Rapid detection of cutaneous herpes simplex virus infection with the polymerase chain reaction

A simple and specific method for detecting herpes simplex virus infection in routinely processed paraffin-embedded biopsy specimens is described. DNA is extracted from paraffin blocks, and subjected to DNA amplification with the polymerase chain reaction. After 40 rounds, an amplified band can be detected after agarose gel electrophoresis and ethidium bromide staining. This band is specific for herpes simplex virus, because tissues infected with related viruses do not give this amplified band. We have been able to detect viral DNA in small punch skin biopsies with this procedure, which can take as little as 6 h.     

Routine detection of herpes simplex virus and varicella zoster virus by polymerase chain reaction reveals that initial herpes zoster is frequently misdiagnosed as herpes simplex

The differential diagnosis of herpes simplex and zoster may require virological confirmation, yet virus typing is not regarded as necessary in routine dermatological assessment. In an attempt to evaluate the clinical benefits of the routine detection of herpes simplex virus (HSV) and varicella zoster virus (VZV), we analysed skin swabs from 110 patients who were diagnosed at the first clinical visit as having herpes simplex ( n  = 45) or zoster ( n  = 65). Viruses were typed using the polymerase chain reaction (PCR) with the general primer pair GPHV-RU. PCR analysis showed that at the initial clinical presentation, herpes simplex in these patients was not mistaken for zoster but that zoster was incorrectly diagnosed as herpes simplex in nine cases. Thus these results suggest that initial zoster often mimics herpes simplex, hence routine PCR diagnosis of HSV and VZV or alternative rapid diagnostic approaches may be beneficial in these cases.     

Identification of herpes simplex virus DNA in lesions of erythema multiforme by the polymerase chain reaction.

An association between erythema multiforme and herpes simplex virus infection has been supported by clinical studies and by the detection by immunofluorescence of herpes viral antigen in sera and skin biopsy specimens of patients with erythema multiforme. In rare cases, the virus has also been isolated in cultures of skin biopsy specimens of erythema multiforme. To investigate further the association between erythema multiforme and herpes simplex virus, we used the polymerase chain reaction for herpes simplex virus to examine skin lesions from patients with erythema multiforme. In this study herpes simplex virus DNA was detected in 11 of 31 biopsy specimens of erythema multiforme; six additional cases showed equivocal amplification results, which is suggestive of low amounts of viral DNA. Seven skin and mucosal biopsy specimens with the histologic changes of herpes virus infection served as positive controls: all were positive for herpes simplex virus DNA. Viral DNA was not detected in control biopsy specimens from skin excised for unrelated conditions. These studies support the association of herpes simplex virus in the pathogenesis of some cases of erythema multiforme. The polymerase chain reaction provides a quick and effective method of detecting herpes simplex virus in lesions of herpes-associated erythema multiforme. Furthermore, the polymerase chain reaction may delineate those cases of erythema multiforme that are etiologically related to herpes virus infection and therefore might be treated with acyclovir to prevent recurrence.     

Natural history of genital herpes simplex virus type 1 infection

BACKGROUND: Herpes simplex virus type 1 (HSV-1) has been increasingly reported as a cause of genital herpes, yet there have been few studies on the long-term natural history of this infection.GOAL The goal was to examine the clinical course of genital HSV-1 infection. STUDY DESIGN: This was a cohort study of patients presenting with culture-proven primary genital HSV-1 infection. RESULTS: The median follow-up of the 77 patients was 736 days. The overall rate of recurrences was 1.3/year in the first year of infection, decreasing to 0.7/year in the second year. In the first year of infection, 43% of study patients did not have a recurrence. In the second year of infection, 67% of study patients did not have a recurrence. CONCLUSION: Genital HSV-1 recurs infrequently in most patients, and the rate decreases further in the subsequent years of infection. Because the prognoses of genital HSV-1 and HSV-2 infections differ, determination of the viral type is important for patient counseling.     

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。     

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     

Investigation of herpes simplex virus DNA in pityriasis rosea by polymerase chain reaction

BACKGROUND: Pityriasis rosea (PR) is an acute, inflammatory disease of unknown cause. Clinical and experimental findings indicate an infectious etiology of PR. Our purpose is to examine the skin lesions and blood samples of PR patients by polymerase chain reaction (PCR) for the presence of HSV type 1 and 2 DNA. METHODS: The lesional skin biopsies from 10 patients and blood samples from two randomized patients with clinically and histologically confirmed pityriasis rosea were examined by PCR. RESULTS: No HSV 1 and HSV 2 DNA was detected in the lesional biopsy and blood samples. CONCLUSIONS: We could not identify a relationship between HSV 1, HSV 2 and PR.     

