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。     

沙眼衣原体酶免疫法和细胞培养法的比较

目的　评价一种改进的沙眼衣原体酶免疫法 (ELISA)诊断沙眼衣原体感染的临床应用价值。方法　 2 0 0例性病门诊有泌尿生殖道症状的患者采用ELISA和细胞培养法检测泌尿生殖道标本中的沙眼衣原体 ,两种试验结果不符合者采用PCR检测。结果　 2 0 0例受检者中 ,ELISA阳性 19例 ,细胞培养阳性 2 0例。与“扩大金标准”比较 ,ELISA法和细胞培养的敏感性分别为 82 .68%和 68.96% ,特异性均为 10 0 % ,阳性预期值均为 10 0 %。结论　ELISA法检测泌尿生殖道沙眼衣原体感染 ,其敏感性和特异性高 ,且方便、快速 ,尤其适合大批量样本的检测 ,值得在临床应用。     

性病门诊患者泌尿生殖道分泌物单纯疱疹病毒的检测及临床特点分析

目的:探讨性病门诊患者泌尿生殖道单纯疱疹病毒(HSV)感染的临床特点。方法:应用酶联免疫吸附试验(ELISA)对性病门诊患者泌尿生殖道分泌物及周围病损渗出液和疱液进行HSV检测。结果:1848例标本应用ELISA法检测HSV。1077例男性尿道分泌物中,伴生殖器疱疹(GH)者(47.9%,227/474)HSV阳性率高于不伴GH者(27.2%,55/202);771例女性中伴GH者宫颈分泌物(34.8%,103/296)HSV阳性率高于不伴GH者(20.7%,76/367)。男性尿道及女性宫颈分泌物HSV阳性率均低于病损渗出液和疱液阳性率。病损渗出液和疱液不同病期HSV阳性率对比,病期<3d者明显高于病期≥4d者。泌尿生殖道分泌物HSV阳性78例,同时并发其他性病病原体阳性312例(39.7%)。结论:在性病患者中部分无GH者的泌尿生殖道分泌物中可检测到HSV感染,且可并发其他性病病原体感染,对无GH的性病患者进行HSV检测是十分必要的。     

三种方法检测泌尿生殖道沙眼衣原体的比较

目的用三种方法检测泌尿生殖道沙眼衣原体,评价其方法的敏感性和特异性.方法采用C-C快速法、PCR和VIDAS CHL三种方法检测150例尿道或宫颈分泌物标本.结果在150例患者中,阳性患者43例,阳性率为28.67％,C-C快速法检测的敏感性与特异性为97.67％和100％,PCR检测的敏感性和特异性为100％和98.13％,VIDAS CHL检测的敏感性和特异性为97.67％和97.20％.结论三种方法检测泌尿生殖道沙眼衣原体的敏感性和特异性无显著性差异(P＞0.05).     

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。     

山东地区泌尿生殖道感染患者生殖支原体感染现状调查

了解山东地区泌尿生殖道感染患者生殖支原体(Mg)感染现状.采用PCR技术检测352例山东地区性病门诊患者生殖道分泌物标本的Mg,并与101名健康对照组比较.352例患者生殖道分泌物标本中Mg阳性58例(16.5%),其中41例同时合并其他病原体感染;101名健康对照组生殖道分泌物标本中Mg阳性2例(2.0%),二组阳性率差异有统计学意义(P＜0.001).山东地区泌尿生殖道感染患者有一定的生殖支原体阳性率,生殖支原体常与其他病原体感染同时存在.     

反向斑点杂交技术快速鉴定泌尿生殖道致病性支原体的研究

目的 建立PCR反向斑点杂交方法(PCR-RDB)快速鉴定泌尿生殖道常见致病性支原体.方法 以4种支原体[微小脲原体(Up)、解脲脲原体(Uu)、生殖支原体(Mg)、人型支原体(Mh)]的16SrRNA基因为靶序列设计通用引物和探针,将4种特异的寡核苷酸探针固定于尼龙膜上.利用巢式PCR扩增Up、Uu、Mg和Mh,扩增产物变性后与各特异性探针进行杂交、显色并分析结果,并对检测体系的敏感性、特异性进行测试.同时检测分析60例临床拭子标本.结果 4种特异性探针只与相应的支原体DNA杂交,与其他病原体无交叉反应,其敏感性是1 CFU.60例临床拭子标本PCR-RDB方法共筛选出支原体阳性19例,其中3例为支原体混合感染的标本(Up+Uu 2例,Uu+Mg 1例).结论 该方法可快速、敏感、准确地鉴定引起泌尿生殖道感染的致病性支原体.     

