PCRmax Salmonella (all pathogenic species) Instruction Manual

invasion protein (invA) gene

All pathogenic
Salmonella species

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Salmonella spp. are members of the family Enterobacteriaceae. They are Gram-negative,
facultatively anaerobic, flagellated, rod-shaped organisms. They are approximately 0.7 to

1.5 µm in diameter and 2 to 5 µm in length and responsible for a large number of cases of
foodborne illness throughout the world. Salmonella have circular DNA genomes with a
mean length of approximately 4530 kb, although this can vary by up 1000 kb. Salmonella
classification is extremely complex, however, the genus is divided into two species: S.
enterica and S.bongori. S. enterica is then itself divided into 6 biochemically distinct
subspecies and the Salmonella genus is further classified into serovars (serotypes) based
on the lipopolysaccharide (O), flagella protein (H), and sometimes the capsular (VI)
antigens. There are more than 2500 known serovars and within a serovar there may be
strains that differ in virulence.

Salmonella are mainly transmitted by the faecal-oral route. They are carried
asymptomatically in the intestines or gall bladder of many animals, being continuously or
intermittently shed in the faeces. Humans can become infected if they do not wash their
hands after contact with infected animals or animal faeces. In such instances the bacteria
adhere to and enter the cells of the intestinal epithelium. The toxins produced by the
bacteria can damage and kill the cells that line the intestines, which results in intestinal
fluid loss. The bacteria can survive for weeks in a dry environment and far longer in water
thus they are frequently present in polluted waters. Salmonella can also be carried latently
in the mesenteric lymph nodes or tonsils; these bacteria are not shed, but can become
reactivated after stress or immunosuppression. In addition, fomites and vectors can spread
Salmonella and vertical transmission occurs in birds, with contamination of the vitalize
membrane, albumen and possibly the yolk of eggs. Salmonella spp. can also be
transmitted in utero in mammals.

There are two different disease conditions that are distinct to salmonellosis; gastroenteritis
and enteric typhoid fever. The gastroenteritis is a nonsystemic infection of the intestinal
tract and regional lymph nodes that gives rise to headache, muscle aches, diarrhoea,
vomiting, abdominal cramping, chills, fever, nausea and dehydration. In contrast, the
enteric typhoid fever is a systemic disease in which the microorganism replicates within the
cells of the reticuloendothelial system. The symptoms usually appear 6 to 72 hours after
ingesting contaminated food although individuals can be infected with the bacteria without
having symptoms. Those with and without symptoms shed the bacteria in their stool and it
is important that personal hygiene be maintained at all times.

Introduction to All pathogenic Salmonella species

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MIN

MAX

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

The primers and probe sequences in this kit have 100% homology with a broad range of
Salmonella_invA 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

• Salmonella_invA specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Salmonella_invA 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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