A Fully Automated Multiplex Assay for Diagnosis of Lyme Disease with High Specificity and Improved Early Sensitivity

Abstract

Lyme borreliosis is a tick-borne disease caused by the Borrelia burgdorferi sensu lato complex. Bio-Rad Laboratories has developed a fully automated multiplex bead-based assay for the detection of IgM and IgG antibodies to B. burgdorferi. The BioPlex 2200 Lyme Total assay exhibits an improved rate of seropositivity in patients with early Lyme infection. Asymptomatic subjects from endemic and nonendemic origins demonstrated a seroreactivity rate of approximately 4% that was similar to other commercial assays evaluated in this study. Coupled to this result was the observation that the Lyme Total assay retained a high first-tier specificity of 96% while demonstrating a relatively high sensitivity of 91% among a well-characterized CDC Premarketing Lyme serum panel. The Lyme Total assay also performs well under a modified two-tier algorithm (sensitivity, 84.4 to 88.9%; specificity, 98.4 to 99.5%). Furthermore, the new assay is able to readily detect early Lyme infection in patient samples from outside North America.

The Performance of Nine Commercial Serological Screening Assays for the Diagnosis of Lyme Borreliosis: a Multicenter Modified Two-Gate Design Study

“…In contrast, IgM positivity rates were much higher and might be explained by cross-reactivity with antibodies raised against heterologous bacteria and autoimmune diseases or by polyclonal B-cell activation ( 16 ). The higher positivity rates among the cross-reactivity controls confirmed what has been published before ( 6 , 7 , 11 , 17 ) and underlined that positive IgM serology results should be interpreted with care, especially when clinical symptoms are unclear.…”

A refined human linear B cell epitope map of Outer surface protein C (OspC) from the Lyme disease spirochete,Borreliella burgdorferi

“…The C10 peptide corresponds to the stretch of ∼10 highly conserved amino acids at C-terminus of OspC, first recognized as an immunodominant epitope in neuroborreliosis patients (52). Since then, the C10 peptide and derivatives have been integrated into numerous diagnostic assays, including the Zeus VlseE1/C10 ELISA (25, 27, 28). Dolange and colleagues recently proposed the use of high affinity C10-specific monoclonal antibodies as tools to detect circulating levels of B. burgdorferi antigens (54).…”

“…OspC is implicated in facilitating spirochete egress from the tick during the course of a blood meal, enabling survival in the early stages of mammalian skin infection, and modulating transmigration across vascular walls (16)(17)(18)(19)(20)(21)(22)(23). The highly immunogenic nature of OspC has lent itself to applications in Lyme disease serology-based diagnostics, including widely used commercial tests (24,25). For example, the peptide derived from the conserved C-terminal 10 amino acids of OspC (peptide "C10") is a component of the FDA approved diagnostic test Borrelia VlsE1/pepC10 from Zeus Scientific (Branchburg, NJ) (26)(27)(28)(29)(30).…”

Immunoblot Criteria for Diagnosis of Lyme Disease: A Comparison of CDC Criteria to Alternative Interpretive Approaches

“…The breadth of the immune response provided by immunoblotting may provide a window to help answer the latter question, but, as recognized by Liu et al [30], large prospective studies would be required to optimize disease-stage specific diagnosis using immunoblot techniques. Although immunoblot responses can be quantified using densitometry and multivariate algorithms [41,76], multiplex technologies for antibody detection using microsphere and plasmonic biochip techniques offer a broader dynamic range than immunoblotting and greater reproducibility [77][78][79]. The availability of highly sensitive and reproducible multiplex techniques argues that prospective studies using these newer technologies might be more fruitful than employing an older methodology.…”