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。     

聚合酶链反应与血清酶联免疫吸附试验在生殖器疱疹中的应用对比分析

目的:考察聚合酶链反应(PCR)与血清酶联免疫吸附试验(ELISA)在生殖器疱疹中的应用效果差异。方法:选取140例生殖器皮肤、黏膜损伤患者,分别采用聚合酶链反应与血清酶联免疫吸附试验检测标本中单纯疱疹病毒,对两种结果不符患者采用第二种PCR进行检测。结果:140例样本中,PCR检测出阳性样本59例,单纯HSV-1感染患者6例,单纯HSV-2感染患者46例,混合感染患者7例。ELISA检出阳性样本57例。140例样本中,有17例结果不符,采用第二种PCR进行检查证实,ELISA检测法敏感性为96.3%、特异性为98.8%、阳性预测值为89.8%。PCR法敏感性为97.1%、特异性为92.8%、阳性预测值为95.6%。结论:相比于PCR法,ELISA法具有更高的敏感度和特异性,避免了样本间的相互污染,具有临床应用价值。     

泌尿生殖道炎细小脲原体PCR检测结果临床研究

目的探讨细小脲原体(Up)在泌尿生殖道炎中的临床特点。方法对1 477例泌尿生殖道分泌物进行Up、沙眼衣原体(Ct)、人乳头状瘤病毒(HPV)、单纯疱疹病毒(HSV)和淋球菌PCR检测和解脲脲原体(Uu)和人型支原体(Mh)培养。1 240例分泌物进行了涂片革兰染色。结果 1 477例泌尿生殖道分泌物检测病原体阳性854例,检出率为57.82%(854/1 477)。Up阳性占26.70%(228/854),其中占男性17.53%(64/365),占女性33.54%(164/489),男女Up阳性率差异有统计学意义。随着分泌物涂片革兰染色分级的增加,泌尿生殖道病原体阳性率也增加,且对比差异有统计学意义,说明分泌物涂片革兰染色对临床感染程度有一定的提示作用。结论 Up是泌尿生殖道炎主要病原体之一。Up感染可见于病人体弱者,在感染病原体量大、感染时间长等条件下可引起临床症状。PCR法检测Up对泌尿生殖道感染高危人群及有临床症状者能够做到诊断准确、快速,有临床应用价值。     

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.     

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     

Diagnosis of genital herpes by real time PCR in routine clinical practice

BACKGROUND: Virus isolation in cell culture is the recognised diagnostic gold standard for genital herpes. Although increasing evidence indicates that polymerase chain reaction (PCR) provides a more rapid and sensitive diagnostic method, its implementation in routine diagnostic settings has been limited by concerns over contamination and cost. OBJECTIVE: To evaluate the feasibility of replacing virus culture with PCR for the diagnosis of genital herpes in settings serving large populations of genitourinary medicine (GUM) attendees. METHODS: Genital swabs collected from 233 consecutive GUM attendees with suspected genital herpes were tested in parallel by virus culture and automated real time PCR. Three specimen preparation methods were evaluated and the assay reliability was assessed by repeat testing, comparison with a commercially available assay, and herpes simplex virus (HSV) sequence analysis. Probe melting temperatures (Tm) were used to differentiate between HSV types without additional post-PCR steps. RESULTS: HSV was detected in 79/233 (34%) samples by virus culture and 132/233 (57%) samples by PCR. PCR significantly increased HSV detection in both early (< 5 days) and late (> or = 5 days) presentations and in both first and recurrent episodes. HSV detection and typing by PCR was achieved within less than 4 hours leading to a significant reduction in labour compared to virus culture. Most specimens (120/132, 91%) were typed as HSV-2. Results were highly reproducible. CONCLUSIONS: Real time PCR is a highly reproducible, rapid, and labour efficient method for HSV detection in genital swabs. Its implementation is feasible in routine diagnostic settings.     

