聚合酶链法检测莱姆病患者尿液中伯氏疏螺旋体DNA(附17例报道)

目的:评估聚合酶链反应(PCR)检测莱姆病患者尿液中伯氏疏螺旋体DNA的诊断价值。方法:2004至2006年来自新疆莱姆病自然疫源地临床疑似莱姆病患者17例(均有蜱暴露史和莱姆病临床证据者)和6例以前确诊并治疗后完全恢复的莱姆病患者,共23例为病例组;另选择25例非疑似莱姆病患者为对照组。应用PCR方法检测尿液中伯氏疏螺旋体DNA,采用间接免疫荧光(IFA)检测血清伯氏疏螺旋体抗体。结果:①17例莱姆病患者临床表现呈多系统损害。12/17例(70.59%)为神经型(周围神经病3例,其中1例合并下肢瘀积性皮炎;游走性红斑并发莱姆脑病2例;脑膜脑炎2例;脑炎2例;脑膜炎、脑干炎和脊髓炎各1例);流感样症状2例,其中伴皮疹1例;余心脏型、精神障碍和肝病型各1例。②PCR伯氏疏螺旋体DNA检测阳性17/17例(总阳性率100%)。4例血清IFA试验伯氏疏螺旋体抗体阳性。以前确诊并治疗者6例及对照组25例尿PCR伯氏疏螺旋体DNA和血清IFA试验伯氏疏螺旋体抗体均阴性。③10例完成抗生素治疗后3个月复查,9例PCR伯氏疏螺旋体DNA检测阴性(包括3例晚期莱姆病患者),1例2年复查时仍阳性;4例血清伯氏疏螺旋体抗体阳性者复查均阴性。④早期莱姆病多采用几周的抗生素治疗方案。结论:PCR检测尿液中伯氏疏螺旋体DNA是莱姆病一个有价值的诊断工具。     

莱姆病神经系统表现

目的报道2例莱姆病神经系统表现并进行相关文献复习。方法对可疑病例进行全面检查，排除其他可能的疾病，并进行莱姆病相关检查。结果发现2例神经系统莱姆病，例1表现为多颅神经炎、脊髓病变、多发性神经根神经炎，例2表现为多发性神经根神经炎及周围神经病变。常规脑脊液检查正常。血清及脑脊液抗伯氏疏螺旋体抗体滴度分别为1：64~1:264及1：128~1:256。结论对原因不明的多颅神经炎、脊髓病变、周围神经病变应进行莱姆病相关化验，以利早期诊断、治疗。     

2000至2004年新疆维吾尔自治区莱姆病的监测

目的:监测莱姆病(LD)在新疆地区的流行情况与特征,为LD的防治提供决策依据。方法:2000至2004年采用间接免疫荧光抗体法(IFA)、蛋白印迹法(WB),对业已证实的新疆自然疫源地区2167人份的血清标本进行抗LD螺旋体抗体(IgM、IgG)检测;采用IFA、WB、ELISA试验对来自21个县、市的门诊与住院患者,随机抽样1443人份血清标本进行抗LD螺旋体抗体(IgM、IgG)检测作对照;共计3610份血清标本。结果:自然疫源地2167人份血清中IFA:IgM阳性数546人、IgG阳性数223人、总阳性数769人,总阳性率为35.49%。其中471人份血清同时作WB法,其IgM、IgG阳性数均为28人,阳性率为13.49%,与对照组总阳性率20.99%比,差异有统计学意义(P<0.01)。结论:新疆自治区LD自然疫源地平均感染率明显高于国内(1.5%和8.06%)水平,各年度的抗体水平呈递增趋势,应该加强对新疆LD的监测与防治。     

