Development and evaluation of a broad-range PCR-ELISA assay with Borrelia burgdorferi and Streptococcus pneumoniae as model organisms for reactive arthritis and bacterial meningitis

Citation
C. Fischer-romero et al., Development and evaluation of a broad-range PCR-ELISA assay with Borrelia burgdorferi and Streptococcus pneumoniae as model organisms for reactive arthritis and bacterial meningitis, J MICROB M, 40(1), 2000, pp. 79-88
Citations number
19
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,Microbiology
Journal title
JOURNAL OF MICROBIOLOGICAL METHODS
ISSN journal
0167-7012 → ACNP
Volume
40
Issue
1
Year of publication
2000
Pages
79 - 88
Database
ISI
SICI code
0167-7012(200003)40:1<79:DAEOAB>2.0.ZU;2-R
Abstract
We have developed an assay based on a 16S rDNA broad-range amplification sy stem followed by species-specific detection with a commercially available P CR-ELISA kit. B. burgdorferi and S. pneumoniae were used as model systems f or arthritis and meningitis, respectively. The sensitivity of the B. burgdo rferi assay was comparable to that of a species-specific PCR, whereas for S . pneumoniae the detection limit was one to three organisms as determined b y plate counts. To specifically differentiate two species, two discontinuou sly located nucleotide differences in the region complementary to the captu re probe are required during the detection step with the PCR-ELISA lilt. A preliminary clinical evaluation was performed with eight specimens (joint a nd cerebrospinal fluids) previously shown to contain B. burgdorferi DNA. Ex cept for one sample which was positive by the broad-range PCR-ELISA system only, the results were in agreement with those obtained by B. burgdorferi s pecies-specific PCR. None of the 23 control samples were positive by either method. Thus, broad-range amplification in combination with the PCR-ELISA kit promises to be a sensitive and specific format for the detection of age nts causing reactive arthritis, meningitis or other diseases associated wit h a limited number of different bacteria. (C) 2000 Elsevier Science B.V. Al l rights reserved.