应用McAb-ELISA双抗体夹心法分型检测脑脊液中单纯疱疹病毒抗原

神经系统的单纯疱疹病毒(HSV)感染并非少见,但常因缺乏特征性的临床表现和准确的早期病原学诊断方法,而易导致误诊、漏诊和最佳治疗时机的延误。因此,建立早期快速简便和特异性高的诊断方法就显得尤为重要。 我们应用第四军医大学分子病毒学研究窒提供的试剂盒,采用McAb-ELISA双抗体夹心法,对400例神经系统疾病患者的脑脊液(CSF)进行了单纯疱疹病毒抗原(HSV-Ag)的检测及分型。共检出15例HSV-Ag阳性患者,总阳性率为3.75％,计HSV-1     

国产化学发光法评估围孕期妇女TORCH感染情况及其IgM假阳性结果的分析

目的 探讨苏州地区围孕期女性TORCH感染情况及流行特点,并对TORCH-IgM假阳性检测结果进行分析,为本地区围孕期女性保健提供参考依据.方法 回顾性分析、统计2018年6月至2020年9月在南京医科大学附属苏州医院就诊的17984例围孕期女性TORCH检查结果,利用国产化学发光免疫分析法(CLIA)对TORCH的IgG以及IgM抗体进行检测,分析感染率.利用ELISA法对129例TORCH-IgM和IgG抗体双阳性血清进行IgG亲和力检测,分析IgM抗体假阳性结果.结果 17984例TORCH检测结果中,IgG抗体阳性率由高到低依次为CMV(97.58％)、HSV-1(89.54％)、RV(78.81％)、HSV-2(10.88％)和TOX(2.83％).IgM抗体阳性率由高到低依次为CMV(1.22％)、HSV-1(1.18％)、HSV-2(0.77％)、RV(0.57％)和TOX(0.36％),TOX感染阳性率最低,CMV感染阳性率最高.RV、CMV和HSV-1感染模式以IgM-/IgG+为主,TOX和HSV-2主要表现为IgM-/IgG-模式.按照季节统计,HSV-1-IgM在冬季阳性率最高(χ2值8.36,P值0.04),TOX-IgM、RV-IgM、CMV-IgM和HSV-2-IgM各季节阳性率差异无统计学意义(χ2值分别为2.51、4.16、1.84和5.12,P值分别为0.48、0.25、0.61和0.16).TORCH-IgG抗体阳性率四季差异均无统计学意义.按照年龄分组统计,<35岁组CMV-IgG、HSV-1-IgG和HSV-2-IgG阳性率明显低于≥35岁组(χ2值分别为12.34、15.04和238.36,均P<0.01),低年龄组TOX-IgG感染率明显高于高年龄组(χ2值为10.01,P<0.01),两年龄组间RV-IgG和TORCH-IgM抗体阳性率差异无统计学意义.化学发光免疫分析法检测129例TORCH-IgM和IgG抗体双阳性标本中,仅19例(14.73％)检测结果为IgG低亲和力.结论 苏州地区围孕期女性TORCH感染普遍,建议育龄女性孕前进行常规筛查,防止TORCH近期感染,预防出生缺陷.对于TORCH-IgM和IgG抗体双阳性结果可进一步联合IgG亲和力检测,排除TORCH感染假阳性.     

孕早期妇女TORCH感染的检测

目的 了解孕早期妇女TORCH的感染情况，以指导育龄妇女如何预防TORCH感染。方法 采用ELISA法对门诊3590例，孕早期妇女进行TORCH特异性IgM抗体检测。结果 孕早期妇女TORCH感染阳性率为：RV IgM2．53％、CMV IgM1．34％、TOX IgM0．98％、HSV-Ⅱ IgM0．53％。结论 10～12月份为本市孕早期妇女TORCH感染的高发季节，提示育龄妇女应计划受孕，做好这一时期的预防保健，进一步提高优生优育工作的质量。     

