A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

Abstract

Lyme disease (LD) diagnosis using the current two-tier algorithm is constrained by low sensitivity for early-stage infection and ambiguity in determining treatment response. We recently developed a protein microarray biochip that measures diagnostic serum antibody targets using grating-coupled fluorescent plasmonics (GC-FP) technology. This strategy requires microliters of blood serum to enable multiplexed biomarker screening on a compact surface and generates quantitative results that can be further processed for diagnostic scoring. The GC-FP biochip was used to detect serum antibodies in patients with active and convalescent LD, as well as various negative controls. We hypothesized that the quantitative, high-sensitivity attributes of the GC-FP approach permit: 1) screening of antibody targets predictive for LD status, and 2) development a diagnostic algorithm that is more sensitive, specific, and informative than the standard ELISA and Western blot assays. Notably, our findings led to a diagnostic algorithm that may be more sensitive than the current standard for detecting early LD, while maintaining 100% specificity. We further show that analysis of relative antibody levels to predict disease status, such as in acute and convalescent stages of infection, is possible with a highly sensitive and quantitative platform like GC-FP. The results from this study add to the urgent conversation regarding better diagnostic strategies and more effective treatment for patients affected by tick-borne disease.

Multiplexed Detection and Quantification of Human Antibody Response to COVID-19 Infection Using a Plasmon Enhanced Biosensor Platform

“…For dried blood spot samples, ROC analysis yielded 100% specificity and variable sensitivity when the following GC-FP detection ratio thresholds were met: S1 = 0.855, S1S2 = 1.12, and Nuc = 0.98. AUC values ranged from 0.85 to 0.92, all of which are well above the cut-off value of 0.7 that we have successfully used for GC-FP based Lyme disease diagnosis (Chou et al 2020). When these GC-FP detection ratio thresholds were exceeded (to maintain 100% specificity) assay sensitivity was relatively low for S1 and S1S2 antigens (66.7% for both) but increased to 86.7% for the nucleocapsid antigen (Nuc).…”

“…for Lyme disease testing (Chou et al 2020). Blood serum was prepared after venipuncture and collection in serum collection tubes by centrifugation at 1,000 rpm for 15 min, followed by removal of the supernatant (serum).…”

Lyme Disease Biosensors: A Potential Solution to a Diagnostic Dilemma

“…More recently, Chou et al (2020) presented a microfluidic protein microarray biochip ( Figure 4 ) for the detection of antibodies against 16 different B. burgdorferi antigens [ 47 ]. Similar to the previous biochip, Borrelia antigens were immobilized onto a microchip—this time using a robotic microarrayer.…”

“…A rapid, inexpensive, and portable biosensor would prove to be an extremely beneficial tool, especially for rural areas where access to healthcare is limited. Several of the biosensors reviewed here have the potential to develop [ 37 , 47 ], or have already developed [ 39 ], into commercial Lyme sensors that approach or exceed the sensitivity and specificity of the STT and MTT methods. As well, some of the biosensors presented here utilize methods that are relatively inexpensive [ 39 , 41 , 42 ] in comparison to the STT/MTT approaches.…”

“… Microfluidic protein biochip showing ( A ) biochip device with overlying microfluidic window ( B ) illustration of antigen-antibody binding process with Surface Plasmon Resonance (SPR)-enhanced fluorescence ( C ) fluorescent image of an antibody-bound biochip. Reprinted from ref [ 47 ], which is open access under the Creative Commons Attribution 4.0 International license. …”