ELISPOT检测在结核性脑膜炎诊断中的研究

目的利用酶联免疫斑点法(ELISPOT)检测脑脊液中结核抗原特异性的INF-γ分泌细胞数量,研究其在结核性脑膜炎诊断中的价值。方法对104例结核性脑膜炎患者和40例非结核性脑膜炎患者进行脑脊液与血的ELISPOT检测,并与脑脊液抗酸染色涂片、结核菌培养、荧光定量PCR、结核抗体检测进行敏感度和特异性的比较。结果脑脊液ELISPOT检测的敏感度为67.3%、特异性为89.2%,PV+值94.3%优于其他检测方法,差别具有统计学意义(P0.05),脑脊液ELISPOT检测具有较好的诊断价值(Kappa=0.856,P0.001)。结论脑脊液ELISPOT检测对结核性脑膜炎诊断的敏感度及特异度高,有望成为快速诊断结核性脑膜炎的一种方法。     

性病门诊单纯疱疹病毒2型特异性抗体血清学调查

目的了解性病门诊患者单纯疱疹病毒2型(herpes simplex virus type 2,HSV-2)感染情况。方法用ELISA法检测我院性病门诊1215例患者HSV-2型特异性抗体,并对不同性别和不同年龄组的HSV-2 Ig g G抗体阳性情况进行对比研究。结果 1215例患者中,HSV-2 Ig g G抗体阳性者共291例,阳性率为23.95%,男性阳性率为21.34%(197/923),女性阳性率为32.19%(94/292),女性阳性率高于男性,二者比较差异有统计学意义(P0.05)。年龄以31~40岁最多,为119例(男性81例,女性38例);其次为21~30岁,为85例(男性54例,女性31例)。结论 HSV-2型特异性抗体检测率存在男女差异,女性高于男性。生殖器疱疹易发于21~40岁性活跃人群。HSV-2型特异性抗体血清学检测具有快速、简便、灵敏度高等优点,适用于临床检测。     

抗核小体抗体对儿童系统性红斑狼疮诊断价值及与疾病活动度的相关性研究

目的 评价抗核小体抗体(AnuA)单独及与抗核抗体、抗Sm抗体和抗双链DNA(dsDNA)抗体联合检测对儿童系统性红斑狼疮(JSLE)的诊断价值,探讨AnuA与临床特点的相关性.方法 选取80例JSLE患者,56例非SLE的儿童疾病对照组,包括结缔组织病、白塞病、脊柱关节病、类风湿关节炎、皮肌炎、硬皮病,并留血清待检.采用酶联免疫吸附法(ELISA)检测血清AnuA,间接免疫荧光法检测抗核抗体;联合ELISA法和间接免疫荧光法检测抗dsDNA抗体,其中任一种检测方法结果阳性即判为阳性;免疫印迹法检测抗Sm抗体,并记录JSLE患者的临床特点.收集同期临床资料及相关实验室检查结果,进行SLE疾病活动指数(SLEDAI)评分,分析JSLE患者AnuA抗体与上述指标的相关性.结果 AnuA对JSLE诊断的敏感度为76.25％,特异度为98.21％.AnuA与抗dsDNA抗体及抗Sm抗体联合检测的敏感度分别为83.05％、86.44％,特异度96.43％、98.21％,敏感度明显高于二者单项检测的敏感度,差异有统计学意义(P＜0.01).AnuA与红细胞、血红蛋白呈负相关(r值分别为-0.499、-0.503,P值分别为0.000、0.000),与红细胞沉降率(ESR)、血尿、低补体、抗dsDNA抗体呈正相关(r值分别为0.388、0.227、0.303、0.531,P值分别为0.000、0.042、0.006、0.000),与SLEDAI评分呈正相关(r=0.303,P=0.000).结论 AunA对JSLE的诊断具有较高的敏感度和特异度,与抗dsDNA抗体和抗Sm抗体联合检测可以提高诊断的敏感度.AunA不仅可以作为JSLE的诊断指标,还可以评估疾病活动性,具有较高的临床应用价值.     

