RT Journal Article
T1 Development and evaluation of a PCR-enzyme-linked immunosorbent assay for diagnosis of human brucellosis.
A1 Morata, Pilar
A1 Queipo-Ortuño, María I.
A1 Reguera, Jose M.
A1 García-Ordoñez, Miguel A.
A1 Cárdenas, Ana
A1 Colmenero, Juan D
K1 Brucelosis
K1 Enzimoinmunoanálisis por adsorción
K1 Humanos
K1 Reacción en cadena de la polimerasa
K1 Juego de reactivos para diagnóstico
K1 Reproducibilidad de los resultados
K1 Sensibilidad y especificidad
AB In order to overcome some of the limitations of conventional microbiological techniques in the diagnosis of human brucellosis, a simple PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed. After amplification of a 223-bp sequence of a gene that codes for the synthesis of an immunogenetic membrane protein specific for the Brucella genus (BCSP31), the digoxigenin-labeled amplified product was hybridized with a biotinylated capture probe which was complementary to the inner part of the amplicon. The hybrid was captured on streptavidin-coated microtiter plates and detected by using an antidigoxigenin Fab-peroxidase conjugate. The detection limit of the PCR-ELISA in a background of 3.5 micro g of human genomic DNA was 10 fg (two bacterial cells). The PCR-ELISA showed an analytical sensitivity higher than that of ethidium bromide staining and equal to that obtained by conventional PCR followed by dot blot hybridization. In 59 peripheral blood samples from 57 consecutive patients with active brucellosis and 113 control samples, the PCR-ELISA was found to be 94.9% sensitive and 96.5% specific, whereas the sensitivity of the blood culture was only 70.1%. Since the assay can be performed in 1 day, is very reproducible, is easily standardized, and avoids the risk of infection in laboratory workers, this PCR-ELISA seems to be a practical and reliable tool for the diagnosis of human brucellosis.
PB American Society for Microbiology
SN 0095-1137
YR 2003
FD 2003-01
LK http://hdl.handle.net/10668/2450
UL http://hdl.handle.net/10668/2450
LA en
NO Morata P, Queipo-Ortuño MI, Reguera JM, García-Ordoñez MA, Cárdenas A, Colmenero JD. Development and evaluation of a PCR-enzyme-linked immunosorbent assay for diagnosis of human brucellosis. J Clin Microbiol. 2003; 41(1):144-8
NO Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't;
DS RISalud
RD Apr 10, 2025