T-SPOT.TB法检测脑脊液单个核细胞对结核性脑膜炎早期诊断价值的研究

研究背景结核性脑膜炎为严重危害人类健康的中枢神经系统感染性疾病,传统的脑脊液分析和放射学检查方法敏感性和特异性较低,延误治疗时机,因此临床急需敏感性更高的快速检查方法用于结核性脑膜炎的诊断。本文采用T细胞酶联免疫斑点试验(ELISPOT)检测结核性脑膜炎患者外周血和脑脊液单个核细胞,比较两种检查方法诊断结核性脑膜炎的敏感性和特异性,以为结核性脑膜炎的早期诊断提供有效检查方法。方法分别收集结核性脑膜炎(30例)和非结核性脑膜炎(30例)患者外周血和脑脊液,分离提取单个核细胞,经冻存和复苏后采用结核杆菌感染T细胞斑点试验(TSPOT.TB)检测外周血和脑脊液中干扰素γ分泌的T细胞数目,分别计算其诊断结核性脑膜炎的敏感性和特异性。结果脑脊液ELISPOT试验显示,结核性脑膜炎组阳性检出率为93.33%(28/30)、非结核性颅内感染组为3.33%(1/30);检测灵敏度93.33%、特异度96.67%,假阳性率3.33%、假阴性率6.67%,阳性似然比28.33、阴性似然比0.07。外周血ELISPOT试验显示,结核性脑膜炎组阳性检出率为76.67%(23/30)、非结核性颅内感染组为13.33%(4/30),检测灵敏度76.67%、特异度86.67%,假阳性率13.33%、假阴性率23.33%,阳性似然比5.79、阴性似然比0.15。外周血ELISPOT试验阳性检出率与脑脊液阳性检出率比较,差异无统计学意义(Fisher确切概率法:P=0.254)。结论采用ELISPOT试验检测外周血和脑脊液单个核细胞结核杆菌感染率有助于提高结核性脑膜炎的诊断率,而且脑脊液单个核细胞ELISPOT试验检测效率呈高于外周血检测效率之趋势。     

脑脊液细胞学检查和病毒抗体检测对病毒性脑膜脑炎的诊断价值

目的:探讨脑脊液细胞学检测和病毒抗体检测对病毒性脑膜脑炎的诊断价值。方法:选择84例颅内感染患者分为:病毒性脑膜脑炎组32例,结核性脑膜脑炎组32例,化脓性脑膜脑炎组20例。结核性脑膜脑炎组和化脓性脑膜脑炎组作为对照。每例患者各取2次脑脊液,共168份脑脊液。分别行细胞学检测和ELISA法检测病毒系列抗体IgG、IgM。结果:病毒性脑膜脑炎的脑脊液细胞学超早期有明显的嗜中性粒细胞反应,随后出现激活的淋巴细胞反应;随着患者病情好转,激活的淋巴细胞逐渐减少;转而以淋巴细胞、单核细胞为主。病毒抗体IgG检测:病毒性脑膜脑炎组阳性率为86.7%,明显高于结核性脑膜脑炎组(48.3%)和化脓性脑膜脑炎组(15%);抗体IgM检测:病毒性脑膜脑炎组与结核性脑膜脑炎组和化脓性脑膜脑炎组比差异无统计学意义。结论:脑脊液细胞学检查快速、简便,是中枢神经系统病毒感染的重要指标之一;病毒特异性IgG抗体诊断病毒性脑膜脑炎的敏感度为54.7%,特异度为91.3%,是确诊病毒性脑膜脑炎病原的方法之一。     

脑脊液腺苷脱氨酶与血清5’-核苷酸酶在结核性脑膜炎诊断中的价值

目的探讨脑脊液腺苷脱氨酶(CSF-ADA)与血清5’-核苷酸酶(5’-NT)在结核性脑膜炎诊断与鉴别诊断中的价值。方法回顾性分析40例结核性脑膜炎患者(病例组)与36例病毒性脑膜炎患者(对照组)的一般资料、脑脊液、血液学及影像学检查结果,重点探讨CSF-ADA与血清5’-NT在结核性脑膜炎中的诊断价值。结果病例组CSF-ADA和血清5’-NT水平显著高于对照组(P<0.001)。根据CSF-ADA与血清5’-NT的接收者操作特性曲线(ROC),CSF-ADA>4.40 U/L或血清5’-NT>4.85 U/L被认为是诊断的阳性结果,显示出高的敏感性与特异性,CSF-ADA的敏感度为92.5%,特异度为89%;血清5’-NT的敏感度为95%,特异度为70%。当联合CSF-ADA与血清5’-NT诊断结核性脑膜炎,即CSF-ADA>4.40 U/L或(和)血清5’-NT>4.85 U/L,其敏感性最高,其灵敏度为97.5%,特异度为73.5%,阳性似然比(PLR)为3.68,阴性似然比(NLR)为0.03。结论在结核性脑膜炎的诊断中,CSF-ADA与血清5’-NT可以起到快速、简单、相对准确的诊断作用。当CSF-ADA或(和)血清5’-NT明显升高时,可尽早怀疑结核性脑膜炎,同时结合临床表现、其他脑脊液、血液学及影像学结果综合诊断。     

老年期痴呆患者脑脊液微管相关蛋白tau的检测

目的探讨老年期痴呆(SD)患者脑脊液(CSF)中微管相关蛋白tau(以下简称tau)的检测方法.方法采用双抗体夹心酶联免疫吸附(ELISA)法检测26例SD患者和30例非神经系统疾病患者的CSF-tau.结果两者浓度分别为473±279、151±60 pg/ml,有极显著差异(P<0.001);本法灵敏度为53.8%,特异性为83.3%,检出限60 pg/ml,组内变异1.2%～5.9%,组间变异1.7%～6.0%.结论采用双抗体夹心ELISA法检测SD患者CSF-tau的浓度可为诊断SD提供有价值的参考依据.     