ELISA检测妇女生殖器疱疹中单纯疱疹病毒型特异性抗原的研究

用抗HSV型共同性McAb和抗HSV-2型特异性McAb包被微板,建立了能检测妇女生殖器疤疹的宫颈或阴道棉拭标本中HSV抗原,并可分型的ELISA。经与病毒分离和中和试验分型对照研究,证明本法快速、敏感、特异,分型准确可靠,可用于大规模现场普查。用本法分型检测了宫颈糜烂、霉菌性阴道炎、淋病和滴虫性阴道炎标本中的HSV抗原,结果提示上述疾病均可感染或并发感染HSV,其中以HSV-2感染为主,占24.51％,HSV-1感染次之,占6.94％。宫颈糜烂的程度与HSV阳性检出率呈正相关。淋病患者并发HSV感染的机会更多,其中HSV-1感染较其它疾病多见。故预防HSV性传播和产道感染给新生儿,应得到高度重视。     

McAb-ELISA法检测单纯疱疹病毒抗原

单纯疱疹病毒(HSV)分为HSV-1型和HSV-2型,以密切接触为主要传播途径,引起多种疱疹性疾病。其中HSV-2型是引起妇女宫颈糜烂、阴道炎的主要病原之一。本文应用McAb-ELISA法对兰州地区107例患者进行HSV-1和HSV-2型特     

HSV—2及其所致生殖道疱疹

近年来性传播疾病 ( sexually transmitteddisease,STD)日益增多 ,在 STD患者中 ,三分之二为 2 5岁及以下的青年 ,而在 STD新发病患者中 ,占 2 5%是十几岁的青少年 [1] 。单纯疱疹病毒 ( herpes simplex virus,HSV)感染 ,特别是 HSV-2感染所致的生殖道疱疹 ( genitalherpes,GH)亦呈增多趋势 ,已成为全世界范围内第二位高发的 STD[2 ] 。 GH的高发主要系无症状带毒者或未被确诊的感染者引起传播。目前认为约占 80 %的 GH病人并没有意识到自己患病。 HSV-2感染所致的疱疹应定为慢性感染性疾病 ,可持续充当传染源 ,而不是间歇性疾病…     

我国不同人群单纯疱疹病毒2型感染率及危险因素Meta分析

目的 了解我国不同人群单纯疱疹病毒2型(HSV-2)感染率水平及其危险因素.方法 采用Statal2.0对我国不同人群HSV-2感染率及相关危险因素数据进行meta分析、应用SPSS22.0进行亚组间统计学检验.结果 普通人群HSV-2感染率为16.22％ (95％ CI:12.87％ ～ 19.91％),女性高于男性,分别为16.89％(95％ CI:12.91％～21.32％)和11.69％ (95％ CI:9.07％ ～ 14.56％).不同人群既往感染率各不相同,以女性性工作者(FSW) 44.77％ (95％ CI:38.47％ ～ 46.36％)较高.多数高危人群感染率高于普通人群.随着时间推移,普通人群既往和近期感染率呈下降趋势,高危人群既往HSV-2感染率先升后降.影响因素分析提示性别、梅毒感染、HIV阳性等是感染HSV-2的危险因素.结论 我国不同人群HSV-2感染率较高,以女性、高危人群和疑与HSV-2感染相关疾病患者更为突出.建议将HIV干预和包括HSV-2在内的性病相结合、加强HSV-2监测、加强育龄妇女及孕妇HSV-2检测控制HSV-2传播.     

2014年北京地区成人呼吸道感染常见病原体IgM抗体检测分析

目的 探讨北京地区成人急性呼吸道感染(ARI)患者9种常见病毒的感染情况,分析病原体流行病学特征.方法 采用ELISA法检测1887例ARI患者巨细胞病毒(CMV)、EB病毒(EBV)、柯萨奇病毒(CVB),单纯疱疹病毒Ⅰ型和Ⅱ型(HSV-Ⅰ和HSV-Ⅱ),腺病毒3、7和11型(ADV3、ADV7和ADV11)及风疹病毒(RV)等9种病原体IgM抗体.结果 1887例标本中共检出阳性706例,总感染率为37.41％(706/1887),其中CMV感染率最高为10.92％,其次为HSV-Ⅰ(9.59％),其它病原体检出率由高到低依次为ADV7、ADV11、EBV、ADV3、HSV-Ⅱ、CVB和RV.混合感染者有260例,占36.83％ (260/706).10月份感染率最高,4月份感染率最低,且各病原体的流行季节也不同.病例以中青年感染者居多,不同性别间感染率差异无显著性.结论 2014年北京地区存在多种病原体流行,以CMV、HSV-Ⅰ和ADV7为主要病原体,各病原体流行有季节性特点.     

