Comparison of primers for the detection of Salmonella enterica serovars using real-time PCR

Letters in Applied Microbiology
A T CsordasM J Delwiche

Abstract

To evaluate the specificity and sensitivity of PCR primers for the detection of Salmonella enterica in a real-time PCR assay using pure cultures. Unenriched whole cells in sterile water were used as template for each PCR. SYBR Green dye was used for the nonspecific detection of dsDNA. The real-time PCR detection limits of five previously published primer sets used in conventional PCR applications were not below 3 x 10(3) CFU per reaction (rxn). A new primer set, Sen, was designed, which detected Salm. enterica Newport down to 6 CFU rxn(-1) in one case, and gave an average detection limit of 35 CFU rxn(-1) over three separate runs. Primers originally designed for end-point PCR did not have adequate specificity or sensitivity compared with those specifically designed for real-time PCR. This study emphasizes the importance of evaluating real-time PCR primer sets in pure cultures prior to testing in field samples. This study will benefit other researchers in selecting an appropriate primer set for real-time PCR detection of Salm. enterica.

References

Apr 1, 1992·Bio/technology·R HiguchiR Griffith
May 1, 1997·Journal of Clinical Microbiology·A J BäumlerR Reissbrodt
Feb 5, 1998·Journal of Applied Microbiology·R F WangC E Cerniglia
Sep 9, 1999·The Journal of Infectious Diseases·J C Mohle-BoetaniD J Vugia
Oct 6, 1999·Molecular and Cellular Probes·C M Gooding, P V Choudary
Sep 2, 2000·Journal of Clinical Microbiology·J HoorfarP Rådström
Aug 21, 2001·Annals of Internal Medicine·J C Mohle-BoetaniUNKNOWN Investigation Team
Sep 27, 2002·Applied and Environmental Microbiology·J D BarakA O Charkowski
Jan 7, 2003·Applied and Environmental Microbiology·Burkhard MalornyReiner Helmuth
Oct 24, 2003·Journal of Food Protection·Jeri D BarakDaniel C Mills
Oct 29, 2003·BioTechniques·Weijie LiUlrich K Schubart
Nov 25, 2003·Letters in Applied Microbiology·C J Ziemer, S R Steadham

❮ Previous
Next ❯

Citations

Jul 22, 2005·Letters in Applied Microbiology·J D BarakM J Delwiche
May 22, 2009·Acta Veterinaria Hungarica·Ching-Yang ChengChin-Cheng Huang
Jun 30, 2010·Expert Review of Medical Devices·Louise O'Connor, Barry Glynn
May 5, 2007·Journal of Food Protection·Benjamin R WarrenKeith R Schneider

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.