性病门诊妇女宫颈糜烂与单纯疱疹病毒2型活动感染关系探讨及临床治疗初探

目的 探讨性病门诊妇女宫颈糜烂与单纯疱疹病毒2型(HSV-2)的活动感染的关系，并初步探讨用干扰素栓治疗的临床效果。方法 用酶联免疫吸附试验(ELISA)对110例不同程度(Ⅰ、Ⅱ、Ⅲ度)宫颈糜烂病人和29例正常对照检测血清HSV-2型特异性IgM抗体。同时用ELISA法对病例组和对照组宫颈糜烂面分泌物和宫颈表面分泌物检测HSV-2抗原。HSV-2抗原检测阳性的患者用干扰素栓治疗。结果 110例不同程度宫颈糜烂病人HSV-2型特异性IgM抗体阳性率为40．91％(45／110)，宫颈糜烂面分泌物HSV-2抗原阳性率为34．55％(38／110)，抗原抗体同时阳性的感染率为20．00％(22／110)，均显著或非常显著地高于对照组(P=0．012，0．0003，0．0076)。在病例组中，HSV-2抗原检出率随糜烂程度加重而增高。38例抗原阳性患者用干扰素栓治疗痊愈显效率为73．68％，病毒抗原转阴率为71．10％。结论 性病门诊妇女宫颈糜烂的产生与HSV-2的原发活动感染有关，用干扰素栓治疗有效。     

分泌蛋白MPT64鉴定结核分枝杆菌复合群的应用研究

v、牛分枝杆菌和非洲分枝杆菌为阳性,24株非MTB均为阴性;mpt64基因扩增的结果与ELISA法检测结果完全相符.ELISA法检测MTB临床分离株的敏感度为97.3%(110/113),57株非MTB均未检出MPT64蛋白,检测特异度为100.0%.基因扩增MTB中2株为阴性,非MTB均为阴性.结论 采用ELISA法检测分泌蛋白MPT64来鉴定MTB复合群,是一种准确、快速和简便的方法,值得临床推广应用.     

GAD65反应性T细胞检测在成人隐匿性自身免疫糖尿病筛查中的应用研究

目的探讨酶联免疫斑点试验(ELISPOT)检测谷氨酸脱羧酶反应性分泌IFN-γT细胞(GAD65-T)在成人隐匿性自身免疫糖尿病(LADA)筛查中的应用。方法选择2014年至2016年福建医科大学附属泉州第一医院收治的LADA患者28例,2型糖尿病(T2DM)40例,健康对照20例,放射配体法检测GAD65-Ab,连续密度梯度离心法分离人外周血单个核细胞(PBMC),ELISPOT法检测GAD65-T细胞。将40例T2DM分为GAD65-T细胞ELISPOT检测阳性及阴性组,比较两组的临床特征。结果 GAD65-T细胞数(中位数及95%CI)LADA组为7.2(1.9~11.1)个,T2DM组为3.6(2.9~6.0)个,对照组为0.9(0.2~1.7)个。LADA组的细胞数明显高于T2DM及对照组(P均0.01);T2DM较对照组具有更高频率的细胞数(P0.05)。以大于95%CI(对照组)判为阳性,LADA,T2DM及对照组的阳性率分别为71.4%(20/28)、32.5%(13/40)、0。40例T2DM亚组分析显示GAD65-T细胞ELISPOT检测阳性组较阴性组的患者BMI更低、血糖控制更差、空腹C肽水平更低。结论部分GAD65-Ab阴性的T2DM患者存在GAD65-T细胞,联合GAD65-T细胞ELISPOT及GAD65-Ab检测可明显提高LADA的检出率。     