McAb-ELISA双夹心法分型检测单纯疱疹病毒抗原(简报)

单纯疱疹病毒(HSV)分为两型,HSV-1和HSV-2,可引起龈口炎 口唇疱疹 皮肤疱疹 疱疹性角膜炎 生殖器疱疹 疱疹性脑炎 宫颈炎等,并认为与宫颈癌的发生有关。近年来,在性传播性疾病(STD) 妇幼保健和优生优育方面,HSV的危害愈来愈受到高度重视,为了更有效控制HSV感染的流行,本文建立了一种可快速 特异检测HSV抗原的双McAb-ELISA夹心法。     

我国六省市(区)部分人群丙型肝炎病毒感染调查

目的 了解我国健康人群丙型肝炎病毒感染现状.方法 采用北京万泰生物药业的HCV EIA诊断试剂检测人群血清丙型肝炎病毒(丙肝,HCV)IgG抗体.结果 六省市(区)人群血清共9 538份,HCV抗体阳性数37份,总阳性率为0.39％,其中北京人群阳性率0.23％,黑龙江人群为0.74％,山东人群为0.26％,宁夏人群为0.10％,甘肃和四川人群阳性率均为0.44％.对37例HCV阳性者性别及年龄分析,男性19例占51.35％,女性18例占48.65％,＜10岁年龄组1例,占2.70％,10～19岁组5例,占13.51％,20～29岁4例占10.81％,30～39岁组6例,占16.22％,40～49岁组9例,占24.32％,≥50岁有12例占32.43％.对HCV阳性者重叠其他型别肝炎病毒感染分析,单独HCV阳性2例,占5.41％,伴有HAV-IgG抗体阳性35例,占94.59％,伴有HEV-IgG抗体阳性10例,占27.03％,伴有HBsAg、HBcAb和HbeAb阳性者2例,占5.41％,该2例HAV和HEV抗体也阳性.结论 六省市(区)人群HCV感染率均在1％以下,感染者年龄以50岁以上最高,决大多数HCV阳性者重叠有其他型别的肝炎病毒感染.     

Prevalence of active infection by herpes simplex virus type 2 in patients with high-risk human papillomavirus infection: A cross-sectional study

The aim is to determine the prevalence of active infection by herpes simplex virus type 2 (HSV-2) among Mexican women with high-risk human papillomavirus (HR-HPV) cervical infection, recruited from public gynecology and colposcopy services. In a cross-sectional study, HSV-2 antibodies, HSV-2 DNA, and HR-HPV DNA were quantified. Significant differences in HSV-2 seroprevalence and HSV-2 active infection rates were found between negative and positive HR-HPV cases. HSV-2 seroprevalence was 28.15% and 16.1% (P = .0001), while HSV-2 active infection rates were 6.83% and 0.62% (P = .001) for positive and negative HR-HPV groups, respectively. The risk of HSV-2 seropositivity was 1.7 times greater for HR-HPV-positive cases (P = .02). Similarly, HR-HPV-positive cases were nine times more likely to have an HSV-2 active infection than HR-HPV-negative cases (P = .03). High HSV-2/h-HPV coinfection rates were observed among women recruited from public gynecology and colposcopy services. The main factors related to an HSV-2 active infection are a history of risky sexual behavior and HR-HPV infection. The prevalence of HSV-2 active infection among positive HR-HPV subjects indicate that these infections constitute an important group of STIs in Mexico.     

单克隆抗体作单纯疱疹病毒糖蛋白的特性鉴定以及临床应用

应用本室制备的8株抗单纯疱疹病毒糖蛋白单克隆抗体(抗HSV McAb)进行了系列研究,①鉴定出一种新的HSV糖蛋白g30K、gC的一种新形式,以及gC和gD;②建立了HSV感染的临床标本作抗原抗体分型检测的McAb-ELISA双夹心法,检测961份标本效果良好,并己在全国各地一些医疗单位推广使用;③证明McAb治疗家兔实验性单纯疱疹性角膜炎(HSK)有显著效果,并探讨其治疗机理主要为中和作用和ADCC效应;④对部分志愿者用McAb治疗单纯疱疹性角膜炎、妇女生殖器疱疹和小儿口腔疱疹性糜烂,取得明显疗效。     

