PCRmax Salmonella enterica Instruction Manual

Invasine A gene (InvA)

Salmonella enterica

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Salmonella enterica is a motile, rod shaped, Gram-negative bacterium of the
Enterobecteriacae family. There are over 2000 serovars that make up this species with
genomes ranging in length from 4.5-5 M nucleotides arranged in a circular configuration
with plasmids. These serovars can cause illnesses in human ranging from gastroenteritis
to typhoid fever which kills around 500,000 people worldwide per year.

S. enterica can be found naturally on raw poultry and red meat and can contaminate some
water sources. When ingested from any of these sources, the bacterium secretes adhesion
proteins called adhesins that mediate attachment to the host intestinal cells. Once within
the host cells, the bacteria translocate to the liver and spleen where bacterial replication
continues. From the liver, bacteria are release back into the intestine and are subsequently
excreted. In areas with poor hygiene and water treatment, the bacteria can infect water
sources at this time. A number of antibiotics have proven useful in reducing the length of
infection.

The 3 main serovars cause widely different symptoms. Salmonella enterica serovar Typhi
causes typhoid fever which initially presents with vomiting and diarrhea. This disease
progresses over several weeks leading to a high fever, abdominal pain and a rash on the
chest or abdomen. Complications arise from intestinal hemorrhaging or perforation and
inflammation of organs and bones.
Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis both
cause an infection with much milder symptoms than S. typhi, and are responsible for
causing salmonellosis, a type of gastroenteritits, with S. enteritidis being much more
common than S. typhimurium. This disease is characterised by vomiting, diarrhea and
abdominal pain but usually only last around 7 days before the immune system fights off the
infection. This is not the case in people with weak immune systems such as the elderly or
infants in whom the disease may become fatal if not treated.

Introduction to Salmonella enterica

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MIN

MAX

The Salmonella enterica primers have been designed for the specific and exclusive in
vitro quantification of all Salmonella enteric subsp. enterica strains in which invasin A has
been described. The target sequence (invA) has previously been shown to be a good
genetic marker for broad detection of pathogenic salmonella strains in other real time
PCR based studies (Hadjinicolaou AV, et.al 2009). The primers and probe sequences in
this kit have 100% homology with a broad range of clinically relevant reference
sequences based on a comprehensive bioinformatics analysis.

The PCR Max qPCR Kit for Salmonella enterica (S.enterica) genomes is designed for the
in vitro quantification of S.enterica genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the S.enterica genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
S.enterica sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• S.enterica specific primer/probe mix (150 reactions BROWN)

FAM labelled

• S.enterica positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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