酶联免疫斑点法检测肺结核患者特异性结核抗原多肽T细胞的应答

目的 评价我国酶联免疫斑点法(ELISPOT)用于诊断肺结核患者的应用价值.方法 结核特异性ELISPOT法检测76例诊断明确的肺结核患者外周血单核细胞.同时选择30例无结核接触史、胸部X线片无异常且PPI)皮试阴性的成年人作为对照.数据采用卡方检验和非参数Mann-Whitney检验.结果 76例患者总体阳性率为71.0%,健康对照组阳性率为6.7%.按照患者抗结核疗程分为治疗1个月内、治疗6个月内、治疗6个月以卜3组,阳性率分别为87.5%、74.1%和52.0%(X2=35.63,P=0.000).该技术在初治结核患者中的灵敏度为87.5%,特异度为93.3%,阳性预测值为91.3%,阴性预测值为90.3%.结论 结核特异性ELISPOT技术在我国初治(抗结核化疗疗程1个月内)的结核病患者中具有较高诊断价值.     

抗神经节苷脂抗体与神经精神性狼疮相关性研究

目的 探讨抗神经节苷脂抗体在神经精神性狼疮(NPSLE)中的意义.方法 采用酶联免疫吸附试验(ELISA)法,检测68份血清(包括NPSLE患者33例.无神经精神症状的狼疮患者35例)和18例脑脊液(包括NPSLE患者10例,非NPSLE的狼疮患者8例)中的抗神经节苷脂抗体IsG和IgM.结果 血清中抗神经节苷脂抗体IgG、IgM在系统性红斑狼疮患者中的阳件率分别为21%、24%,在NPSLE患者中的阳性率均为18%,在无神经精神症状的狼疮患者中的阳性率分别为22%、29%,二者在抗体水平和阳性率上差异无统计学意义(P＞0.05).脑脊液中抗神经节苷脂抗体IgG、IgM在系统性红斑狼疮患者中的阳性率分别为10/18、9/18,在神经精神性狼疮患者中的阳性率均为7/10、8/10,在非NPSLE的狼疮患者中的阳性率分别为3/8、1/8,二者在抗体水平和阳性率上差异有统计学意义(P＜0.05).结论 脑脊液中AGA与NPSLE密切相关.     

乙型脑炎病毒抗体捕获ELISA检测方法的初步建立

目的 建立检测乙型脑炎病毒抗体的抗体捕获ELISA方法.方法 以乙型脑炎病毒E蛋白抗原表位E39特异的单克隆抗体包被ELISA板,吸附乙型脑炎减毒活疫苗株SA14-14-2病毒粒子,再以吸附的病毒粒子捕获乙型脑炎病毒抗体,建立抗体捕获ELISA方法:同时与以乙型脑炎病毒细胞培养上清包被的间接ELISA方法进行对比,并检测临床血清样品105份.结果 间接ELISA方法背景无法消除,1:10、1:100、1:1000各稀释度的阳性血清与阴性血清吸光度(A)值相近,A阳性血清/阴性血清分别为1.02、0.99、1.13,均<2.1.而抗体捕获ELISA方法1:10、1:100稀释度的A阳性血清/阴性血清分别为3.57、2.94,均>2.1;1:1000稀释度的A阳性血清/阴性血清为1.42,<2.1,表明血清稀释度为1:100时可以显著区别乙型脑炎病毒感染的阳性血清与阴性血清,明显消除间接ELISA方法中背景过高的问题.抗体捕获ELISA方法检测105份临床血清样品,A阳性血清/阴性血清为0.257～0.321(0.262±0.050),均<2.1,为乙型脑炎病毒抗体阴性血清.结论 初步建立了乙型脑炎病毒抗体捕获ELISA方法,该方法特异性较高,对于建立乙型脑炎病毒群的快速鉴别诊断方法具有重要意义.     