降钙素原联合乳酸检测在术后细菌性颅内感染诊断中的应用

目的 探讨降钙素原（PCT）联合脑脊液乳酸（LA）检测在颅脑术后急性细菌性颅内感染（ABII）诊断中的应用价值。方法 回顾性分析2018年6月至019年6月收治的颅脑手术后发热的267例病人的临床资料。术后3~7 d出现体温超过≥38 ℃，即刻抽取静脉血，检测血常规、LA和PCT；行腰椎穿刺术检查脑脊液常规、生化、细菌培养、LA和PCT。结果 267例中，发生ABII有51例（感染组），无ABII216例（非感染组）。与非感染组相比，感染组体温明显增高（P<0.05），血清和脑脊液PCT均明显增高（P<0.01），感染组脑脊液LA明显增高（P<0.01）。血清PCT≥0.05 ng/ml诊断ABII的灵敏度和特异度分别为92.16%、91.20%，脑脊液PCT≥0.05 ng/ml诊断ABII的灵敏度和特异度分别为98.04%、97.69%。血清LA≥2 mmol/L诊断ABII的灵敏度为82.35%，特异度为26.39%；脑脊液LA≥3 mmol/L诊断ABII的灵敏度为96.08%，特异度为63.89%。结论 血清和脑脊液PCT对ABII均有较高的诊断价值，而血清和脑脊液LA诊断ABII的特异度欠佳。     

脑卒中患者抗心磷脂抗体的检测及分析

目的:探讨抗心磷脂抗体(ACA)与脑卒中的关系,为临床对脑卒中的诊断和预防提供依据。方法:用酶联免疫吸附试验(ELISA)测定378例脑卒中患者血清中的抗心磷脂抗体水平。结果:①脑梗死患者ACA的阳性率(31·6%)明显高于脑出血患者的ACA阳性率(9·38%)(P<0·001);②年龄<30岁和年龄≥30且≤50岁的脑出血、脑梗死患者ACA阳性率均显著高于50岁以上的脑出血、脑梗死患者ACA阳性率。结论:①在脑卒中患者中,ACA对脑梗死的发生比对脑出血的发生更为密切;②ACA与青、中年的脑卒中患者关系密切,及早进行ACA检测将有助于脑卒中的预防和诊治。     

酶联免疫斑点法和IS6110聚合酶链反应在早期诊断结核性脑膜炎中的价值

目的 探索一种敏感而特异的早期诊断结核性脑膜炎(TBM)的方法.方法 对25例TBM患者及作为对照的27例其他中枢神经系统感染性疾病患者、22例非感染性其他神经系统疾病患者同时应用酶联免疫斑点法(ELISPOT)检测脑脊液和外周血中的抗卡介苗(BCG)抗体分泌细胞数,应用PCR检测脑脊液中结核杆菌DNA的特异性重复插入片段IS6110,用酶联免疫吸附法(EUSA)检测脑脊液和外周血抗BCG抗体.结果 ELISPOT的敏感性(84.0%)明显高于IS6110PCR(75.0%)及ELISA(52.3%),三者的特异性分别为91.8%、93.7%和91.7%.TBM患者脑脊液抗BCG-IgG分泌细胞的个数在疾病早期最高,随病程的延长逐渐下降(t=-3.183,P=0.008).结论 ELISPOT对早期诊断TBM有一定的价值.     

