Application of Random Amplified Polymorphic DNA analysis to differentiate strains ofSalmonella typhiand otherSalmonellaspecies

Abstract

A Random Amplified Polymorphic DNA (RAPD) fingerprinting method was developed to differentiate isolates of Salmonella serotype typhi (S. typhi) and other Salmonella isolates. A panel of five primers was used to examine 63 isolates of Salm. typhi, including 56 strains isolated in Taiwan and seven strains obtained abroad. Twenty‐one RAPD types were revealed using the RAPD fingerprinting method. An RAPD with primer 6032 yielded a polymorphism in a 350 bp fragment that differentiated the attenuated vaccine strain Salm. typhi Ty21a from the rest of the Salm. typhi strains. Strains of Salm. typhi were divided into five types with primer D14307. Primer D14307 also proved capable of discrimination among 65 other Salmonella isolates representing 42 different serotypes. The bacterial DNA used in this RAPD protocol was obtained using a commercially available DNA extraction kit (GeneReleaser). The DNA of various strains of Salmonella from this simple extraction procedure could be discriminated within a few hours using the RAPD technique.

Random Amplified Polymorphic DNA and Phenotyping Analysis of Salmonella enterica Serovar Enteritidis Isolates Collected from Humans and Poultry in Uruguay from 1995 to 2002

“…It offers good power for discrimination within several Salmonella serotypes, such as Dublin (20), Panama (46), Typhimurium (4), Typhi (43), and Enteritidis (7, 21,22,24,25,28). One appeal of this method is that a large number of arbitrarily selected primers can be tested to identify those that might be suitable for a particular application.…”

“…Since 1995, RAPD-PCR analysis (53,54) has been used either to differentiate Salmonella serotypes or to distinguish strains within a single serotype (3,5,12,15,18,31,43,47). It offers good power for discrimination within several Salmonella serotypes, such as Dublin (20), Panama (46), Typhimurium (4), Typhi (43), and Enteritidis (7, 21,22,24,25,28).…”

Serotyping and molecular typing of Salmonella species isolated from wastewater in Nsukka, Nigeria

“…The use of DNA-based typing methods is becoming increasingly important in epidemiological survey and differentiation of Salmonella species. Some of these methods include pulsed field gel electrophoresis (PFGE) (Mohand et al, 1999;Kubota et al, 2005), PCR ribotyping (Lagatolla et al, 1996), automated nuclease PCR assay (Hoorfar et al, 2000) and random amplification of polymorphic DNA (RAPD) (Shangkuan and Lin, 1998;Smith et al, 2011). RAPD-PCR does not require any specific knowledge of the target DNA sequence, making it a flexible and powerful tool with general applicability.…”

Development of Microarray and Multiplex Polymerase Chain Reaction Assays for Identification of Serovars and Virulence Genes in Salmonella Enterica of Human or Animal Origin

“…40 To identify the individual serovars (based on O and H antibody-antigen tests) of Salmonella, testing is routinely performed in state reference laboratories and in several veterinary laboratories, which, because of the high number of samples submitted, can lead to long turnaround times. In an attempt to streamline the process, molecular techniques have been developed, such as ribotyping, 14 polymerase chain reaction (PCR) single-strand conformation polymorphism analysis, 34 restriction fragment length polymorphism, 23 pulsed-field gel electrophoresis, 4,25 IS200 fingerprinting, 15 automated 59 nuclease PCR, 24 and random amplified polymorphic DNA analysis, 43 which are major improvements in serovar determination, but the success of these assays requires highly technical skills, are prone to interlaboratory variation, are time consuming, and are incapable of processing large numbers of samples. Multiplex PCR assays are efficient platforms for detecting many gene targets in a single-sample preparation by incorporating multiple primer pairs.…”