单纯疱疹病毒1型致原发性生殖器疱疹一例

患者女,23岁。因外阴红肿、刺痛4 d入院。4 d前出现会阴瘙痒、分泌物增多,查念珠菌阳性,拟诊念珠菌性阴道炎,给予氟康唑胶囊150 mg顿服,复方明矾散、双唑泰栓及联苯苄唑乳膏外用,3 d前瘙痒加重,出现轻微疼痛,予聚维酮碘溶液阴道冲洗,制霉菌素阴道泡腾片及硝酸咪康唑栓治疗,症状无改善,2 d前外阴明显肿胀,大量水疱及脓疱,渗出,行走时疼痛明显,外院拟急性女阴溃疡给予头孢曲松、泼尼松治疗,效果不佳,疼痛加剧,呈反复持续发作的刺痛,加用盐酸曲马多注射液肌内注射,疼痛未能改善……     

Confirmation of herpes simplex virus type 2 infections in herpes-like genital lesions by a simple complement-fixation test

The presence of complement-fixing antibody to an early herpes simplex virus type 2 (HSV-2) antigen (the AG-4 antigen) was correlated with HSV-2 infection in the sera of patients with genital herpes. Eighty-eight per cent of sera taken two weeks after clinical diagnosis of a primary or recurrent herpes infection in patients, confirmed to have HSV-2 by virus isolation and typing, contained the anti-AG-4 complement-fixing antibody. None of the patients with genital HSV-1 had the antibody, and only 9% of controls or patients with facial HSV-1 infection had positive results for the antibody. This correlation was used to identify genital HSV-2 infections when either no virus sample had been taken or when virus isolations had been unsuccessful. Thus, a simple complement-fixation test can confirm an HSV-2 virus infection without isolation of the virus from the herpetic lesion.     

Comparison of the Tzanck test and polymerase chain reaction in the diagnosis of cutaneous herpes simplex and varicella zoster virus infections

Background: Although the diagnosis of herpes simplex virus (HSV) and varicella zoster virus (VZV) infections is usually made clinically, the Tzanck test, electron microscopy, viral culture, polymerase chain reaction (PCR), and serologic tests can be utilized to verify the diagnosis. METHODS: We conducted a study on a total of 98 patients (77 patients with recurrent herpes simplex and 21 patients with herpes zoster) to evaluate the reliability and reproducibility of the Tzanck test in comparison with PCR. RESULTS: In herpes virus infections, the general positivity rates of the Tzanck test and PCR were 61.2% and 79.6%, respectively. The difference between the positivity rates of the two tests was statistically significant. The positivity rates of the tests differed according to the type and duration of the lesions. CONCLUSIONS: Although PCR was superior to the Tzanck test, the Tzanck test has also been proven to be a reliable diagnostic method, with a sensitivity of 76.9% and a specificity of 100%. We recommend the use of this easy, quick, reproducible, and inexpensive diagnostic test more often in dermatologic practice, especially in cutaneous herpes virus infections.     

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。     

Epidemiology of recurrent genital herpes simplex virus types 1 and 2

OBJECTIVES: To describe the epidemiology of type specific recurrent genital herpes, and to compare the duration of recurrent genital lesions caused by herpes simplex virus (HSV) types 1 and 2. METHODS: Participants were enrolled at clinics across the United States. Adults suspected of having active genital herpes were eligible. Lesions were cultured for HSV and typed. Data from 940 participants with recurrent culture positive HSV lesions were analysed. Pearson's chi(2) and Fisher's exact tests, multivariate logistic regression models, and a stratified Cox proportional hazards model were used to compare epidemiological characteristics and lesion duration of HSV-1 and HSV-2. RESULTS: HSV-1 was present in 4.2% of the recurrent HSV culture positive lesions. HSV-1 was most prevalent among whites (6.5%) and individuals with 0-2 recurrences in the previous year (9.1%) and, among men, in those with rectal/perirectal lesions (13.2%). Longer lesion duration was not significantly associated with virus type (hazard ratio (HR) 0.95, 95% confidence interval (CI) 0.65 to 1.38, p = 0.79), but was associated with male sex (HR 0.85, 95% CI 0.74 to 0.99, p = 0.04), and HIV seropositivity (HR 0.62, 95% CI 0.48 to 0.81, p<0.01). CONCLUSIONS: The authors found that, in the United States, recurrent genital HSV-1 is relatively rare in the STD and HIV clinic setting, especially among black people. Among men, rectal/perirectal recurrent lesions are more likely to be caused by HSV-1 than are penile lesions. In addition, lesion duration depends on sex and HIV status but not virus type. These findings shed new light on the type specific epidemiology of recurrent genital HSV, and suggest that type specific testing can inform the prognosis and management of genital herpes.     

