Detection of Yersinia pestis using branched DNA.
In contrast to target amplification methods, e.g. polymerase chain reaction, the branched DNA (bDNA) signal amplification method quantitates target nucleic acid at physiological levels, involving a series of hybridization reactions without thermal cycling. In this report, we describe a modification of the bDNA assay in which a <<concatenated>> preamplifier oligonucleotide (206 mer) is used in concert with ELISA and light addressable potentiometric sensor (LAPS) formats to detect the plasminogen activator (pla) gene of Yersinia pestis, the etiological agent of plague. Pla is encoded by a 9.6-kb plasmid pPCP, which is essential for virulence. The detection limit of the bDNA-ELISA and LAPS assays is less than 10 000 and 1000 molecules of Y. pestis plasmid DNA, respectively.
Iqbal
S S
SS
Division of Life Sciences, University of Texas at San Antonio, San Antonio, TX, 78249, USA.
Chambers
J P
JP
Brubaker
R R
RR
Goode
M T
MT
Valdes
J J
JJ
D001426:Bacterial Proteins; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis; D008969:Molecular Sequence Data; D010960:Plasminogen Activators / Q000235:genetics; D015010:Yersinia pestis / Q000235:genetics / Q000302:isolation & purification*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C078140:Pla protease, Yersinia pestis; D010960:Plasminogen Activators