Diagnostic tests for a new spirochete, Borrelia lonestari sp. nov.

Abstract

Bites from Amblyomma americanum, a hard tick, have been associated with a Lyme disease-like illness in the southeastern and south-central United States. Present in 2% of ticks collected in four states were uncultivable spirochetes. Through use of the polymerase chain reaction, partial sequences of the flagellin and 16s rRNA genes of microorganisms from Texas and New Jersey were obtained. The sequences showed that the spirochete was a Borrelia sp. but distinct from other known members of this genus, including B. burgdorferi, the agent of Lyme disease. Species-specific differences in the sequences of the flagellin protein, the flagellin gene and the 16s rRNA gene between the new Borrelia species and previously known species provide compositions and methods for assay for determining the presence of this new spirochete, or for providing evidence of past or present infection by this spirochete in animal reservoirs and humans.

Description

[0001] The government owns rights in the present invention pursuant to grant number AI24424 from the National Institutes of Health.FIELD OF THE INVENTION [0002] The present invention relates generally to the fields of infection and disease. More particularly, it concerns the identification of a new spirochete carried by the hard tick, Amblyomma americanum, found by the present inventor to be associated with a Lyme disease-like illness in the southeastern and south-central United States. Most particularly, the invention provides compositions, methods, and kits for the identification of the new spirochete for diagnostic purposes. DESCRIPTION OF THE RELATED ART [0003] A paradox about Lyme disease is the report of this tick-borne infection from areas in which transmission of the etiologic agent, B. burgdorferi, has not been documented (Sigal et al., 1991; Barbour et al., 1993). This phenomenon has been reported from Georgia and Missouri, but may be common in other parts of the southeast...

AI Technical Summary

Benefits of technology

[0028] A fusion protein or peptide comprising a segment of SEQ ID NO:2 having at least one B. lonestari sp. nov.-specific amino acid or species-specific combination of amino acids of Table 1 is also an aspect of the present invention. The fusion protein preferably comprises SEQ ID NO:26, however, one skilled in the art, in light of the present disclosure, would be able to construct a number of different fusion proteins from a variety of vectors and the B. lonestari sp. nov. DNA sequences provided herein. It will also be understood that amino acid and nucleic acid sequences may include additional residues, such as additional N— or C-terminal amino acids, and yet still be essentially as set forth in one of the sequences disclosed herein, so long as the sequence meets the criteria set forth above. Segments of the flagellin gene may be cloned next to N— and / or C-terminal sequences of genes for other proteins, such as, β-galactosidase or maltose binding protein. A signal peptide that may allow better expression may be optionally included in the fusion protein. It is not necessary that the flagellin protein be transported, however, the signal peptide may help to prevent protease digestion.

Problems solved by technology

Although there is much known about Lyme disease, there are currently no means of identification of the new spirochete associated with the aforedescribed Lyme disease-like pathology and further, no means of diagnosis of infection, compositions for clinical tests, or laboratory assays for diagnosing a patient exhibiting Lyme disease-like symptoms but testing negative for Lyme disease.

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