title: Rapid detection of Salmonella Typhi by loop-mediated isothermal amplification (LAMP) method.
abstract: An in-house loop-mediated isothermal amplification (LAMP) reaction was established and evaluated for sensitivity and specificity in detecting the presence of Salmonella Typhi (S. Typhi) isolates from Kelantan, Malaysia. Three sets of primers consisting of two outer and 4 inner were designed based on locus STBHUCCB_38510 of chaperone PapD of S. Typhi genes. The reaction was optimised using genomic DNA of S. Typhi ATCC7251 as the template. The products were visualised directly by colour changes of the reaction. Positive results were indicated by green fluorescence and negative by orange colour. The test was further evaluated for specificity, sensitivity and application on field samples. The results were compared with those obtained by gold standard culture method and Polymerase Chain Reaction (PCR). This method was highly specific and -10 times more sensitive in detecting S. Typhi compared to the optimised conventional polymerase chain reaction (PCR) method.
authors.affiliation: Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
authors.forename: J
authors.initials: J
authors.lastname: Abdullah
authors.affiliation: Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
authors.forename: N
authors.initials: N
authors.lastname: Saffie
authors.affiliation: Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
authors.forename: F A R
authors.initials: FA
authors.lastname: Sjasri
authors.affiliation: Hospital Raja Perempuan Zainab II 15586 Kota BharuKelantan Malaysia Hospital Raja Perempuan Zainab II, 15586 Kota Bharu, Kelantan, Malaysia.
authors.forename: A
authors.initials: A
authors.lastname: Husin
authors.affiliation: School of Medical Sciences Universiti Sains Malaysia Kubang Kerian Kelantan School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan.
authors.forename: Z
authors.initials: Z
authors.lastname: Abdul-Rahman
authors.affiliation: Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
authors.forename: A
authors.initials: A
authors.lastname: Ismail
authors.affiliation: Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
authors.forename: I
authors.initials: I
authors.lastname: Aziah
authors.affiliation: Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
authors.forename: M
authors.initials: M
authors.lastname: Mohamed
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D008296:Malaysia; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012485:Salmonella typhi / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014435:Typhoid Fever / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D017931:DNA Primers
keywords: Salmonella Typhi; loop-mediated isothermal amplification (LAMP)