梅毒血清试验进展与临床关联

阐述螺旋体抗原与非螺旋体抗原研究进展及现今国内梅毒血清试验常用方法。各种试验的阳性或阴性结果与临床有密切关联。生物性假阳性发生的疾病及排除的方法。不同种类的抗梅药物致使梅毒血清反应阴转率也有差别。     

血清标本10546份非梅毒螺旋体抗原血清学试验假阳性结果分析

目的了解非梅毒螺旋体抗原血清学试验假阳性发生率及原因。方法对本院2007-2008年门诊及住院病人送检的血清标本进行快速血浆反应素(RPR)环状卡片试验和梅毒螺旋体颗粒凝集试验(TP-PA),对结果进行统计学分析。结果 2007-2008年合计检测了10546份血清标本,RPR假阳性率为0.21%(15/7002)。2007年(0.32%)与2008年(0.11%)的RPR假阳性率差异无统计学意义(P0.05)。15例RPR假阳性血清标本中10例为有高危性行为的常规检查者,2例为孕妇,皮肌炎、尖锐湿疣和龟头破溃者各1例。结论 RPR假阳性有一定的发生率。建议RPR阳性者以TPPA等梅毒螺旋体抗原血清学试验进行确认。     

神经梅毒

各期梅毒均可发生中枢神经系统损害.实验室检查显示非梅毒螺旋体抗原试验及梅毒螺旋体抗原试验阳性,脑脊液性病研究实验室玻片试验(VDRL)阳性,脑脊液白细胞计数≥10x106/L、蛋白定量>500 mg/L,可诊断为神经梅毒.水剂青霉素是目前治疗神经梅毒的最好选择.脑脊液中细胞计数是疗效观察的敏感指标,并发HIV感染者脑脊液异常时间可能延长.     

几种主要STD实验诊断方法的应用评价

在评价某试验方法的准确性时,常用敏感性和特异性这两个指标。若某法敏感性高而特异性低,则假阳性率高;敏感性低而特异性高则假阴性率高。一、梅毒本病由梅毒螺旋体引起,表现为皮肤粘膜及内脏的病变。 (一)使用非螺旋体抗原的非特异试验这些试验用从牛心肌中提取的脂质作为抗原,检测血清中的心脂质抗体。原理是由     

间质型神经梅毒6例临床分析

目的：总结间质型（脑膜及血管型）神经梅毒的临床特征，以提高对该病的认识。方法：回顾性分析6例间质型神经梅毒的临床表现、实验室检查和影像学特点。结果：6例间质型神经梅毒主要以脑卒中起病，实验室检查脑脊液示3例快速血浆反应素试验阳性，5例梅毒螺旋体抗原凝集试验阳性，6例脑脊液蛋白异常；头颅MRI表现脑动脉炎或脑梗塞等改变。结论：间质型神经梅毒临床表现多急性发病，影像学检查缺乏特异性，脑脊液梅毒螺旋体抗原凝集试验可能可以在神经梅毒中的应用，神经梅毒的诊断应结合临床表现及辅助实验检查综合分析。     

ELISA抗梅毒螺旋体抗原的特异性IgG/IgM抗体在诊断梅毒中的应用

目的探讨酶联免疫吸附测定(ELISA)抗梅毒螺旋体抗原的特异性IgG/IgM抗体在诊断梅毒中的应用。方法临床诊断或怀疑为梅毒的97例患者和74例非梅毒患者的血清同时接受ELISA抗梅毒螺旋体抗原的特异性IgG/IgM抗体和TPPA试验两种检测。结果ELISA抗梅毒螺旋体抗原的特异性IgG/IgM抗体具有较高的敏感性和特异性。结论ELISA抗梅毒螺旋体抗原的特异性IgG/IgM抗体适合于大样本的筛查和确诊。     

一种梅毒螺旋体重组抗原胶体金的制备

目的制备一种可以特异、准确的检测血清中梅毒螺旋体抗体的胶体金免疫层析试纸。方法利用原核表达系统构建一种包含梅毒螺旋体多个免疫表位的重组抗原B1,采用ELISA的方法对重组抗原进行活性检测。胶体金标记重组抗原B1,制备胶体金免疫层析试纸。结果重组抗原B1在BL21表达载体中有较高表达,ELISA结果显示B1与梅毒患者血清能特异、敏感的结合,与乙型肝炎病毒血清及阴性血清无交叉反应。应用B1蛋白制备的胶体金诊断试纸检测100份梅毒标准血清,结果均为阳性,与乙型肝炎患者血清及健康人群血清无交叉反应。结论梅毒螺旋体胶体金快速诊断试纸具有良好特异性和敏感性,可以快速、准确的诊断梅毒螺旋体的感染。     

