Background

Borrelia burgdorferi is an infectious spirocheate bacterium that is the causative agent of Lyme disease. B. burgdorferi infections cause a wide range of morbidity and mortality in the continental United States. The current suite of testing methods to diagnose and/or detect the presence of B. burgdorferi in patient samples revolves around three basic testing modalities. These include microscopy, serologic, and molecular diagnostic based approaches. Microscopic methods typically rely on visualizing blood during an early active phase of infection or in biopsies of affected skin displaying the characteristic erythema migrans rash, however detection of blood-borne spirocheates is variable and depends on the level of infection, therefore this method is limited due to the level and presentation of focal infection sites. Serology based assays are aimed at the detection of hosts development of specific anti-Borrelia antibodies (IgG or IgM) to specific Borrelia antigens. However, it is of general consensus among infectious disease specialists that not all Borrelia infections result in a predictable antigenic response and limiting detection methods to a small subset of antigenic proteins is likely resulting in unexpected false negatives. This is further complicated in that antibody production may be limited in certain patient populations due to various unrelated health condition or medication. Molecular based detection methods rely on the direct detection of the B. burgdorferi genome and isolating or visualizing DNA sequences diagnostic for that organism. This approach has variable results depending on the way the testing method is designed and the regions of DNA that are targeted for detection. The recent trend in diagnostics is to provide a complete detection package that includes serologic, microscopic and molecular, particularly PCR based, assays aimed at the specific and sensitive detection of Borrelia spp. in patient samples.

PCR Test Literature Survey

The first documented example of PCR based detection of B. burgdorferi was conducted in Maryland early in 1990 [1]. The application of PCR for the specific detection of Borrelia spp. in humans has rapidly expanded and has continued as a robust area of scientific inquiry over the last 10 years [2-66]. The use of PCR as a detection technology has been studied specifically to aid in the discrimination of varying types of clinical symptoms, their presentation, and subsequent identification [7, 12, 15, 18, 25, 30, 34, 41, 43, 48, 52, 56, 60, 62, 64, 65, 67-77]. The use of Borrelia spp. PCR technology is further bolstered by a suite of comparison studies detailing the use, relative strengths, and weaknesses when compared to varying clinical detection technologies [11, 15-17, 23, 24, 26, 32-34, 36-38, 42, 45, 51, 53-55, 57, 59, 61, 65, 66, 77, 78]. Borrelia spp. PCR has also been used to follow treatment for Lyme disease [40, 64] and following through with long term chronic symptoms of Lyme disease [18, 59, 76, 79]. Finally, Borrelia spp. PCR has been used to study various strains, genospecies, and strains of Borrelia which originate from infected humans [4, 7, 9, 13, 46, 47, 50, 58, 74, 80]. Borrelia spp. PCR is not only restricted to detection of the organism in humans, but it has been expanded for use in the veterinary field [81-89]. Thus far the use of PCR based detection technologies have been useful in monitoring treatment and vaccination strategies [86-88, 90], identification of symptomatic presentation [91, 92], and in the comparison of varying methods of Borrelia spp. detection including western blots, immunoflourescent assays, etc [82, 85, 88, 91-98]. The use of Borrelia spp. PCR based assays have not only impacted clinical and veterinary fields, but more diverse areas of inquiry into infectious organisms. Of note PCR has aided in the study of the epidemiologic compilation of data regarding the global distribution of various strains and virulence of Borrelia spp. [9, 14, 21, 26, 31, 39, 41, 42, 46, 47, 52, 62, 72-74, 83, 85, 98-110]. Furthermore, PCR continues to remain a vital lynch pin in the study of cell biologically relevant aspects of Borrelia spp. such as its antigenicity [111-114], presence in various vectors and detectability [44, 115], the expression of host and bacterial genes in response to infection [72, 105, 111, 112, 114, 116-125], mechanisms of transmission and infection [103, 106, 109, 110, 124, 126-129], and finally studies into cell biologically relevant differences in various Borrelia spp. strains and genospecies [110, 129-131]. Now PCR assays are finding their way into drug development and treatment monitoring that promises to usher in a new understanding in the ways to deal with Lyme disease world wide [132, 133]. Detection of Borrelia spp. by PCR is a fertile field for innovation and improvement with continual refinements, in both specific detection [2, 29, 54, 56, 134, 135] and techniques that aid in more accurate and reliable identification [10, 22, 28, 44, 46, 49, 54, 130, 134-137], being perpetually developed for use in both clinical and research laboratory settings. Due to this expansive body of growing knowledge much of the groundwork has already been laid for the use of Borrelia spp. PCR in a clinical diagnostic setting and we have developed our Borrelia spp. detection assay on this growing body of literature.

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