Development of a multiplex and sensitive lateral flow immunoassay for the diagnosis of periprosthetic joint infection

Abstract

The diagnosis of periprosthetic joint infection (PJI) remains a challenge. However, recent studies showed that synovial fluid biomarkers have demonstrated greater diagnostic accuracy than the currently used PJI diagnostic tests. In many diagnostic tests, combining several biomarkers into panels is critical for improving diagnostic efficiency, enhancing the diagnostic precision for specific diseases, and reducing cost. In this study, we prove that combining alpha-defensin and C-reactive protein (CRP) as biomarkers possesses the potential to provide accurate PJI diagnosis. To further verify the result, we developed a multi-target lateral flow immunoassay strip (msLFIA) with staking pad design to obtain on-site rapid response for clinical diagnosis of PJI. A total of 10 synovial fluid samples were tested using the msLFIA, and the results showed that the combined measurements of synovial fluid alpha-defensin and CRP levels were consistent with those obtained from a commercial enzyme-linked immunosorbent assay kit. In addition, we developed a multi-target lateral flow immunoassay strip (msLFIA) with staking pad design to obtain on-site rapid response for clinical diagnosis of PJI, which the multi-target design is used to increase specificity and the stacking pad design is to enhance detection sensitivity. As a result, the turnaround time of the highly sensitive test can be limited from several hours to 20 min. We expect that the developed msLFIA possesses the potential for routine monitoring of PJI as a convenient, low-cost, rapid and easy to use detection device for PJI.

Interconnectable solid-liquid protein extraction unit and chip-based dilution for multiplexed consumer immunodiagnostics

“…Even with ULOC-enabled sample dilution (DF x 20), high-concentration LFIA effects are observed, affirming the necessity of dilution to avoid false-negative results. The majority of LFIA developers recommend an assay duration of up to 20 min to allow the signal to reach full stability and sensitivity [ 8 ]. However, we found that a 15 min duration was suitable for allowing signals to develop for all concentrations, without neglecting more concentrated samples.…”

“…To date, the lateral flow immunoassay (LFIA) is the most successful application of consumer diagnostics [ 6 ]. Combining LFIAs with smartphones as optical detectors allow for ‘on-the-go’ decentralized screening [ 7 ] and smartphones can even provide semi-quantitative results by calibrating test and control line intensity values toward a particular antigen concentration [ 8 ].…”

Recent Trends in Nanomaterial-Based Biosensors for Point-of-Care Testing

“…Additionally, multiple test lines can be used for simultaneous detection of different targets (Song et al, 2014). For instance, Tsai et al developed a multiplex and sensitive lateral flow assay for simultaneous detection of alpha-defensin and C-reactive protein (Tsai et al, 2019).…”

Application of leukocyte esterase strip test in the screening of periprosthetic joint infections and prospects of high-precision strips

“…The existing methods for infection diagnosis mainly involved microbial culture [31] and non-culture methods, including bacterial staining, white blood cell counting, antibody and antigen immunoassay, and next generation sequencing (mNGS) [32][33][34][35][36][37]. These methods are of high diagnostic value but require complex specialized equipment, special site environment and professional staff.…”