6例麻痹性痴呆患者脑脊液检测结果分析

回顾性分析6例麻痹性痴呆患者脑脊液的特点并复习相关文献.结果:6例均为中年男性,脑脊液快速血清反应素试验(RPR)、梅毒螺旋体抗原凝集试验(TPPA)阳性,脑脊液主要变化为细胞、蛋白升高,免疫球蛋白升高,葡萄糖和氯化物基本正常.对血清RPR、TPPA 阳性的中年渐进性痴呆患者,脑脊液检查有助于神经梅毒的诊断,对其早期诊治有重要的临床参考价值.     

评价螺旋体乳胶快速反应实验检测梅毒血清的特异性和敏感性

梅毒的诊断依靠临床及实验室检查。目前实验室常规进行快速血浆反应素环状卡片试验(RPR),是一种筛查试验;用死的或活的梅毒螺旋体来检测抗梅毒螺旋体抗体,包括荧光密螺旋体抗体吸收试验(FTA-ABS)和梅毒螺旋体血球凝集试验(TPHA),这两者是确诊试验。除极早期梅毒外,各期梅毒均能得到诊断。心拟脂试验(如RPR)可检测到感染3～5周后的抗体,敏感性高,但缺乏特异性,可导致假阳性结果(如自身免疫性疾病等)。螺旋体试验(FTA-ABS,TPHA)敏感     

一期梅毒患者梅毒螺旋体明胶颗粒凝集试验的检测

梅毒螺旋体明胶颗粒凝集试验(TPPA)、快速血浆环状卡片反应素试验(RPR)和梅毒螺旋体暗视野检查(TP)等已作为常规试验检查梅毒患者。由于梅毒患者感染时间和就诊时间的差别及其他因素影响试验,使结果的敏感性不同。我们根据临床诊断,对TPPA、RPR和TP3项试验检测一期梅毒硬下疳患者血清和梅毒螺旋体的敏感性进行了分析。     

Antiphospholipid antibodies: new data in the pathogenesis of collagenosis

Antiphospholipid antibodies (APAB) were first detected as a result of the disturbances they cause in routine biological tests. These effects include lupus anticoagulant (LA) in APTT test and false serological reactions for syphilis in VDRL test. With more sensitive techniques using purified phospholipid antibodies (ELISA, RIA) antibodies directed specifically against cardiolipidin or other phospholipids can be assayed. Beside being responsible for positivity of the VDRL test in the context of syphilis, APAB (false serological reactions for syphilis, LA, anticardiolipin antibodies) have also been detected in systemic lupus erythematosus (SLE) and lupus-like syndromes, after intake of certain drugs, and, more rarely, in a number of diseases (table I). The specificity of APAB detected in syphilis (antiphosphatidylcholine) is different from that of APAB detected in SLE (antiphosphatidylserine and anticardiolipin). A given APAB can react with several phospholipids, but such crossreactions do not occur haphazardly with all phospholipids. Crossreactions are observed between APAB and polynucleotides on the one hand, and between antinuclear antibodies and phospholipids on the other. This pattern has led to speculate that APAB are directly involved in the pathogenesis of SLE; however since these crossreactions are not extensive, such speculation is controversial. The presence of APAB in the form of LA, false serological reactions for syphilis or anticardiolipin antibodies is associated with a number of problems including recurrent thrombosis, repeated abortions, thrombocytopenia and positive Coombs' tests or, more rarely, pulmonary hypertension, migraine, epilepsy, chorea, and transverse myelitis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reactivity of lymphocytes from patients with syphilis towards T. pallidum antigen in the leucocyte migration and lymphocyte transformation tests.

The reactivity of lymphocytes to Treponema pallidum antigen was studied before and after treatment in nine patients with early syphilis using a leucocyte migration test and a lymphocyte transformation test. Lymphocyte reactivity was also investigated in six patients treated for syphilis within the last 4 years, and in five untreated patients with a positive result to the T. pallidum immobilization test, but negative results to other serum tests for syphilis antibodies and without any known exposure to risk of infection by syphilis. Ten seronegative patients with different dermatological disorders served as a control group. A significant increase in lymphocyte reactivity to T. pallidum antigen was recorded in both tests in vitro after treatment. There was no difference in lymphocyte reactivity to T. pallidum antigen between the other patients studied and the control group. In early syphilis the spontaneous migration was found to be inhibited before treatment. Tuberculin skin tests were also performed and found to be suppressed in patients with primary and secondary syphilis. No difference in phytohaemagglutinin response was found between any of the groups. Plasma from patients with primary and secondary syphilis was found to change the in vitro reactivity of normal lymphocytes when stimulated with different mitogens.     

