title: Amplification of ST50 gene using dry-reagent-based polymerase chain reaction for the detection of Salmonella typhi.
abstract: Conventional polymerase chain reaction (PCR) testing requires many pipetting steps and has to be transported and stored in cold chain. To overcome these limitations, we designed a ready-to-use PCR test for Salmonella typhi using PCR reagents, primers against the ST50 gene of S. typhi, a built-in internal amplification control (IAC), and gel loading dye mixed and freeze-dried in a single tube. The 2-step dry-reagent-based assay was used to amplify a 1238-bp target gene and an 810-bp IAC gene from 73 BACTEC blood culture broths (33 true positives for S. typhi and 40 true negatives for non-S. typhi). The sensitivity, specificity, positive predictive value, and negative predictive value of the PCR assay were 87.9%, 100%, 100%, and 90.9%, respectively. We suggest that this rapid 2-step PCR test could be used for the rapid diagnosis of typhoid fever.
authors.affiliation: Institute for Research in Molecular Medicine, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
authors.forename: Ismail
authors.initials: I
authors.lastname: Aziah
authors.forename: Manickam
authors.initials: M
authors.lastname: Ravichandran
authors.forename: Asma
authors.initials: A
authors.lastname: Ismail
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001769:Blood / Q000382:microbiology; D003470:Culture Media; D004587:Electrophoresis, Agar Gel; D005612:Freeze Drying / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents; D016133:Polymerase Chain Reaction / Q000379:methods*; D012485:Salmonella typhi / Q000235:genetics / Q000302:isolation & purification*; D013997:Time Factors; D014435:Typhoid Fever / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003470:Culture Media; D007202:Indicators and Reagents