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。     

Detection of herpes simplex virus DNA in the peripheral blood during acute recurrent herpes labialis.

BACKGROUND: Although herpes simplex virus (HSV) has been detected in the peripheral blood of immunocompromised patients and in neonates with disseminated disease, the extent to which this virus may be present in the blood during a localized infection in otherwise healthy adults is unknown. OBJECTIVE: The purpose of this study was to determine whether HSV may be detected in the peripheral blood during acute recurrent herpes labialis. METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from otherwise healthy adults with recurrent herpes labialis, both during an acute episode and several weeks after the lesions had healed. The PBMCs were examined for the presence of HSV with the polymerase chain reaction (PCR) and viral culture. RESULTS: By PCR, HSV DNA was detected in 7 of 34 specimens from an acute episode but in none of 24 specimens in the convalescent stage (p less than 0.004). PBMCs from seven donors, who were seronegative for HSV, were also negative for HSV by PCR. Viral cultures of 22 PBMC specimens were negative (including four specimens that were positive by PCR). CONCLUSION: The presence of HSV DNA in the blood is a transient phenomenon limited to the period of active infection in a minority of patients with herpes labialis, although it may be important in the development of disseminated disease as well as in the pathogenesis of herpes-associated cutaneous processes such as erythema multiforme.     

Detection of cutaneous herpes simplex virus infections by immunofluorescence vs. PCR

BACKGROUND: Detection of cutaneous infections with herpes simplex virus (HSV) has proven difficult, as serum antibody tests sometimes are not sensitive and specific enough for that purpose. OBJECTIVE: This study was conducted to compare the sensitivity for detection of HSV of an immunofluorescence method (Syva Microtrak) and an internally controlled PCR. METHODS: Cutaneous swabs from skin lesions were analysed by immunofluorescence separately for HSV types 1 and 2 and by competitive PCR. Detection of PCR products was done by ELISA, if positive additionally by agarose gel electrophoresis. RESULTS: Of 79 samples 34 were PCR-positive by ELISA (34 = 100%), of which 23 (68%) were also positive on the agarose gel. Eleven samples (32%) were positive by immunofluorescence. No sample was positive by immunofluorescence and negative by PCR. CONCLUSIONS: These results demonstrate that immunofluorescence using Syva Microtrak is not suitable for exclusion of herpes simplex virus infection as sensitivity was only 32%. However, as immunofluorescence is cheaper and faster than PCR, first screening can be done with immunofluorescence, and negative samples can be investigated by PCR to finally prove or exclude the presence of HSV DNA.     

Rapid amplified enzyme linked immunosorbent assay evaluated for detecting herpes simplex virus.

A new amplified enzyme linked immunosorbent assay (amplified ELISA) kit for detecting herpes simplex virus (HSV) antigen was evaluated. Duplicate swabs were taken from 180 patients with clinically suspected herpes lesions. Tests were performed on a direct swab extract and viral transport medium containing a swab. Of the 93 culture positive specimens, 78 of the extracted samples (sensitivity 83.9%) and 72 of the swabs in transport medium (sensitivity 77.4%) were positive by amplified ELISA. A higher sensitivity (49/54, 90.7%) was obtained when the extracted swab was taken first. In early lesions the sensitivity was 93.8% but in late lesions it was 73.3%. This ELISA therefore offers an alternative to culture for early lesions, but culture is the method of choice for differential diagnosis of genital ulceration. As the specificity was 94.3%, this test is acceptable for testing populations with a high prevalence of HSV infection, but culture should be used for screening populations in which the disease is rare.     

gD基因在生殖器疱疹PCR诊断中的研究

依据国外已发表的疱疹病毒（ＨＳＶ）糖蛋白Ｄ基因（ｇＤ基因）的核苷酸保守区序列设计了一对引物，建立了用疱疹病毒ｇＤ基因做疱疹病毒诊断基因的ＰＣＲ方法。对４８例拟诊为疱疹病毒感染的临床标本进行病毒细胞培养和ＰＣＲ检测。结果表明，ＰＣＲ对ＨＳＶ２感染的敏感性为８４％（２０／２４），特异性为９１％（２２／２４）。经标准株及限制性内切酶的酶切证实和序列分析，表明该方法特异、敏感