Hunfeld Klaus-Peter, Ernst Manfred, Zachary Pierre, Jaulhac Benoit, Sonneborn Hans-Hermann, Brade Volker
Institute of Medical Microbiology, University Hospital of Frankfurt/Main, Germany.
Wien Klin Wochenschr. 2002 Jul 31;114(13-14):580-5.
The use of recombinant proteins for serologic testing represents a modern approach for the improved laboratory diagnosis of Lyme disease (LD). The aim of the present study was to develop and evaluate a new recombinant ELISA (RE) for the detection of specific IgG and IgM antibodies against Borrelia burgdorferi.
The RE (Biotest AG, Dreieich, Germany) uses mixtures of recombinant p100, OspC, p18 of Borrelia afzelii and a fusion protein of recombinant internal fragments of the flagellum protein (p41) of Borrelia garinii strain PBi and Borrelia afzelii strain PKo. Serologic testing was performed on a commercially available ELISA processor without pre-absorption of sera. The sensitivity of the RE was determined by testing 226 sera obtained from patients suffering from Lyme disease (stage I: n = 148, stage II: n = 35, stage III: n = 43). Specificity of the RE was evaluated in 1107 sera from healthy blood donors and 275 sera from patients with other infectious diseases or autoimmune illnesses (leptospirosis: n = 53, syphilis: n = 70, toxoplasmosis: n = 60, herpes simplex virus: n = 30, HIV: n = 30; rheumatoid-factor positive: n = 32). In addition, 394 routine samples were prospectively tested in comparison with a well established whole-cell lysate extract ELISA (ENZYGNOST Borreliosis, DadeBehring, Germany) for relative sensitivity and specificity.
Overall specificity, determined in 1107 healthy blood donors and 275 sera from patients with other diseases, was 94% for IgG and 91% for IgM. The overall sensitivities in 226 sera obtained from patients suffering from different stages of LD were 67-95%. Moreover, as revealed by prospective testing of 394 routine samples, the relative sensitivity of the RE in comparison with an established whole-cell lysate extract ELISA in the detection of seropositive samples was 81.1% for IgG and 86.5% for IgM with a relative specificity of 98.5% and 93% respectively. The ELISAs showed an overall agreement of 97% for IgG and 92.4% for IgM test results.
The RE proved to be a reliable and specific screening test in the routine serodiagnosis of LD. In addition, the RE is easy to perform and requires no pre-absorption.
使用重组蛋白进行血清学检测是改进莱姆病（LD）实验室诊断的现代方法。本研究的目的是开发并评估一种新型重组酶联免疫吸附测定法（RE），用于检测抗伯氏疏螺旋体的特异性IgG和IgM抗体。
该RE（德国德赖艾希市比奥泰克公司）使用重组p100、OspC、阿氏疏螺旋体p18的混合物，以及加氏疏螺旋体菌株PBi和阿氏疏螺旋体菌株PKo鞭毛蛋白（p41）重组内部片段的融合蛋白。血清学检测在市售酶联免疫吸附测定仪上进行，血清无需预先吸收。通过检测226份莱姆病患者的血清（I期：n = 148，II期：n = 35，III期：n = 43）来确定RE的敏感性。在1107份健康献血者的血清和275份患有其他传染病或自身免疫性疾病患者的血清（钩端螺旋体病：n = 53，梅毒：n = 70，弓形虫病：n = 60，单纯疱疹病毒：n = 30，HIV：n = 30；类风湿因子阳性：n = 32）中评估RE的特异性。此外，与一种成熟的全细胞裂解物酶联免疫吸附测定法（德国达德拜林公司的ENZYGNOST莱姆病检测试剂盒）相比，对394份常规样本进行前瞻性检测，以确定其相对敏感性和特异性。
在1107份健康献血者血清和275份其他疾病患者血清中测定的总体特异性，IgG为94%，IgM为91%。在226份不同阶段莱姆病患者的血清中测定的总体敏感性为67 - 95%。此外，通过对394份常规样本的前瞻性检测发现，与成熟的全细胞裂解物酶联免疫吸附测定法相比，RE在检测血清阳性样本时，IgG的相对敏感性为81.1%，IgM为86.5%，相对特异性分别为98.5%和93%。两种酶联免疫吸附测定法的IgG检测结果总体一致性为97%，IgM为92.4%。
在莱姆病的常规血清学诊断中，RE被证明是一种可靠且特异的筛查试验。此外，该RE操作简便，无需预先吸收。