急性病毒性脑炎220例临床特征与病原学分析

目的探讨急性病毒性脑炎(AVE)患者的临床特点与病原学的关系。方法回顾性分析220例确诊为AVE住院患者[随访时间最短150 d,最长18月,失访13例(5.9%);≤14岁179例(81.36%),≥15岁41例(18.64%)]的临床资料。结果 220例患者血液和脑脊液(CSF)均检测出相关病毒特异性抗体IgM阳性。依次为单纯疱疹病毒-1(HSV-1)146例(66.36%),腮腺炎病毒(PXV)21例(9.55%),柯萨奇病毒-A/B(CSV-A/B)19例(8.64%),带状疱疹病毒(VZV)12例(5.45%),巨细胞病毒(CMV)10例(4.55%),乙型脑炎病毒(JEV)7例(3.18%),EB病毒5例(2.27%)。临床治疗总有效率为90.46%,病死率为9.55%。结论血清和CSF检测有助于明确AVE病原学,及时早期应用有效抗病毒药物,可明显改善AVE预后。     

Herpes simplex virus type-1 meningoencephalitis showing disseminated cortical lesions

We report a 41-year-old man with meningoencephalitis associated with herpes simplex virus type 1 (HSV-1). The patient developed fever, headache and dysuria followed by generalized convulsion and neck stiffness, and the CSF showed pleocytosis. The titers of enzyme-linked immunosorbent assay against HSV measured 6 days after onset showed a significant rise; IgG antibody 4.89 (<0.2) and IgM antibody 1.45 (<0.8) in CSF, IgG antibody 46.1 (<2.0) and IgM antibody 1.76 (<0.8) in the serum. The antibody index for IgG was 0.50, and that for IgM was 4.2. CFS neutralization test showed HSV-1 antibody of x16 and HSV-2 antibody of 相似文献   

Correlation of herpes simplex virus types 1 and 2 with clinical features of infection.

Strains (338) of herpes simplex virus (HSV) were isolated in Stockholm during 1965-1974. By immunoelectroosmophoresis it was possible to identify all strains as either HSV type 1 (HSV-1) or 2 (HSV-2). No strains of intermediate antigenic type or with untypable characteristics were found. The antigenic type of HSV was correlated with body site and clinical features of infection. A case of severe, recurrent, abdominal pain in association with HSV-2 infection is described. In one patient with acute aseptic meningitis, both coxsackievirus A9 and HSV-2 were isolated from the same specimen of cerebrospinal fluid. Serology suggested a primary infection with coxsackievirus A9 and a recurrent HSV-2 infection. HSV-1 was isolated from specimens of cerebrospinal fluid. Serology suggested a primary infection with coxsackievirus A9 and a recurrent HSV-2 infection. HSV-1 was isolated from specimens of cerebrospinal fluid from two of four adults with HSV encephalitis.     

A Probable Case of Herpes simplex Encephalitis despite Negative PCR Findings Findings

A 54-year-old woman was admitted to the hospital suffering from fever and personality changes. Laboratory examination of her cerebrospinal fluid (CSF) showed 270 mononuclear cells, 30 polynuclear cells and a clinically low number of erythrocytes/mm³. Empirical clinical findings from this case suggested treatment with acyclovir. Magnetic resonance imaging (MRI) showed bilateral temporal hyperintense signals in T2-weighted images. PCR with specific primer for herpes simplex virus type 1 (HSV-1) and HSV-2 were negative. There was no elevation of oligoclonal antibodies specific to HSV in CSF after 2 weeks. Although we did not prove the presence of the agent microbiologically at the clinical onset of the disease, the MRI and electroencephalogram (EEG) findings, erythrocytes in CSF and the dramatic response to acyclovir therapy are suggestive of a diagnosis of herpes simplex encephalitis (HSE). Received: July 7, 2000 · Revision accepted: October 11, 2001     

