Early and accurate differential diagnosis of Lyme and other tick-borne diseases by agglutination-PCR

This proposal focuses on the development of a highly sensitive and specific multiplex immune marker quantification tool for monitoring host immunoglobulin responses against Lyme and other blacklegged tick borne pathogens using Antibody Detection by Agglutionation PCRADAPtechnologyLyme diseaseLDis often missed or unrecognized and if not treated definitively during early infectionpatients can suffer severe and often irreversible consequencesIn the USinfection with the Lyme disease pathogenBorrelia burgdorferiis increasingly prevalentaffecting overyrUnlike other bacterial infectionsdirect detection of LD bacterial genomic materials and antigens is not useful in early diagnosis due to the extreme scarcity of LD bacteria in the circulationThe two tier serological testing algorithmfirst tier enzyme immunoassay followed by a second tier Western blotis still the mainstay of Lyme testingHoweverthis two tier algorithm identifies less thanof early LD infectionssignaling an urgent and compelling need for a better diagnostic assayIn additionticks transmitting LD can also contain other pathogens causing debilitating co infections requiring accurate differential diagnosisIn particularBorrelia miyamotoi inducedRelapsing Fever and Babesiosis are two common diseases requiring different antimicrobial managementThis project proposes to adapt and employ our novel ADAP technology that harnesses the exponential amplification attributes of PCR to achieve highly sensitive detection of host antibody immune responsesADAPandapos s DNA barcoding will be used to enable simultaneous multiplex quantification of immune markers against several LymeRelapsing Fever and Babesiosis antigens inL of samplesThe proposed multiplex ADAP tick borne disease assayandapos s enhanced sensitivity can enable much earlier disease detection during the critical window period for best outcomes and reveal possible co infection s for most appropriate treatmentIn Specific Aimwe will express and purify several Lyme recombinant proteins that are not currently commercially availableThenwe will synthesize corresponding antigen DNA conjugate pairs for a ADAP Lyme assayFinallywe will optimize and validate the assayandapos s performance using well annotated patient samples from our Co Investigatorandapos s sites at Columbia University in New YorkTulane in New Orleans and from the CDCNotablywe present a plan to rigorously investigate and mitigate potential cross reactivity with other diseasessuch as syphilis and rheumatoid arthritisIn Specific Aimwe will further express and purify antigens for Relapsing Fever and Babesiosis and then synthesize and validate corresponding antigen DNA conjugates using patient samplesIf successfulconjugates from Aimandwill be combined into a single multiplex tick borne disease assay for blacklegged tick transmitted pathogensThe positive outcome of this project will be a comprehensive assay able to identify patients during the earliestmost valuable treatment windowoffering an unprecedented opportunity to substantially improve outcomes and reduce the profound personal and societal burden of chronic disease Public Health Statement The growing menace of Lyme diseaseLDtransmitted via bites of infected ticks affects overindividuals in the US annually and if not detected and differentiated from other tick borne diseases and treated definitively during early infection can cause irreversible debilitating consequencesEnable Biosciences proposes to use its novel ADAP technology to develop and optimize a highly sensitive and very specific assay capable of identifying and differentiating Lyme from other tick borne infections at the earliest possible stage enabling administration of effective targeted therapy that can dramatically improve patient outcomes and reduce severe chronic disease