Method, kits and materials for detection of Lyme disease
View Patent ↗Disclosed herein are method, materials and kits for detection of B. Burgdorferi infection or determining stage of Lyme disease in a subject. Exemplified is a method of diagnosing an infection in a subject, the method involving exposing a biological sample from the subject to a capture substrate under conditions for an infection marker in said biological sample to associate with the capture substrate to form a capture complex; associating said capture complex with a marker complex, said marker complex comprising an oligonucleotide; and amplifying said oligonucleotide of marker complex associated with said capture complex to produce an amplification signal; wherein an amplification signal above a predetermined signal threshold indicates that said subject is infected.
1. A method of detecting a Borrelia species infection marker in a human or non-human mammal subject, the method comprising: exposing a biological sample from the subject to a capture substrate comprising a recombinant antigen that comprises a fusion protein of SEQ ID NO: 1 under conditions for an infection marker in said biological sample to associate with the capture substrate to form a capture complex; associating said capture complex with a marker complex, said marker complex comprising an oligonucleotide; amplifying said oligonucleotide of marker complex associated with said capture complex to produce an amplification signal and detecting presence of the infection marker in the sample based on if the amplification signal is above a predetermined threshold.
2. The method of claim 1 wherein said capture substrate comprises a solid support with the fusion protein bound thereto.
3. The method of claim 2 , wherein said solid support is a bead.
4. The method of claim 1 , wherein said Borrelia species is Borrelia burgdorferi, Borrelia afzelli and/or Borrelia garinii.
5. The method of claim 1 , wherein said at least one recombinant antigen is produced with a tag that is subsequently cleaved.
6. The method of claim 5 , wherein said tag is a glutathione-S-transferase (GST) tag, a hemagglutinin, or C-Myc, or combinations thereof.
7. The method of claim 1 , wherein said infection marker is a primary host antibody that binds to a Lyme disease Borrelia species or a Lyme disease Borrelia epitope comprising antigen.
8. The method of claim 1 , wherein the amplification signal is fluorescence.
9. The method of claim 8 , wherein the predetermined signal threshold is greater than or equal to three standard deviations above a mean background signal.