PCR in laboratory diagnosis of human Borrelia burgdorferi infections

Abstract

The laboratory diagnosis of Lyme borreliosis, the most prevalent vector-borne disease in the United States and endemic in parts of Europe and Asia, is currently based on serology with known limitations. Direct demonstration of Borrelia burgdorferi by culture may require weeks, while enzyme-linked immunosorbent assays for antigen detection often lack sensitivity. The development of the PCR has offered a new dimension in the diagnosis. Capable of amplifying minute amounts of DNA into billions of copies in just a few hours, PCR facilitates the sensitive and specific detection of DNA or RNA of pathogenic organisms. This review is restricted to applications of PCR methods in the diagnosis of human B. burgdorferi infections. In the first section, methodological aspects, e.g., sample preparation, target selection, primers and PCR methods, and detection and control of inhibition and contamination, are highlighted. In the second part, emphasis is placed on diagnostic aspects, where PCR results in patients with dermatological, neurological, joint, and ocular manifestations of the disease are discussed. Here, special attention is given to monitoring treatment efficacy by PCR tests. Last, specific guidelines on how to interpret PCR results, together with the advantages and limitations of these new techniques, are presented.

Detection ofBorrelia burgdorferi sensu stricto by reverse line blot in the joints of Dutch patients with Lyme arthritis

“…However, this is unlikely, since all synovial samples spiked with B burgdorferi sensu lato DNA indicated the absence of such PCR inhibitors. We cannot completely exclude the possibility that the method of DNA extraction or the low frequency of the genetic target chosen in this study may have resulted in a false-negative result, although the sensitivity of the PCR with RLB used in this study was comparable with that found in most other studies (44).…”

“…Another cause that should be considered as an explanation for the negative PCR result in the 1 patient with Lyme arthritis is interference by host DNA in the Lyme-specific PCR (44). However, this is unlikely, since all synovial samples spiked with B burgdorferi sensu lato DNA indicated the absence of such PCR inhibitors.…”

The Role of Culture and Nucleic Acid Amplification in Diagnosis of Lyme Borreliosis

“…Since then, a variety of PCR assays aimed to detect Lyme borrelia DNA in clinical specimens have been developed and evaluated for their diagnostic value in patients with different clinical manifestations of Lyme borreliosis. Many technical details and applications of PCR assays were addressed in several recently published reviews (Schmidt, 1997;Bunikis and Barbour, 2002;Wang, 2002;Dumler, 2003;Wilske, 2003;Aguero-Rosenfeld et al, 2005).…”

“…A variety of clinical specimens from patients with suspected Lyme borreliosis have been analyzed by PCR assays (Schmidt, 1997;Aguero-Rosenfeld et al, 2005). Of these, skin biopsy samples taken from patients with EM were the most frequently tested specimens Wienecke et al, 1993;Muellegger et al, 1995Muellegger et al, , 1996von Stedingk et al, 1995;Picken et al, 1997;Rijpkema et al, 1997;Brettschneider et al, 1998;Oksi et al, 2001;Liveris et al, 2002;Wang, 2002).…”

“…Although many Lyme borrelia genes have been amplified and analyzed by PCR in research laboratories, only a few of these target genes have been utilized for detection of Lyme borrelia DNA in clinical specimens. Among these are the chromosomally encoded ribosomal DNA, flaB, recA, and p66 and the plasmidencoded ospA (Schmidt, 1997).…”