Point-of-care serodiagnostic test for early-stage Lyme disease using a multiplexed paper-based immunoassay and machine learning

Abstract

Caused by the tick-borne spirochete, Borrelia burgdorferi, Lyme disease (LD) is the most common vector-borne infectious disease in North America and Europe. Though timely diagnosis and treatment are effective in preventing disease progression, current tests are insensitive in early-stage LD, with a sensitivity <50%. Additionally, the serological testing currently recommended by the US Center for Disease Control has high costs (>$400/test) and extended sample-to-answer timelines (>24 hours). To address these challenges, we created a cost-effective and rapid point-of-care (POC) test for early-stage LD that assays for antibodies specific to seven Borrelia antigens and a synthetic peptide in a paper-based multiplexed vertical flow assay (xVFA). We trained a deep learning-based diagnostic algorithm to select an optimal subset of antigen/peptide targets, and then blindly-tested our xVFA using human samples (N(+) = 42, N(-)= 54), achieving an area-under-the-curve (AUC), sensitivity, and specificity of 0.950, 90.5%, and 87.0% respectively, outperforming previous LD POC tests. With batch-specific standardization and threshold tuning, the specificity of our blind-testing performance improved to 96.3%, with an AUC and sensitivity of 0.963 and 85.7%, respectively.

Comparative Cost and Effectiveness of a New Algorithm for Early Lyme Disease Diagnosis: Evaluation in US, Germany, and Italy

“…[37][38][39][40] From an economic standpoint, the expected reduction in disease burden is consistent with the findings of previous research, highlighting the economic potential of late LD stages prevention by improved diagnostic accuracy, or by vaccination. [19][20][21]24,25,28,31,41,42 We point out that indirect costs have not been incorporated in the model, for two main reasons: firstly, the adopted cost perspective is the one of the third-party payers that will ultimately have to decide upon the funding of the new technology, and secondly the limited data availability with estimates available only for the Netherlands, not for the countries evaluated. 21,31 It is conceivable that savings with the most efficient technology would increase in case productivity losses due to the disease and its management would be factored into the analysis.…”

Lyme Disease Biosensors: A Potential Solution to a Diagnostic Dilemma

“…More recently, Joung et al (2020) improved upon their previous design by incorporating four additional antigens (DbpB, Crasp1, P35, Erpd/Arp37) and a synthetic peptide (Mod-C6) into the sensing membrane [ 42 ]. In addition, they used deep learning to train a neural network to determine an optimal diagnostic algorithm.…”

“…Likewise, the MTT approach demonstrates only slightly improved sensitivity, at approximately 50%, for patients with early Lyme disease [ 18 ]. Some of the biosensors presented here ( Table 1 ) report sensitivities for early Lyme that exceed the STT/MTT approaches and have the potential to develop into a much-needed alternative [ 32 , 33 , 42 ]. However, some biosensors also demonstrate reduced specificity in comparison to the two-tiered approaches.…”

“…In addition to their initial low sensitivity, the STT and MTT approaches are also quite expensive, both in analysis time (>24 h) and cost (>$400 per test) [ 42 ]. 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. These biosensors represent a substantial advance in the improvement of Lyme diagnostics and open the door to new inventions and innovations for both the diagnosis of Lyme disease and other infectious agents.…”