False positive reactions in confirmatory tests for syphilis in presence of antiphospholipid antibodies: misdiagnosis with prognostic and social consequences

False-positive reactions in the Venereal Disease Research Laboratory test are well known, whereas a positive fluorescent treponemal antibody absorption assay is rarely thought to be a false positive. The non-recognition of serological false-positive tests for syphilis may have negative prognostic and social implications.     

疑似梅毒患者血液47KDa和15KDa膜免疫原基因的测定

目的研究巢式聚合酶链反应(PCR)在排除梅毒血清学试验假阳性中的作用。方法用巢式PCR和常规PCR扩增90例疑似梅毒患者全血中梅毒螺旋体(TP)的47KDa和15KDa的膜免疫原基因,并与血清学方法(RPR,TPPA)的检验结果作比较。结果巢式PCR检测TP的敏感性和特异性分别为93.33%和96.67%,两者符合率为96.67%;两种PCR方法检测结果差异有统计学意义(P<0.01),巢式PCR敏感性高于常规PCR。结论巢式PCR检测全血中TP较常规PCR灵敏,通过检测47KDa和15KDa的膜免疫原基因,有助于排除梅毒血清学试验假阳性,提高了梅毒诊断的准确率。     

Specificity of the Treponema pallidum haemagglutination test. Analysis of results.

The automated haemagglutination assay using Treponema pallidum antigen (AMHA-TP) and the Venereal Disease Research Laboratory (VDRL) test were used to examine 330 163 sera. Reactive results were checked by the fluorescent treponemal antibody-absorption (FTA-ABS) test. When isolated reactivity or non-reactivity in the AMHA-TP test was investigated an estimated margin of error of 0.7% probably wrongly non-reactive and 0.008% presumably false non-reactive results were found. These figures were confirmed by randomised FTA-ABS tests on 504 sera with repeat AMHA-TP tests. The latter is therefore still the most reliable and practicable method for mass screening for syphilis.     

3种梅毒非特异性试剂检测脑脊液的结果分析

目的通过用3种试剂对患者脑脊液进行梅毒非特异性试验结果的分析,优选出神经梅毒的诊断试剂。方法使用VDRL1,VDRL2和RPR试剂同时对140例梅毒患者脑脊液标本进行梅毒非特异性试验。结果 140例脑脊液标本,检出TPPA阳性且VDRL或RPR定性阳性的合计26例,VDRL2和VDRL1,VDRL2和RPR定性实验结果比较,差异均有统计学意义(P均0.05)。结论用脑脊液检查进行神经梅毒的诊断和鉴别诊断时,RPR不能代替VDRL,且VDRL2优于VDRL1,建议使用VDRL试剂2。     

Syphilis serology in patients with primary syphilis and non-treponemal sexually transmitted diseases in southern Africa.

The reactivity of a non-specific reagin (RPR) test and a specific treponemal (FTA-ABS) test were determined in 21 patients with primary syphilis, 430 patients with proven non-treponemal genital ulcerations and 719 patients with acute urethritis presenting at a clinic for sexually transmitted diseases in southern Africa. Excluding those 21 cases of primary syphilis, 358 of 1149 tests performed (31%) were found to be reactive by at least one test. The rate of false positive RPR tests was very low (0.02%). Significantly higher rates of seropositivity were detected in patients with genital ulcerations than in patients with acute urethritis. The highest rates were detected among patients with proven lymphogranuloma venereum (34% RPR positive, FTA-ABS positive; 19% RPR negative, FTA-ABS positive). The geometric mean titres (GMT) of positive RPR tests in non-treponemal infections were found to be lower than in darkfield positive cases of genital ulcer disease.     

IgM rheumatoid factor removal and performance of the FTA-ABS (IgM) test in congenital syphilis.