Rapid diagnosis of viral infections in the central nervous system

Rapid diagnosis of viral infections in the central nervous system has become increasingly important. Antiviral treatment, prevention of spread of disease and differentiation from infections caused by agents sensitive to antibiotics may be the important consequences of a virus specific diagnosis gained early in the disease. The diagnosis can be obtained by detection of virus or viral antigen in the human specimen: herpes simplex virus by electron microscopy, immunofluorescence or immunosorbent assays in brain biopsies; rabies virus by immunofluorescence in corneal cells or skin and mucous membranes. The presence of measles or influenza antigens in nasopharyngeal secretions, shown by immunofluorescence or enzyme immunoassays, may diagnose an encephalitis caused by either of these viruses. Where suitable material is not available the detection of virus-specific IgM in a single serum specimen may be used for diagnosis. Mumps specific IgM activity is detected by enzyme-linked immunosorbent assay (ELISA) or indirect immunofluorescence techniques; tick-borne encephalitis (TBE) specific IgM by immunosorbent assays or by reduction of hemagglutination-inhibition (HI) titer by 2-mercaptoethanol treatment of serum. Reports have been given on the detection of IgM activity by ELISA also in other arboviral infections such as Japanese and LaCrosse encephalitis. The demonstration of an intrathecal production of virus-specific immunoglobulins may reveal the type of virus causing the infection in the central nervous system.     

Evaluation of three commercially available Japanese encephalitis virus IgM enzyme-linked immunosorbent assays

We evaluated performance of three commercial Japanese encephalitis virus (JEV) IgM antibody capture enzyme-linked immunosorbent assay (MAC ELISA) kits with a panel of serological specimens collected during a surveillance project of acute encephalitis syndrome in India and acute meningitis and encephalitis syndrome in Bangladesh. The serum and cerebral spinal fluid specimens had been referred to the Centers for Disease Control and Prevention (CDC) for confirmatory testing. The CDC results and specimen classifications were considered the reference standard. All three commercial kits had high specificity (95-99.5%), but low sensitivities, ranging from 17-57%, with both serum and cerebrospinal fluid samples. Specific factors contributing to low sensitivity compared with the CDC ELISA could not be determined through further analysis of the limits and dilution end points of IgM detection.     

Herpesvirus infections of the central nervous system

In recent years, advances in the diagnosis and treatment of herpes simplex encephalitis (HSE) have been achieved due to the prevalence of antiviral drugs and the introduction of the polymerase chain reaction (PCR) to test the cerebrospinal fluid. The several clinical forms of herpes simplex virus type 1 (HSV-1) infections of the central nervous system (CNS), including acute disseminated encephalomyelitis and brainstem encephalitis, have been clarified. However, fatal, prolonged, or relapsed cases are still observed, and early detection and appropriate treatment is necessary to lead to a good prognosis for these intractable HSE cases. In adult HSV-2 infections, meningitis and myelitis associated with genital herpes are common. In the past, HSV-2 myelitis has been reported as a form of fatal necrotizing myelopathy; however, using PCR and magnetic resonance imaging studies, mild surviving cases are increasingly likely to be identified. Meanwhile, various CNS syndromes resulting from the herpes group viruses, including varicella-zoster virus and Epstein-Barr virus have also been reported. These herpesviruses have several characteristics in common, e.g., they exist in the latent state and they occur in both mucocutaneous and CNS infections. Adult HSV-1 and -2 infections of the CNS are discussed together with other herpes group virus infections of the CNS.     

Herpes simplex esophagitis in the immunocompetent host

We report here a case of herpes esophagitis with Mallory-Weiss syndrome in an immunocompetent host. A 26-year-old man was admitted to our hospital because of common cold symptoms and eruptions on the body. On day 2 after hospitalization, the patient showed high-grade fever, odynophagia and hematemesis. Upper gastrointestinal endoscopic examination showed multiple ulcerations throughout the mid- and distal esophagus. Bleeding from a Mallory-Weiss tear was also seen. Follow-up endoscopic examinations showed whitish exudates on day 5. Histological examination of biopsy specimens showed Cowdry type A intranuclear inclusion bodies in epithelial cells. Positive staining of a specific antibody against herpes simplex virus-1 (HSV-1) was seen in the nuclei of esophageal epithelial cells. Primary HSV-1 infection was suspected because ELISA titers of serum IgM antibody against HSV-1 were high and titers of serum IgG antibody against HSV-1 increased from an almost cut-off ratio. A diagnosis of herpes esophagitis in an immunocompetent host was made. Our case is the first report of herpes esophagitis with Mallory-Weiss syndrome in the immunocompetent host. It is important to remind herpes esophagitis in cases of severe odynophagia even in immunocompetent hosts.     