Comparative study of four different assays for the detection of binding antibodies against interferon-beta

BACKGROUND: Binding antibodies (BAB) against interferon-beta (IFNbeta) are often determined as screening assays before performing an expensive and elaborate neutralizing antibody (NAB) test. METHODS: In this study, we compared four BAB tests, a western blot (WB), a direct binding enzyme-linked immunosorbent assay (ELISA) (dELISA), a capture ELISA (cELISA), and a commercial enzyme immuno-assay (EIA) in 325 multiple sclerosis patients with and without neutralizing antibodies to evaluate the sensitivity and specificity to detect NAB by receiver operating characteristics analysis. RESULTS: The area under the curve (AUC) values were 0.907 for the dELISA, 0.925 for the cELISA, and 0.776 for the EIA (P < 0.0001 for all). At a sensitivity of 95%, the specificity was approximately 30% in the dELISA, 55% in the cELISA, and 13% in the EIA. The WB as a qualitative BAB detection method had a given sensitivity of 97% and a specificity of 55%. There was a strong and significant correlation between high NAB titers (>500 neutralizing units [NU]) and titers obtained by all quantitative BAB assays. However, low to medium NAB titers (20-500 NU) did not significantly correlate with BAB titers. CONCLUSION: We conclude that the cELISA seems to be most suitable for NAB screening, but BAB titers cannot reliably predict NAB titers.     

Vasculitic mononeuritis multiplex in patient with Lyme disease

Lyme borreliosis is an infectious disease caused by the spirochete Borrelia burgdorferi. Neurological complications are frequently reported but the pathogenesis remains largely unknown. We report on a patient with positive borrelia antibodies at ELISA and immunoblotting and with histological and immunofluorescence study of peripheral nerve biopsy consistent with vasculitic neuropathy.

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Sommario La borreliosi di Lyme è una malattia infettiva causata dalla spirochete Borrelia burgdorferi. Le complicanze neurologiche sono segnalate con frequenza, ma la loro patogenesi rimane oscura. Noi presentiamo il caso di un paziente con positività di anticorpi al test ELISA e con studio istologico e di immunofluorescenza di una biopsia del nervo periferico che comprova l'esistenza di una neuropatia vasculitica.

     

Central nervous system abnormalities in Lyme neuroborreliosis

Intrathecal production of anti-Borrelia burgdorferi antibody occurs frequently in CNS Lyme, yet reliable diagnosis of neuroborreliosis is still considered difficult and controversial. Therefore, we assessed the utility of this measurement in 103 Lyme patients. Among 15 patients with Lyme meningoradiculitis and 41 controls, diagnostic specificity was 93% and sensitivity 87%. Application of this method permits the identification of a rare B burgdorferi-associated multifocal encephalitis (brain infection) and its differentiation from a milder encephalopathy, or confusional state; the latter may not require CNS bacterial invasion. The encephalitis involves white matter more often than gray; severity varies widely. Of six patients with this antibiotic-responsive encephalitis, five were positive for HLA DQw3(DQw7). We conclude that (1) measurement of intrathecal antibody production is a reliable indicator of CNS infection, (2) North American neuroborreliosis includes the same spectrum of neurologic dysfunction as described in Europe, and (3) HLA typing may be useful in furthering our understanding of severe CNS involvement.     

PCR detection of Borrelia burgdorferi DNA in cerebrospinal fluid of Lyme neuroborreliosis patients.

We used the polymerase chain reaction (PCR), a method useful in the detection of Borrelia burgdorferi in vitro, to evaluate CSF in patients thought to have neuroborreliosis. Nested pairs of oligonucleotide primers were designed to recognize the C-terminal region of B burgdorferi OspA. CSF samples were obtained from (1) patients with immunologic evidence of systemic B burgdorferi infection and clinical manifestations suggestive of CNS dysfunction, (2) seronegative patients with clinical disorders consistent with Lyme borreliosis, and (3) patient and contamination controls; all were analyzed in a blinded fashion. PCR detected B burgdorferi OspA DNA in CSF of (1) 10 of 11 patients with Lyme encephalopathy, (2) 28 of 37 patients with inflammatory CNS disease, (3) seven of seven seronegative patients with Lyme-compatible disorders, and (4) zero of 23 patient controls. Zero of 83 additional contamination controls were PCR-positive. In eight patients from whom we obtained CSF before and after parenteral antimicrobial therapy, PCR results invariably predicted clinical outcome accurately.     

