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。     

生殖器疱疹病毒感染检测方法的临床应用

目的 :　评价检测生殖器疱疹病毒实验方法的临床应用价值。方法 :　同时采用细胞培养法、定量PCR法、间接免疫荧光法 (IIF)对 5 3例生殖器疱疹 (GH)感染患者进行了HSV检测。结果 :　细胞培养、定量PCR检测HSV阳性率明显高于IIF法 (P <0 .0 1) ,细胞培养与定量PCR比较无显著性差异 (P >0 .0 5 )。三种方法检测GH患者皮疹水疱内的阳性率无明显差异 (P >0 .0 5 )。IIF法检测糜烂结痂的阳性率仅为 2 8.6 %。结论 :　三种方法均适用GH水疱期患者标本的检查。而定量PCR、细胞培养检测GH糜烂结痂患者标本较IIF法敏感     

间接免疫荧光法检测生殖器疱疹病毒

目的:评价间接免疫荧光试验(IFA)在生殖器疱疹(GH)诊断中的应用价值。方法:采用以单纯疱疹病毒(HSV)型共同性单克隆抗体为夹心的IFA法,检测了120例临床诊断为GH患者皮疹中的HSV,并与病毒培养法进行比较。结果:IFA检测HSV的总阳性率为85.8%,高于病毒培养法的阳性率(70.8%,χ2=12.04,P<0.01)。两种方法检测GH水疱内的HSV阳性率分别为93.3%和90.0%,无明显差异(χ2=1.96,P>0.05);而检测糜烂和结痂性皮疹内的HSV时,IFA的阳性率分别为92.6%和69.4%,均分别高于病毒培养法(75.9%,χ2=5.82,P<0.05;47.2%,χ2=14.17,P<0.01)。结论:IFA法具有简单、快速、敏感性高的优点,适于检测GH患者皮疹内HSV,有临床实用价值。     

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。     

性传播疾病

20 0 12 70 0　性传播疾病临床研究进展 (综述 ) /杨森 (安徽医大附院皮肤科 )∥安徽医学 .- 2 0 0 0 ,2 1( 6) .- 64～662 0 0 12 70 1　性病患者 H SV- 2抗体检测研究 /谭志建(武汉同济医大协和医院皮肤科 )…∥临床皮肤科杂志 .- 2 0 0 1,30 ( 1) .- 2 5对 2 97例不同性病患者、30名健康者进行血清 2型单纯疱疹病毒 ( HSV - 2 )的抗体检测。结果各种性病患者均有不同比例的 HSV- 2感染 ,以梅毒患者阳性率最高。H SV- 2抗体阳性可提示潜在感染的可能。有报道 H SV- 2抗体阳性者 ,60 %为未识别症状的生殖器疱疹 ( GH )患者 ,如生殖…     

对ELISA检测生殖器疱疹患者HSV及其临床应用的评价

目的评价酶联免疫吸附试验(ELISA)检测生殖器疱疹病毒的临床应用价值。方法采用ELISA和分型聚合酶链反应(分型PCR)检测生殖器标本中的单纯疱疹病毒(HSV),两种试验结果不符合者采用不分型PCR检测。结果164例受检者中,ELISA法HSV阳性96例(58.5%),其中具典型皮损者阳性84例(80.8%,84/104),非典型皮损阳性12例(20.0%,12/60);分型PCRHSV阳性98例(59.8%),其中典型皮损者HSV阳性86例(82.7%,86/104),非典型皮损者阳性12例(20.0%,12/60)。HSV1感染者占生殖器疱疹的5.1%,HSV2感染占88.7%,HSV1和HSV2混合感染者占6.1%。ELISA的敏感性和特异性分别为96.7%和94.0%。结论ELISA检测HSV感染,其敏感性高、特异性强,方便、快速,尤其适合大批量样本的检测。     

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。     

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。     

荧光多重实时PCR检测单纯疱疹病毒

目的建立单纯疱疹病毒的荧光多重实时PCR检测及分型技术，为生殖器疱疹的诊断及病毒分型提供快捷、敏感、特异的方法。方法以细胞培养法为对照，用Smart Cycler System作为实验平台，以DNA探针荧光标记技术建立荧光多重实时PCR，用于检测生殖器疱疹患者临床标本中的1型和2型单纯疱疹病毒(HSV1、HSV2)DNA。结果荧光多重实时PXR检测HSV1和HSV2 DNA的敏感性和特异性均为100％，可检测出低至1～5拷贝的HSV DNA，30min左右即可完成整个检测过程．而细胞培养的敏感性和特异性分别为70．10％和100％。结论基于Smart Cycler System的荧光多重实时PCR是一种易于操作、扩增效率高、封闭式的核酸扩增技术，可用一次PCR反应就能快速、敏感、准确地对临床标本中的单纯疱疹病毒进行检测和分型。     

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。     

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.     