OBJECTIVE--To determine the performance of the FTA-ABS (IgM) test in congenital syphilis after eliminating interference by IgM rheumatoid factor (RF) and preventing competitive inhibition by IgG. DESIGN--The FTA-ABS (IgM) test was carried out before and after RF removal (achieved by immunoprecipitation of the IgG) in infants with congenital syphilis and controls. SETTING--Newborns delivered in the Peninsula Maternal and Neonatal Services in Cape Town and infants presenting at Red Cross War Memorial Children's Hospital. SUBJECTS--Infants with congenital syphilis aged 0-4 months were divided into those with clinical signs at presentation and those who were asymptomatic at delivery. In addition, patients without congenital syphilis but with similar clinical signs at presentation were investigated as were control infants. OUTCOME MEASURE--The diagnosis of congenital syphilis was based on the criteria suggested by Kaufman et al (1977). RESULTS--Amongst symptomatic infants with congenital syphilis the FTA-ABS (IgM) test was positive in 34 (92%) of 37 cases prior to abolishing the RF effect and in 29 (78.4%) of 37 cases afterwards (p = 0.19). In 12 cases of congenital syphilis who were asymptomatic at birth, 10 had positive FTA-ABS (IgM) tests before RF removal and only three had positive tests afterwards (p = 0.006). False positive tests were not found amongst 15 symptomatic infants whose clinical features mimicked those of the infants with congenital syphilis. Among 51 healthy infants the test had a false-positive rate of 2% in newborns and 13% in older infants. The false positive reactions were eradicated by IgG precipitation. CONCLUSIONS--Following IgG and RF removal there was an improvement in the specificity of the FTA-ABS (IgM) test but this was at the expense of a loss of sensitivity, particularly in asymptomatic newborns. For newborns, if the FTA-ABS (IgM) test was positive, the patient was likely to require treatment for congenital syphilis, regardless of whether the result was due to the presence of RF or specific IgM.     

Syphilis serology and HIV infection in Harare, Zimbabwe

OBJECTIVE: To determine the reliability of serological tests in detecting syphilis in a factory worker cohort and examine the impact of concurrent HIV infection on serological tests for syphilis. METHOD: Reactions to non-treponemal and treponemal antigens were tested using sera from a cohort of 3401 factory workers in Harare, Zimbabwe. The participants consented to regular testing for syphilis, by VDRL, and HIV using two ELISAs. All sera from men who were VDRL positive, and a random sample of VDRL negative sera, were tested by RPR, TPHA, and where appropriate FTA-Abs. From the results, men were defined as having no syphilis, active syphilis, incident syphilis, historic syphilis, or giving biological false positive reactions. RESULTS: 709 sera were examined from 580 men. There were 78 cases of active syphilis in the cohort, giving a prevalence of 2.3%, and the seroincidence was 0.25 per 100 person years of follow up. The prevalence of HIV in the cohort was 19.8%. There was a strong association between syphilis, whether active, incident or historic, and HIV seropositivity. With both HIV positive and negative sera the negative predictive values of VDRL and RPR were > 99.9% while the positive predictive value for VDRL (30%) was lower than for RPR (39%). Biological false positive reactions were detected in 0.5% of the cohort, with in most cases a transient rise in VDRL titres up to < 1/16. Higher false positive titres occurred in five men, each of whom was HIV positive. CONCLUSIONS: The VDRL is reliable in detecting possible cases of syphilis even in a community with a high prevalence of heterosexually transmitted HIV. There is need, however, for confirmatory tests. The prevalence of syphilis in this cohort is very low in comparison with other countries in southern Africa, but is consistent with recent data from Harare. Despite a strong association between syphilis and HIV it was clear that syphilis could not be counted as a major factor fueling the HIV epidemic in Zimbabwe.     

Serological tests for syphilis in Saudi Arabia.

A total of 6684 sera were initially screened for syphilis by the Venereal Disease Research Laboratory (VDRL) test and the Treponema pallidum haemagglutination assay (TPHA). Reactive sera from either or both these tests were tested for confirmation by the fluorescent treponemal antibody-absorbed (FTA-ABS) test. VDRL biological false positive reactors were detected in 0.5% of the total sera examined, with 0.4% and 0.8%, respectively, obtained in pregnant women and blood donors. Eight sera (0.1%) were found to be positive in the TPHA test alone. An overall positivity of 2.7% for syphilis was detected, with a 0.85% positivity in antenatal patients. Infection with T pallidum seemed to be more common in men than in women (1.6:1) and predominated in the age group 20-39 years. Serological testing of sera from 26 mother and infant pairs allowed one case of congenital syphilis to be detected by FTA-ABS (IgM) and identified VDRL biological false positivity in seven infants.