Real-Time PCR Assay for Differentiation of Typhoidal and Nontyphoidal Salmonella

Rapid and accurate differentiation of Salmonella spp. causing enteric fever from nontyphoidal Salmonella is essential for clinical management of cases, laboratory risk management, and implementation of public health measures. Current methods used for confirmation of identification, including biochemistry and serotyping as well as whole-genome sequencing analyses, take several days. ABSTRACT Rapid and accurate differentiation of Salmonella spp. causing enteric fever from nontyphoidal Salmonella is essential for clinical management of cases, laboratory risk management, and implementation of public health measures. Current methods used for confirmation of identification, including biochemistry and serotyping as well as whole-genome sequencing analyses, take several days. Here we report the development and evaluation of a real-time PCR assay that can be performed directly on crude DNA extracts from bacterial colonies for the rapid identification of typhoidal and nontyphoidal Salmonella.

[1]  A. Zaidi,et al.  A Global Agenda for Typhoid Control—A Perspective from the Bill & Melinda Gates Foundation , 2019, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[2]  Sarah Lindsay,et al.  Evidence to Action: The 10th International Conference on Typhoid and Other Invasive Salmonelloses , 2019, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[3]  S. Baker,et al.  Evaluating PCR-Based Detection of Salmonella Typhi and Paratyphi A in the Environment as an Enteric Fever Surveillance Tool , 2018, The American journal of tropical medicine and hygiene.

[4]  S. Nair,et al.  First Report of CTX-M-15 Salmonella Typhi From England. , 2018, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[5]  S. Nair,et al.  Emergence of an Extensively Drug-Resistant Salmonella enterica Serovar Typhi Clone Harboring a Promiscuous Plasmid Encoding Resistance to Fluoroquinolones and Third-Generation Cephalosporins , 2018, mBio.

[6]  S. Nair,et al.  Characterization of new Salmonella serovars by whole-genome sequencing and traditional typing techniques. , 2016, Journal of medical microbiology.

[7]  Gemma C. Langridge,et al.  What’s in a Name? Species-Wide Whole-Genome Sequencing Resolves Invasive and Noninvasive Lineages of Salmonella enterica Serotype Paratyphi B , 2016, mBio.

[8]  Claire Jenkins,et al.  Identification of Salmonella for public health surveillance using whole genome sequencing , 2016, PeerJ.

[9]  M. Levine,et al.  Detection of Typhoidal and Paratyphoidal Salmonella in Blood by Real-time Polymerase Chain Reaction. , 2015, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[10]  Christopher M. Parry,et al.  Antimicrobial Resistance , and Antimicrobial Management of Invasive Salmonella Infections , 2015 .

[11]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[12]  J. Farrar,et al.  The sensitivity of real-time PCR amplification targeting invasive Salmonella serovars in biological specimens , 2010, BMC infectious diseases.

[13]  K. Hopkins,et al.  Rapid identification of Salmonella enterica subsp. arizonae and S. enterica subsp. diarizonae by real-time polymerase chain reaction. , 2009, Diagnostic microbiology and infectious disease.

[14]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[15]  R. Johnston,et al.  Salmonella paratyphi C: Genetic Divergence from Salmonella choleraesuis and Pathogenic Convergence with Salmonella typhi , 2009, PloS one.

[16]  Rosanna Lagos,et al.  PCR Method To Identify Salmonella enterica Serovars Typhi, Paratyphi A, and Paratyphi B among Salmonella Isolates from the Blood of Patients with Clinical Enteric Fever , 2008, Journal of Clinical Microbiology.

[17]  J. Wain,et al.  Detection of Vi-Negative Salmonella enterica Serovar Typhi in the Peripheral Blood of Patients with Typhoid Fever in the Faisalabad Region of Pakistan , 2005, Journal of Clinical Microbiology.

[18]  S. Nair,et al.  Salmonella enterica Serovar Typhi Strains from Which SPI7, a 134-Kilobase Island with Genes for Vi Exopolysaccharide and Other Functions, Has Been Deleted , 2004, Journal of bacteriology.

[19]  T. Ezaki,et al.  Selective Amplification of tyv (rfbE), prt (rfbS), viaB, and fliC Genes by Multiplex PCR for Identification of Salmonella enterica Serovars Typhi and Paratyphi A , 2002, Journal of Clinical Microbiology.

[20]  S. Martin Estimating disease prevalence and the interpretation of screening , 1984 .

[21]  A. B. CiusnE,et al.  Typhoid fever. , 1967, The Journal of the Arkansas Medical Society.