SBIR-STTR Award

There is no validated assay to reliably detect the early onset of the Lyme disease infection at the time when antibiotic therapy has the best chance of a cure without sequelae. Diagnosis is often difficult because most symptoms are often nebulous, with the telltale "bulls-eye" rash only appearing and clearly recognizable in approximately half of cases. The standard types of infectious diagnostic tests - both culture and serology - require >3 weeks for a positive result after the onset of infection with Lyme disease. This introduces a significant delay in treatment. While PCR for B. burgdorferi DNA in the blood has potential for a rapid, accurate result, past tests have not had sufficient sensitivity because f few bacteria and low copy numbers of DNA in the blood. We have developed an approach that "enlarges the target", substantially increasing the sensitivity of PCR for B. burgdorferi. Our preliminary data show that we can accomplish this by: 1) obtaining a larger sample of blood than usual (10X increase in sensitivity), 2) selectively multiplying the target by using Borrelia-directed primers prior to PCR (200X increase), and 3) probing for the B. burgdorferi on a multi-loci PCR platform (8X increase). To assure reliability and acceptance of a new test based on this approach, we have formally consulted with the FDA from the beginning, and obtained a pre-IDE approval with guidance as to what we need to do for a final cleared test. In this proposal we will test blood samples (from eventually confirmed Lyme cases) with our PCR-based assay at the time of patient presents to prove that it can detect the B. burgdorferi infection earlier and with greater sensitivity specificity than two- tiered serology taken at the same time. Specificity will be assured by testing endemic area controls without Lyme disease. If successful, as our Preliminary and published data suggest, we will follow with a Phase II proposal that analyzes greater number of Lyme disease and control samples to meet the full number for FDA clearance. Our goal is deliver a functional diagnostic assay for Early Onset Lyme Disease that has superior sensitivity and specificity compared to two-tiered serology. The test will permit immediate diagnosis well before seroconversion or cultures become positive so treatment can be started right away.

Public Health Relevance Statement:


Public Health Relevance:
The ability to accurately identify Lyme disease in the susceptible people will be important in management strategy of patients exposed or bitten by ticks. It will help reduce rising costs to the patient and the public health system.

Project Terms:
Antibiotic Therapy; Antibiotics; Antibodies; Area; Bacteria; base; Biological Assay; Bite; Blood; Blood specimen; Blood Tests; Borrelia; Borrelia burgdorferi; burden of illness; Centers for Disease Control and Prevention (U.S.); Clinical; Clinical assessments; Communicable Diseases; Communities; Confusion; Consult; cost; Data; Deposition; design; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; disorder control; DNA; DNA copy number; early onset; erythema migrans; Exanthema; Eye; Feeling; flu; Goals; Health Care Costs; Health system; Heart; Hour; improved; Individual; Infection; Joints; Letters; Lower Organism; Lyme Disease; Medical; meetings; Methods; Microbe; Molecular Diagnostic Testing; Nervous system structure; Patients; Phase; prevent; Procedures; public health medicine (field); public health relevance; Publishing; Sampling; Sensitivity and Specificity; Serologic tests; skin lesion; Specificity; Specimen; Symptoms; Testing; Textbooks; Ticks; Time; Tissues; tool; Visit; West Nile virus; Work

There is no validated assay to reliably detect the early onset of the Lyme disease infection at the time when antibiotic therapy has the best chance of a cure without sequelae. Diagnosis is often difficult because most symptoms are often nebulous, with the telltale “bulls-eye” rash only appearing and clearly recognizable in approximately half of cases. The standard types of infectious diagnostic tests – both culture and serology – require >3 weeks for a positive result after the onset of infection with Lyme disease and serology only indicates exposure but not active infection. This introduces a significant delay in treatment. While PCR for B. burgdorferi DNA in the blood has potential for a rapid, accurate result, past tests have not had sufficient sensitivity because of too few bacteria and low copy numbers of DNA in the blood. We have developed an approach that “enlarges the targetability”, by 160,000 fold, thereby increasing the sensitivity of PCR test for B. burgdorferi. Our data show we are successfully accomplishing this by: using a larger amount of blood to start (10 fold); then selectively multiplying the target by using Borrelia-directed primers prior to PCR (2000 fold); and then probing the product for the B. burgdorferi on a multi-loci PCR platform (8 fold). To assure reliability and acceptance of a new test based on this approach, we formally consulted with the FDA from the beginning. We obtained a Qsub (pre-IDE) approval with guidance as to what we need for a cleared test. During this proposal we will continue to test blood samples (from eventually confirmed Lyme cases and control subjects) with our PCR-based assay at the time the patient presents to the doctor in order to prove the assay can detect the B. burgdorferi infection, earlier and with greater sensitivity, than the FDA benchmark of two-tiered serology taken at the same time. If successful, at the number agreed on with the FDA, this Phase II should result in a test that has FDA clearance. Our goal is deliver a functional and affordable diagnostic assay for Early Onset Lyme Disease that has superior sensitivity and specificity compared to two-tiered serology. The test will permit immediate diagnosis well before seroconversion or cultures become positive so treatment can be started right away.

Public Health Relevance Statement:
The ability to accurately identify Lyme disease in the susceptible people will be important in management strategy of patients exposed or bitten by ticks. It will help reduce rising costs to the patient and the public health system.

Project Terms:
Antibiotic Therapy; Antibiotics; Antibodies; Antibody Response; Area; Bacteria; base; Benchmarking; Biological; Biological Assay; Blood; Blood specimen; Blood Tests; Blood Volume; Borrelia; Borrelia burgdorferi; burden of illness; case control; Centers for Disease Control and Prevention (U.S.); Clinical; clinically relevant; common symptom; Communicable Diseases; Consult; cost; Data; Deposition; Detection; Development; Diagnosis; Diagnostic; diagnostic assay; Diagnostic tests; disabling symptom; disorder control; DNA; DNA copy number; Early Diagnosis; early onset; Economics; Enzyme-Linked Immunosorbent Assay; erythema migrans; Exanthema; Eye; Feeling; flu; Goals; Health system; Heart; Hour; impression; Infection; Laboratories; Left; Lyme Disease; Methods; Microbe; Molecular Diagnostic Testing; Nervous system structure; novel strategies; nucleic acid detection; pathogen; Patients; Phase; prevent; Public Health; Publishing; Research Design; Sampling; Sensitivity and Specificity; seroconversion; Serologic tests; skin lesion; Sore Throat; Specificity; Spottings; Symptoms; Technology; Testing; Ticks; Time; Tissues; United States; Urinary tract infection; West Nile virus; Western Blotting; Whole Blood; Work