62.4%),35例为HSV-1(占37.6%).93例阳性者定量测定结果,250μL标本混悬液中DNA质粒数为115～1.1×10⁵个,平均7.1×10⁴个;58例HSV-2阳性者,250μL标本悬液DNA质粒数为136～1.1×10⁵个,平均7.6×10⁴个;35例HSV-1阳性者DNA质粒数为115～9.4×10⁴个,平均6.3×10⁴个.分别随机取HSV-2和HSV-1阳性患者各8例已淬取和纯化的DNA混悬液10μL,定量测定结果显示:HSV-2患者最高为2.7×10⁴个DNA质粒数,最低35个,平均1.8×10⁴个.HSV-1最高2.5×10⁴个,最低29个,平均1.6×10⁴个.结论 所用几种检测法中ELISA定量总阳性率为93%,与DNA印迹法阳性率相同.诊断PCR阳性率为91%,HSV分型PCR阳性率为88%.     

First episodes of genital herpes in a Swedish STD population: a study of epidemiology and transmission by the use of herpes simplex virus (HSV) typing and specific serology

OBJECTIVES: To determine the proportion of herpes simplex virus type 1 (HSV-1) and HSV type 2 (HSV-2) in first episodes of genital herpes. To evaluate the use of HSV specific serology for classifying first episodes of genital herpes and for defining HSV serostatus in the patients' sexual partners. METHODS: 108 consecutive patients with first episodes of genital herpes seen at three STD clinics in Sweden from 1995 to 1999 were included in the study. HSV culture and typing were performed and serum was tested for antibodies against a type common HSV antigen and a type specific HSV-2 antigen, glycoprotein G2 (gG2). A structured interview including questions about sexual behaviour and sexual partners was taken. "Steady" partners were offered a blood test for HSV serology and counselling. RESULTS: Of 108 patients, 11 had a negative HSV culture. Of the 97 who were HSV culture positive, 44% (43/97) were typed as HSV-1 and 56% (54/97) as HSV-2. For 86 of these 97 patients, HSV serology from the initial visit was available. Of 52 primary infections, thus initially seronegative, 64% were HSV-1 infections and of 19 female primary infections 16 (84%) were HSV-1. In 17% the first episode of genital herpes corresponded to the first clinical recurrence of an infection acquired earlier in life. There was a significant correlation between having orogenital sex and being infected with HSV-1 and also a history of labial herpes in the partner. Only 20% of partners of patients with an HSV-2 infection had a history of genital herpes. CONCLUSIONS: Almost half of first episodes of genital herpes are caused by HSV-1. In young women with a primary genital infection, HSV-1 is much more frequent than HSV-2. Besides HSV typing, we found specific HSV serology of value for classifying first episodes and for diagnosing a subclinical HSV-2 infection in partners. Anamnestic data supported the suggestion that the orogenital route of transmission was common in genital HSV-1 infections.     

Herpes simplex virus type 1 is the prevailing cause of genital herpes in the Tel Aviv area, Israel

BACKGROUND AND GOAL: The changing epidemiology of genital herpes with the increasing importance of herpes simplex virus (HSV) type 1 prompted a study on the relative prevalence of HSV-1 and HSV-2 among cases of genital herpes in the Tel Aviv area, Israel. STUDY DESIGN: A retrospective laboratory-based study of positive genital and nongenital herpes cultures performed at the Beilinson Medical Center between 1993 and 2002. Data regarding the number of isolates of each type and the age and sex of patients with genital lesions were retrieved from the database. Cultures were performed using Vero cells, and positive results were confirmed and typed by immunofluorescence. RESULTS: A total of 285 positive genital cultures and 659 positive nongenital cultures were recorded. HSV-1 was identified in 189 (66.3%) of the positive genital specimens and in 656 (99.55%) of the nongenital specimens. HSV-1 was isolated in 174 of 262 (66.4%) female subjects and 15 of 23 (65.2%) male subjects. The proportion of HSV-1 genital isolates was 72.7% in patients 15 to 24 years of age, 62% in those 25 to 44 years, and 46% in those aged 45 years or older. Overall, the annual isolation rate of genital HSV-1 has not changed markedly over the years. CONCLUSION: Herpes simplex virus type 1 has clearly been the predominant HSV type isolated from genital specimens in the Tel Aviv area over the last decade.     

Duplex real-time polymerase chain reaction assay for detection and quantification of herpes simplex virus type 1 and herpes simplex virus type 2 in genital and cutaneous lesions

BACKGROUND: A sensitive and specific method for detecting herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) is important for diagnosing genital and cutaneous infections. GOAL: The goal of this study was to compare quantitative real-time polymerase chain reaction (qPCR) with virus culture for diagnosis of genital and cutaneous HSV-1 and HSV-2. STUDY DESIGN: A duplex qPCR system for quantification of DNA from HSV-1 and HSV-2 was developed. Duplicate swabs for PCR and virus culture were collected from 89 patients attending our sexually transmitted infection and dermatology clinic. RESULTS: The duplex qPCR had a linear measure interval of 10-10 copies/mL. The detection limit was between 1 and 5 copies per reaction. qPCR detected HSV in 57 (64%) specimens and virus was isolated in 45 (50%) cases. First-episode infections showed higher viral quantities with a median value of 4.2 x 10 copies per reaction compared with recurrent infections with 1.0 x 10 (P = 0.0002). HSV-1 was more likely to be the cause of first-episode genital infections (72%), and HSV-2 of recurrent and atypical genital manifestations (73%). CONCLUSION: Real-time PCR is a sensitive method for diagnosing genital herpes, and the duplex format is convenient for typing. The method increased the detection rate by 27% compared with virus culture.     

Rapid detection of glycoprotein G gene for the diagnosis and typing of herpes simplex virus infection in genital herpes

OBJECTIVE: To develop a new, rapid, and convenient technique for the diagnosis and typing of herpes simplex virus (HSV) in genital herpes (GH). METHODS: Using samples from skin vesicle fluid and urogenital mucosal swabs of subjects with GH, conventional polymerase chain reaction (PCR) (directed to polymerase gene: PCRpG) were compared with a newly developed PCR (directed to HSV glycoprotein gene: PCRgG). Both PCR methods were compared with virus isolation culture (VI) with indirect immunofluorescent staining (IIF). RESULTS: 80 samples from 40 GH patients (25 males) were tested. Positive results were seen in 52.5% (42/80) using PCRgG compared with 40% (32/80) by VI. Most of PCRgG positive samples (95.1%) were caused by HSV-2 infection. In samples from healing lesions, HSV was detected more often by PCRgG, than by VI. The results of typing by PCRgG and IIF were highly consistent. CONCLUSION: PCRgG is more sensitive than VI and PCRgG in detecting HSV in urogenital samples from subjects with GH. PCRgG is a convenient technique for the rapid detection and typing of GH.