Prevalence of herpes simplex type 2 antibodies and a clinical history of herpes in three different populations in Campinas City, Brazil.

OBJECTIVES: To determine the seroprevalence of herpes simplex virus type 2 (HSV-2) antibodies and the relation between the history of clinical herpes and the presence of type-specific HSV-2 antibodies in three different populations from the city of Campinas City, Brazil. POPULATION AND METHODS: One hundred and one college students, 96 patients with sexually transmitted diseases (STD), and 102 women at delivery were interviewed and blood samples were collected. Total HSV (HSV-1 and HSV-2) antibodies were screened by enzyme-linked immunosorbent assay (ELISA) and type-specific HSV-2 antibodies were detected by Western blot assay. RESULTS: Herpes simplex virus antibodies were detected in 66.3% of the students, 97.1% of the women at delivery, and 99.0% of the STD patients. Type-specific HSV-2 antibodies were detected in 6.9% of the students, 22.6% of the women at delivery, and in 53.1% of the STD patients. History of genital herpes was reported by none of the students, by one of the women at delivery, and by 11 of 51 (21.6%) STD patients who were HSV-2 seropositive. Four of the 45 (8.9%) seronegative STD patients reported a history of genital herpes. CONCLUSION: The prevalence of HSV-2 infection in Campinas City can be significantly affected by the characteristics of the population studied, as was shown in previous studies. The sensitivity of the history of genital herpes was low in the present series, stressing that prophylactic measures for vertical and horizontal transmission of HSV-2 should not be based only on a positive history of genital ulcers.     

Prevalence of herpes simplex virus antibodies in childhood and adolescence: a cross-sectional study

The changing spectrum of herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) infections makes it important to define the seroepidemiology of HSV. The object of this study was to determine the prevalence of HSV-1 and HSV-2 immunoglobulin G antibodies in a young Swedish population by investigating 2106 serum samples from people aged 0-19 y. Sera were tested in HSV type-specific enzyme-linked immunosorbent assays using glycoprotein G-1 (gG-1) and glycoprotein G-2 (gG-2) as antigens. The overall seroprevalence was 31% (95% CI 29-33) for HSV-1 and 0.5% (95% CI 0.2-0.9) for HSV-2. The HSV-1 seroprevalence was higher with increasing age, and significantly higher in the age cohort 15-19 y compared with 1-4-y-olds (37% vs 24%). The HSV-1 infection seemed to be acquired early in life. In the age cohort 1-2 y, the prevalence was over 20%, presumably reflecting an established viral infection. In adolescence the HSV-1 seroprevalence may reflect both oral and sexual transmission. The seroprevalence in the oldest age cohort did not differ significantly from that seen in a Swedish study in which sera were sampled from young girls in the 1970s.     

Evaluation of a near-patient test and 2 enzyme-linked immunosorbent assay-based assays for detecting anti-herpes simplex virus type-2 antibodies.

Several type-specific serologic assays for herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), based on glycoprotein G1 (gG1) and gG2, have recently been developed. These include immunodot (POCkit HSV-2) and enzyme-linked immunosorbent assay (ELISA). The diagnostic value of POCkit HSV-2, a near-patient test, and of 2 immunoenzymatic, type-specific assays was evaluated on 122 patients attending an STD clinic. Western blot was used as the reference test. The sensitivity of POCkit HSV-2 was good but the specificity was poor, so that in a population with low seroprevalence, a positive result is likely to be a false positive. Analysis of 2 currently available HSV type-specific ELISAs yielded results suggesting that the sensitivity of these tests may also be suboptimal.