Performance of a novel test for IgM and IgG antibodies in subjects with culture-documented genital herpes simplex virus-1 or -2 infection

Novel tests (BioPlex) for herpes simplex virus-1 (HSV-1) and HSV-2 IgG were compared with HerpeSelect HSV-1 and HSV-2 ELISAs for type-specific IgG. The sensitivity and specificity of BioPlex HSV-1 IgG were 94% (84/89) and 96% (119/124), respectively, with unselected sera, while the sensitivity and specificity of BioPlex HSV-2 IgG were 92% (109/118) and 98% (95/97), respectively. BioPlex IgM was compared with Diamedix IgM against sera from patients with culture-documented genital herpes. The test results were concordant in 81% of sera from HSV-1 patients and in 90% of sera from HSV-2 patients. Use of BioPlex IgM in addition to BioPlex IgG tests increased HSV-2 seroconversion detection from 47% of subjects to 70%. Use of Diamedix IgM in addition to Focus IgG ELISA increased HSV-2 detection from 40% of subjects to 70%. IgM was detected by BioPlex in 63% of sera from patients with early HSV-2 infection (< 30 days) and in 59% of sera by Diamedix. IgM was also detected in a large proportion of sera from subjects with established HSV-2 infection (33% by BioPlex and 29% by Diamedix). Addition of IgM testing substantially increased the ability to detect seroconversion early in infection. IgM is an indicator of recent infection only in subjects who lack detectable IgG.     

Performance of HerpeSelect and Kalon assays in detection of antibodies to herpes simplex virus type 2

The performances of commercial enzyme-linked immunosorbent assays (ELISAs) in detecting herpes simplex virus type 2 (HSV-2) antibodies have been inconsistent for African and human immunodeficiency virus (HIV)-positive populations. We compared the performances of the HerpeSelect and Kalon glycoprotein G2 ELISAs for patients with genital ulcer disease in Ghana and the Central African Republic. Sera from 434 women were tested with the HerpeSelect assay, and a subsample (n = 199) was tested by the Kalon assay. Ulcer swabs and cervicovaginal lavage samples were tested for HSV-2 DNA by PCR. HSV-2-seronegative women with detectable genital HSV-2 DNA were retested for HSV-2 antibodies 14 and 28 days later by the two ELISAs. A total of 346 (80%) women were positive by HerpeSelect at baseline, and 225 (54%) had detectable genital (lesional or cervicovaginal) HSV-2 DNA. Sixty-six (19%) HerpeSelect-positive samples had low-positive index values (1.1 to 3.5), and 58% of these samples had detectable genital HSV-2 DNA. Global agreement between the two serological assays was 86%. Concordance was high (99%) for sera that were negative by HerpeSelect or had high index values (>3.5). Defining infection detected by HSV-2 DNA PCR and/or Kalon assay as true infection, 71% of sera with low-positive index values were associated with true HSV-2 infection. Twenty-five women were identified as having nonprimary first-episode genital HSV-2 infection. Rates of HSV-2 seroconversion at day 14 were 77% (10/13 patients) by HerpeSelect assay and 23% (3/13 patients) by Kalon assay, with four additional seroconversions detected by Kalon assay at day 28. HIV serostatus did not influence assay performance. Low index values obtained with the HerpeSelect assay may correspond to true HSV-2 infection, in particular to nonprimary first episodes of genital HSV-2 infection, and need to be interpreted in the context of clinical history.     

