The polymerase chain reaction for the detection of borrelia burgdorferi in human body fluids

Abstract

Objective. To analyze clinical fluids for the presence of Borreliu butgdo#eri DNA using the polymerase chain reaction (PCR).Methods. We utilized a modified, nested PCR to detect the presence of Borreliu DNA in 99 samples of serum, urine, cerebrospinal fluid (CSF), or synovial fluid obtained from 44 patients with various stages of Lyme disease and 47 control subjects. Primer specificity was corroborated by examining 2 DNA data banks, testing against DNA from other organisms, and confirming results with a second set of nested primers.Results. Nested PCR was capable of detecting DNA from fewer than 10 organisms in 1 ml of fluid. The specificity of this technique was 96.4%, with a sensitivity of 76.7%. Although the specificity was uniformly high, the sensitivity was dependent upon the body fluid being tested: CSF loo%, urine loo%, synovial fluid 80%, and serum 59%. The rate of false-positive results was 3.6%.

Profile of patients with Baggio-Yoshinari Syndrome admitted at "Instituto de Infectologia Emilio Ribas"

“…The primers employed were targeted to identify outer surface protein A (OspA) 22 and genes for major flagellin FlaB and minor flagellin FlaA 20 , as well as for ribosomal 16S rRNA 3 .…”

Methodological Aspects

“…Glycogen (84,123) has been used successfully, and the sensitivity of PCR could be increased by the addition of 10 3 Leptospira biflexa organisms (7) to CSF before centrifugation. In CSF, the addition of hyaluronidase can further increase the sensitivity of the PCR (96). Although the DNA prepared by precipitation is of high purity, these methods are very time-consuming and labor intensive and are difficult to use routinely.…”

“…Although B. burgdorferi DNA could be detected in human body fluids from patients with late disease even 10 years or more after infection (22,75,84,93,96,114,123,155), the inability of PCR to confirm late disease has also been reported (38,68,107,128,195). Therefore, another way to define sensitivity might be to establish an expanded gold standard.…”

No detection of Borrelia burgdorferi-specific DNA in erythema migrans lesions after minocycline treatment

“…PCR is able to detect B. burgdorferi-specific DNA in a variety of clinical specimens from patients with both early and chronic manifestations of LB. Most reports about the application of PCR to urine specimens from patients with LB refer to patients with neuroborreliosis or late disease (3,7,10) and Lyme arthritis (3,12,30). PCR has already been performed on serum (5,12) and skin lesions from patients with EM (11, 16-18, 23, 27, 29), but only a few, preliminary observations exist about investigations of urine (11,25).…”

“…Most reports about the application of PCR to urine specimens from patients with LB refer to patients with neuroborreliosis or late disease (3,7,10) and Lyme arthritis (3,12,30). PCR has already been performed on serum (5,12) and skin lesions from patients with EM (11, 16-18, 23, 27, 29), but only a few, preliminary observations exist about investigations of urine (11,25). We report here on 26 consecutively diagnosed cases of EM in patients residing in an area where borrelial infections are endemic (Styria, Austria).…”