荧光多重PCR与血清型特异性抗体检测HSV感染的比较

目的：比较荧光多重PCR和血清型特异性抗体测定在生殖器疱疹临床诊断中的应用价值及评价各自的优缺点。方法：以细胞培养法作为“金标准”对照，分别用荧光多重PCR和血清型特异性抗体检测法对121例临床诊断为生殖器疱疹的标本进行检测。结果：以培养法作标准，并通过结果的差异性分析，荧光多重PCR的敏感性为100％，特异性为88．89％；血清型特异性抗体测定则分别为77．68％和77．78％，荧光多重PCR的敏感性和特异性均显高于血清型特异性抗体测（P＜0．05），但前不能检测出无皮损患HSV的DNA，而后可检测出无皮损患中的HSV抗体。结论：荧光多重PCR和血清型特异性抗体检测各有其自身的优缺点，单独用PCR和其它病毒分离的方法和单独使用血清特异性抗体检测的方法来诊断生殖器疱疹都是不完整的，均可造成漏诊。临床上将两种方法有机的结合起来应用能发挥各自的优势，取长补短，对早期、准确、快速地诊断生殖器疱疹及进行流行病学调查有着十分重要的意义和使用价值。     

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.     

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。     

Famciclovir reduces viral mucosal shedding in HSV-seropositive persons

OBJECTIVE: Many cases of herpes simplex virus (HSV) infection occur through asymptomatic shedding from persons without evidence of clinical disease. This study explores whether famciclovir reduces HSV shedding in HSV-2 seropositive persons with or without a history of symptomatic genital herpes. STUDY DESIGN: One hundred twenty-seven HSV-2 seropositive participants were randomly assigned to 42 days of famciclovir, followed by 14 days of washout and 42 days of placebo, or vice versa. All subjects swabbed the genital/perianal area; those with HSV-1 infection also swabbed the oral area daily for HSV DNA PCR. RESULTS: Famciclovir reduced genital and oral HSV shedding from 11.4% of days during the placebo period to 4.7% of days during famciclovir therapy. The reduction was greater in participants with a history of genital herpes (74%) than in those without such a history (30%). In multivariate analyses, famciclovir protected against total (clinical and subclinical) genital shedding among persons with a clinical history of genital herpes (RR, 0.23; 95% CI, 0.15-0.35; P < 0.001). Among HSV-2 seropositive participants without a history of genital herpes, 60% had HSV detected in the genital area at least once during the study. Famciclovir therapy did not result in a statistically significant reduction in total HSV shedding in participants without a history of genital herpes. CONCLUSION: Famciclovir therapy decreases genital HSV shedding in HSV-seropositive persons, especially those with a history of genital herpes. Overall, antiviral drugs may have varying effects on symptomatic and asymptomatic viral shedding, depending on the clinical history of the disease.     

生殖器疱疹患者尿道和宫颈HSV感染的研究

目的：了解生殖器疱疹（GH）患者伴发宫颈和男性尿道单纯疱疹病毒（HSV）感染情况，进一步探讨HSV和非淋菌性尿道炎／官颈炎（NGU）的关系。方法：用HSV聚合酶链反应（PCR）方法检测了56例GH患者的宫颈和男性尿道拭子标本。结果：56例患者中HSV PCR检测阳性共5例，阳性率为8．93％（5／56）。20例女性生殖器疱疹患者宫颈HSV PCR检测阳性率为25．0％（5／20）；36例男性GH患者尿道HSV PCR检测结果均为阴性。结论：生殖器疱疹患者伴发女性宫颈HSV感染更为常见，而伴发男性尿道HSV感染相对少见。HSV感染可能是非淋菌性尿道炎／宫颈炎的一个致病因子，在女性宫颈炎的发病中可能更有意义。     

Seroprevalence of herpes simplex virus type 1 and type 2 in Turkey

BACKGROUND: Herpes simplex virus (HSV) infections are among the most common infectious diseases in humans. The prevalence of herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) varies widely across the world. HSV-2 infection is the primary cause of genital herpes. It is highly prevalent in human populations in many parts of the world, and is the most common cause of genital ulcer disease worldwide. In spite of the large prevalence and growing incidence of herpes simplex infection (HSV-1 and HSV-2), relatively few data have been published regarding the seroprevalence of herpes simplex infection, while no data exist regarding the Turkish population. METHODS: We aimed to investigate the prevalence of HSV-1 and HSV-2 in selected populations in Turkey. A cross-sectional study was conducted involving 2082 serum samples of 725 adults, 300 pregnant women, 200 blood donors, 483 sex workers and 110 patients with genital warts and 264 hotel staff in Istanbul, Turkey. All serum samples were assessed for HSV1 and HSV-2 IgG antibodies using an HSV-type specific, enzyme-linked immunosorbent assay (ELISA). RESULTS: The prevalence of HSV-2 and HSV-1 antibodies was 4.8 and 85.3% in sexually active adults; 5.5 and 96% in blood donors; 5 and 98% in pregnant women, 17.3 and 93.6% in patients with genital warts; 8.3 and 97.3% in hotel staff; and 60% and 99% in sex workers. CONCLUSION: These results confirm a higher prevalence of HSV infection than estimated, especially in high risk groups in Turkey. The high prevalence of HSV infection underlines the need for education among these populations.     

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%.