Combination of transcranial sonography, olfactory testing, and MIBG myocardial scintigraphy as a diagnostic indicator for Parkinson's disease

Background: Appropriate diagnostic biomarkers are useful for improving speed and accuracy of a diagnosis. Substantia nigra (SN) hyperechogenicity visualized by transcranial sonography (TCS), olfactory dysfunction, and the reduced uptake of ¹²³I‐metaiodobenzylguanidine (MIBG) in myocardial scintigraphy have been suggested as potential biomarkers for the identification of Parkinson’s disease (PD). Objectives: To evaluate the diagnostic potential of these tests and to determine whether combining them increases their diagnostic power. Methods: Subjects were 44 patients with clinically diagnosed PD and 36 healthy controls. TCS of the SN, the odor stick identification test for Japanese (OSIT‐J), and MIBG myocardial scintigraphy were conducted. Results: Eleven patients with PD (25%) and four controls (11%) were excluded because of an insufficient acoustic temporal bone window in the TCS. Thus, 33 patients with PD and 32 healthy controls were finally included. The diagnostic sensitivity of TCS, OSIT‐J, and MIBG myocardial scintigraphy was 78.8%, 84.8%, and 60.6%, respectively. The specificity of TCS and OSIT‐J was 93.8% and 78.1%, respectively. The combination of TCS of the SN and OSIT‐J substantially increased the sensitivity to a sufficient level for discriminating patients with PD from controls. Conclusion: TCS of the SN and olfactory testing play complementary roles in increasing diagnostic power in PD. As both tests are easy to perform, noninvasive, and inexpensive, the combination of TCS of the SN and olfactory testing may contribute to early and accurate diagnosis of PD.     

Diagnostic accuracy of cardiac metaiodobenzylguanidine scintigraphy in Parkinson disease

Background and purpose:  To estimate the diagnostic accuracy of cardiac ¹²³I-metaiodobenzylguanidine (MIBG) scintigram for detection of Parkinson disease.
Methods:  A cross-sectional study with index test of MIBG scintigram and reference standard of U.K. Parkinson's Disease Brain Bank Criteria was performed in 403 patients. Ratio of cardiac-to-mediastinum MIBG accumulation was determined at 20 min (early H/M) and 4 h (late H/M). Area under the receiver-operator characteristic (ROC) curve, sensitivity and specificity in detecting Parkinson disease were analyzed. Accuracy was analyzed in a subgroup of patients with disease duration of 3 years or less.
Results:  Area under the ROC curve was 0.89 using either early or late H/M as a diagnostic marker (95% CI 0.85–0.92 for early H/M and 0.86–0.93 for late H/M). Sensitivity and specificity were 81.3% (76.1–85.8%) and 85.0% (77.7–90.6%) for early H/M and 84.3% (79.3–88.4%) and 89.5% (83.01–94.1%) for late H/M. In the subgroup with duration of 3 years or less, the ROC curve area, sensitivity, and specificity were 0.86 (0.79–0.92), 76.0% (64.8–85.1%), and 83.9% (71.7–92.4%) for early H/M and 0.85 (0.78–0.92), 73.3% (61.9–82.9%), and 87.5% (75.9–94.8%) for late H/M.
Conclusion:  Although diagnostic accuracy of cardiac MIBG scintigram is high, it is limited because of insufficient sensitivity in patients with short duration.     

Anti-basal ganglia antibodies in acute and persistent Sydenham's chorea

OBJECTIVE: To determine the sensitivity and specificity of methods to detect anti-basal ganglia antibodies (ABGA) in Sydenham's chorea (SC). BACKGROUND: SC is a delayed manifestation of group Abeta hemolytic streptococcal infection typically associated with rheumatic fever (RHF). SC is characterized by chorea and motor and neuropsychiatric symptoms. Patients with SC produce antibodies that cross-react with streptococcal, caudate, and subthalamic nuclei antigens detected using an immunofluorescent (IF) method with inconsistent reports of positivity. METHODS: The authors developed ELISA and Western immunoblotting (WB) methods to detect ABGA and compared these assays to IF. They investigated samples from patients with acute SC (n = 20), persistent SC (n = 16), control samples from RHF (n = 16), and healthy pediatric volunteers (n = 11). RESULTS: ABGA ELISA had a sensitivity of 95% and specificity of 93% in acute SC. Both WB and IF had a sensitivity of 100% and specificity of 93%. In the persistent SC group, ABGA sensitivity dropped to 69% using WB and to 63% using IF. Three common basal ganglia antigens were identified by WB in both acute and persistent SC (40 kDa [n = 15], 45 kDa [n = 15], and 60 kDa [n = 13]). There was no antibody reactivity to cerebellum, cerebral cortex, or myelin antigen preparations in any group. CONCLUSIONS: These results support the hypothesis that Syndenham's chorea is an autoantibody-mediated disorder. Western immunoblotting and immunofluorescence are the best methods for detecting anti-basal ganglia antibodies, and reactivity to basal ganglia antigens of 40, 45, and 60 kDa were commonly seen in both acute and persistent cases of SC.