Secreted portion of glycoprotein g of herpes simplex virus type 2 is a novel antigen for type-discriminating serology

The secreted portion of glycoprotein G (sgG-2) of herpes simplex virus type 2 (HSV-2) was evaluated as a novel antigen in an enzyme-linked immunosorbent assay (ELISA) format for detection of type-specific immunoglobulin G (IgG) antibodies in HSV-2-infected patients. The results were compared with those obtained by a commercially available assay, the HerpeSelect 2 ELISA (the FOCUS2 assay). Five different panels of sera were analyzed: panel A consisted of 109 serum samples from patients with a culture-proven HSV-1 infection that were Western blotting (WB) negative for HSV-2; panel B consisted of 106 serum samples from patients with a culture-proven recurrent HSV-2 infection that were WB positive for HSV-2; panel C consisted of 100 serum samples with no detectable IgG antibodies against HSV-1 and HSV-2; panel D consisted of 70 HSV-2 negative "tricky" serum samples containing antinuclear IgG antibodies or IgM antibodies against other viruses or bacteria; and panel E consisted of consecutive serum samples from 21 patients presenting with a first episode of HSV-2-induced lesions. When sera in panels A to C were analyzed, the sgG-2 ELISA and the FOCUS2 assay both showed sensitivities and specificities of >or=98%. In total, among the samples in panel D, 13 serum samples (19%) were false positive by the FOCUS2 assay and 1 serum sample (1.4%) was false positive by the sgG-2 ELISA. When the sera in panel E were analyzed, the sgG-2 ELISA detected seroconversion somewhat later than WB or the FOCUS2 assay did. We conclude that sgG-2 induces an HSV-2 type-specific antibody response and can be used for type-discriminating serology.     

High seroprevalence of herpes simplex virus type 2 infection in French human immunodeficiency virus type 1-infected outpatients

Using commercially available herpes simplex virus (HSV) type-specific serological diagnostic tests, HSV type 2 (HSV-2) antibody prevalence was assessed in two parallel prospective studies including 534 human immunodeficiency virus type 1 (HIV-1)-infected outpatients living in two areas of northern France. In the first cohort of 434 subjects, 223 (51%) individuals demonstrated a positive HSV-2 serological status while 66 (66%) of 100 subjects in the second cohort were seropositive for HSV-2 (51 versus 66%; P = 0.08). Among the 223 HSV-2-seropositive subjects identified in the first study cohort, only 22 (10%) had suffered from recurrent anogenital lesions during the past 12 months while 154 (69%) had no clinical history of herpesvirus infection. Our findings demonstrate high proportions of subclinical and undiagnosed HSV-2 infection in HIV-1-infected individuals and suggest that HSV type-specific serological testing in the French HIV-1-infected subpopulation could be an efficient strategy to diagnose clinically asymptomatic HSV-2 infections.     

The impact of incident and prevalent herpes simplex virus-2 infection on the incidence of HIV-1 infection among commercial sex workers in South Africa

This study investigated the impact of prevalent and incident HSV-2 infection on the incidence of HIV-1 infection in a cohort of female commercial sex workers in KwaZulu-Natal, South Africa. Prior to a vaginal microbicide trial, 416 women were screened for antibodies to HIV-1 and herpes simplex virus-2 (HSV-2) infections and a questionnaire was used to establish behavioral, social, and demographic characteristics. A total of 187 HIV-1-seronegative women were followed up at monthly intervals when blood was drawn and used to detect HIV-1 and HSV-2 antibodies. The median duration of follow-up was 2.2 years. At screening 50% of the women were HIV-1 seropositive and 84% were HSV-2 seropositive. The hazards of HIV-1 among women who were HSV-2 seropositive or seronegative throughout, or among those who seroconverted during the study, were not significantly different. When HSV-2 seroconversion was analyzed as a time-dependent covariate, the hazard ratio for HIV-1 seroconversion was 6.0 (95% CI: 2.6-14.0) times greater among women with incident than among women with prevalent HSV-2 infections. Drawing on other recent studies these data suggest that incident HSV-2 infection increases the risk of HIV-1 infection; the effect wanes with time since infection; and the effect is significantly greater for men than it is for women.     

Herpes simplex virus-1 and varicella virus infections in familial dysautonomia patients