单纯疱疹病毒抗原检测在生殖器疱疹诊断中的临床评价

生殖器疱疹(genital herpes,GH)是由单纯疱疹病毒(herpes simplex virus,HSV)感染引起的一种常见的性传播疾病。临床表现多种多样,是生殖器溃疡最     

Confirmation of herpes simplex virus type 2 infections in herpes-like genital lesions by a simple complement-fixation test.

The presence of complement-fixing antibody to an early herpes simplex virus type 2 (HSV-2) antigen (the AG-4 antigen) was correlated with HSV-2 infection in the sera of patients with genital herpes. Eighty-eight per cent of sera taken two weeks after clinical diagnosis of a primary or recurrent herpes infection in patients, confirmed to have HSV-2 by virus isolation and typing, contained the anti-AG-4 complement-fixing antibody. None of the patients with genital HSV-1 had the antibody, and only 9% of controls or patients with facial HSV-1 infection had positive results for the antibody. This correlation was used to identify genital HSV-2 infections when either no virus sample had been taken or when virus isolations had been unsuccessful. Thus, a simple complement-fixation test can confirm an HSV-2 virus infection without isolation of the virus from the herpetic lesion.     

Acquisition of concomitant oral and genital infection with herpes simplex virus type 2

Primary oral and genital infections caused by herpes simplex virus type 2 were diagnosed in an 18-year-old female. A history of sexual practices was critical in determining the anatomic sites of infection. Restriction endonuclease analysis of viral DNAs helped to identify the male sexual partner from whom the virus had been acquired. He had been infected recently by a previous sexual partner but had not yet developed lesions. Clinicians should obtain a history of sexual practices from patients with newly acquired genital herpes and should advise patients with genital herpes that transmission of virus to sexual partners can occur in the absence of overt lesions.     

酶联免疫吸附试验检测单纯疱疹病毒抗体在生殖器疱疹筛查中的应用

目的:探讨酶联免疫吸附试验检测单纯疱疹病毒(HSV)抗体对生殖器疱疹筛查的意义。方法:选取50例2010年2月至2013年10月来我院治疗疑似生殖器疱疹患者,将其作为试验组,使用酶联免疫吸附试验进行HSV抗体检测,另外选择50例健康体检人员血清作为对照组。结果:在50例疑似生殖器疱疹患者HSV-I Ig M,HSV-I Ig G的检出率依次为14%,32%,HSV-ⅡIg M检出率为20%,HSV-ⅡIg G的检出率为34%;在对照组中HSV-I Ig M,HSV-I Ig G的检出率依次为0.8%,14%,HSV-ⅡIg M检出率为10%,HSV-ⅡIg G的检出率为24%,两组患者HSV抗体检测阳性率有显著的不同,二者明显的差异具有统计学意义(P0.05)。结论:疑似生殖器疱疹患者单纯疱疹病毒抗体检出率高,酶联免疫检测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,在生殖器疱疹筛查中具有一定的优势。     

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.     

Sequential genital infections with herpes simplex virus types 1 and 2.

Herpes simplex virus (HSV) types 1 and 2, typed by an enzyme linked immunosorbent assay (ELISA), were isolated at different clinical episodes from five people with genital herpes. This finding has important implications for assessing resistance to antiviral drugs in therapeutic studies.     

Proportion of herpes simplex virus (HSV) type 1 and type 2 among genital and extragenital HSV isolates

Herpes simplex virus type 1 (HSV-1) has been associated with orofacial infections and HSV type 2 (HSV-2) with genital infections. This tropism of the virus seems to have changed and in clinical reports an increasing number of genital herpes infections caused by HSV-1 have been recognized. The aim of this study was to estimate the proportion of HSV-1 and HSV-2, respectively, among isolates from different anatomical sites typed in our laboratory during the years 1994-1998. Out of a total of 3,085 anogenital isolates, 29% were typed as HSV-1 and 71% as HSV-2. The highest prevalence of HSV-1 was registered among isolates from young women. Of 631 orofacial isolates, 4% were typed as HSV-2 and 96% as HSV-1. Of 69 finger/hand isolates, 54% were typed as HSV-1 and 46% as HSV-2, and of 95 isolates from other regions (abdomen, foot, etc.), 60% were typed as HSV-1 and 40% as HSV-2. It was found that HSV-2 was as common as HSV-1 in the extra-genital regions with the exception of the orofacial area, in which HSV-2 was seldom detected. Furthermore, the study showed an increasing proportion of HSV-1 among anogenital isolates during the study period. Taken together, these results suggest that a clear HSV type-related tropism might be limited to the permissiveness of the orofacial region for HSV-1, and that both serotypes may readily establish infections below the neck.