title: Development and laboratory evaluation of a new recombinant ELISA for the serodiagnosis of Lyme disease.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute of Medical Microbiology, University Hospital of Frankfurt/Main, Germany. K.Hunfeld@em.uni-frankfurt.de
authors.forename: Klaus-Peter
authors.initials: KP
authors.lastname: Hunfeld
authors.forename: Manfred
authors.initials: M
authors.lastname: Ernst
authors.forename: Pierre
authors.initials: P
authors.lastname: Zachary
authors.forename: Benoit
authors.initials: B
authors.lastname: Jaulhac
authors.forename: Hans-Hermann
authors.initials: HH
authors.lastname: Sonneborn
authors.forename: Volker
authors.initials: V
authors.lastname: Brade
mesh_terms: D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood*; D001426:Bacterial Proteins / Q000276:immunology; D015748:Borrelia burgdorferi Group / Q000276:immunology*; D003429:Cross Reactions; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015787:Erythema Chronicum Migrans / Q000175:diagnosis / Q000276:immunology; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D007075:Immunoglobulin M / Q000097:blood*; D008193:Lyme Disease / Q000175:diagnosis* / Q000276:immunology; D008297:Male; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011994:Recombinant Proteins