A highly sensitive and fast non-radioactive method for the detection of polymerase chain reaction products from Salmonella serovars, such as Salmonella typhi, in blood specimens

Abstract

A polymerase chain reaction based test was developed for the detection of Salmonella spp. in blood specimens. After amplification of a 389 bp-polymerase chain reaction product from the invA gene, a microtiter plate hybridization assay was performed. The protocol described allowed the detection of six to seven copies of the Salmonella typhi genome, as determined by serial dilutions of DNA from S. typhi. Eighteen blood specimens from artificially infected rats and 22 blood specimens from patients were analyzed to validate the method. Considering that the most frequent Salmonella serovar isolated from blood in case of bacteremia is S. typhi, the polymerase chain reaction-microtiter plate hybridization technique could be used as a novel, rapid diagnostic method for typhoid fever, particularly when standard culture assays are negative.