subject id: 103 visit number (1 = baseline): 1 Sex: F elevated liver enzyme [ast, alk, or alk-phos at pre-treatment visit (v1). 0 = no, 1 = yes]: N multiple em (y = multiple erythema migrans lesions, n = single erythema migrans lesion): N/A illness duration (days of illness prior to treatment beginning): N/A # of symptoms: N/A fatigue severity [range from 9-63] scores ≥36 are considered "high"]: 25 ptlds status [ptlds status 0 = returned to health, 1 = symptoms only, 2 = ptlds]: N/A mcgill pain total score (range 0-45): 5
Extracted molecule
protein
Extraction protocol
Serum was isolated from coagulated blood using standard protocols. All sera were frozen and aliquoted within four hours of sampling.
Label
PE
Label protocol
Spectrally-distinct polystyrene beads with magnetic cores were covalently coupled to capture antibodies by the manufacturer. All assays were performed using 96-well flat-bottom plates and a Bio-Plex Pro II magnetic wash station. Standards, quality controls and experimental sera were diluted in provided diluents and then incubated with the respective multiplex bead mix according to manufacturer protocol.
Hybridization protocol
Incubation with the appropriate biotinylated detection antibody cocktail was performed as outlined in the protocol, followed by incubation with streptavidin-PE. Following the final wash step, beads were re-suspended in the designated buffer.
Scan protocol
Plates were analyzed using a Luminex 200 (Luminex Corporation, Austin, TX, USA) in combination with Bio-Plex Manager 5.0 software (Bio-Rad Laboratories, Hercules, CA, USA).
Data processing
Serum concentrations were interpolated from standard curves for each respective analyte in Bio-Plex Manager 5.0 software. Values out of range below the standard curve were set to zero, while any value out of range above the standard curve was set to 4x the highest observed standard concentration. For descriptive analyses of the multiplex biomarker data, data were analyzed using the log of the “ratio to cohort average”. This value was computed for each analyte by first setting any value less than 1 pg/mL to 1 pg/mL and then calculating the log base 2 of [(observed value)/(average value in the cohort)].