Familial dystautonomia (FD) patients are deficient in type C fibers, suggesting that there may be a different pattern of infection and clinical presentation when infected by Herpes simplex virus type 1 (HSV-1) or Varicella-Zoster virus (VZV). These viruses infect and are reactivated in the periphery of the body through type C sensory nerve fibers. HSV-1 infects epithelial cells, penetrates into type C fibers, and migrates to the ganglia to generate latent infection. In reactivation, the viral DNA migrates through type C fibers, infecting the epidermis at the entry site. VZV infects through the respiratory tract, causing systemic viral infection and latency in the ganglia, from which it is reactivated and reaches the skin. The study was carried by clinical questionnaire and by HSV and VZV IgG antibodies on fifty-one FD patients and eighty matched controls. The questionnaire revealed that no FD patient had a history of clinical HSV-1 infection, compared to 15% in the control group (P < 0.05), while 50% FD patients had been infected by varicella, compared to 66% in the VZV control group. However in FD, VZV clinical manifestations were mild in comparison to controls. There was no difference in infection rates for some other viral diseases. HSV-1 antibodies were detected in 24% of the FD patients, compared to 38% in the control group (P < 0.1). VZV antibodies were similar in FD and controls (66%, 63%). We concluded that the rate of HSV infection in FD is low and clinical reactivation is rare. The rate of varicella infection appears to be the same for patients and controls, but in FD the clinical presentation is mild. We suggest that these differences are due to the lack of type C fibers in FD patients. J. Med. Virol. 54:158–161, 1998. © 1998 Wiley-Liss, Inc.     

Seroprevalence and determinants of herpes simplex type 2 infection in an STD clinic in Milan,Italy

A number of studies have shown that the seroprevalence of herpes simplex virus type 2 (HSV-2) is higher among persons attending clinics for sexually transmitted diseases (STD) than among the general population. The HSV-2 seroprevalence among STD patients, however, varies greatly among studies, possibly reflecting differences in the baseline prevalence of the infection among different general populations or in the distribution of risk factors. A cross-sectional study was carried out to determine the seroprevalence of and the risk factors for HSV-2 infection among 776 HIV-negative persons attending an STD clinic in Milan, Italy. All samples were tested with a commercial HSV type-2 specific gG ELISA test. The HSV-2 seroprevalence was 29.5% (95% CI: 26.3-32.7%). The seroprevalence increased with age, yet it did not differ by gender. Among persons with a current STD, the seroprevalence was 44.3%. At the multivariate analysis, older age was independently associated with HSV-2 infection. A self-reported history of genital herpes was predictive of HSV-2 infection. The agreement between history of genital herpes and HSV-2 seroprevalence was poor, however, stressing that in clinical practice, caution should be used in interpreting the presence or absence of a history of genital herpes as an indicator of the presence or absence of HSV-2 infection. Our data show that HSV-2 seroprevalence among persons attending an STD clinic in Italy is high; thus serological screening for HSV-2 might be advisable for STD patients.     

Diagnosing herpesvirus infections by real-time amplification and rapid culture

Procedures using real-time technique were developed to demonstrate the presence of herpes simplex virus type 1 (HSV-1) and HSV-2, varicella zoster virus (VZV), and cytomegalovirus (CMV) in miscellaneous clinical specimens. The assays were compared to rapid culture using centrifugation followed by detection with monoclonal antibodies. A total of 711 consecutive samples were collected from different patient groups. Throat swabs were obtained from transplant patients; dermal or oral specimens were collected from patients suspected for VZV or HSV infection. Genital specimens were taken from patients who attended the Clinic for Sexually Transmitted Diseases at the Dijkzigt Hospital Rotterdam presenting with symptoms of a primary genital ulcer. Nucleic acid extraction was carried out using a MagnaPure LC instrument. The amplification steps were performed on the ABI Prism 7700 sequence detection system. To monitor the process of extraction and amplification, a universal control consisting of seal herpesvirus type 1 (PhHV-1) was added to the clinical specimens. By culture 127 of 668 (19%) samples were positive for HSV-1, 72 of 668 (10.8%) specimens were positive for HSV-2, and 17 of 366 (4.6%) were positive for VZV. Using real-time amplification the numbers of positive specimens were 143 of 668 (21.4%), 97 of 668 (14.5%), and 27 of 366 (7.4%), respectively. Eighty-six specimens were tested for CMV, 12 (14.0%) were positive by culture, and 17 (19.8%) were positive by real-time PCR. The clinical data of the patients with discrepant results were reviewed thoroughly. In all cases the patients with only real-time PCR-positive results could be considered as truly infected. We concluded that the real-time amplification technique is suitable for the detection of human herpesvirus infection. It offers a semiquantitative and reliable assay with a quick result that is more sensitive than rapid culture, especially for the diagnosis of HSV-2 and VZV infections.