PCRmax Aeromonas hydrophila Instruction Manual

aerolysin (aerA)

Bacteria Hemorrhagic
Septicemia

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Aeromonas hydrophila is a heterotrophic, gram-negative bacterium of the
Aeromonadaceae family. This bacterium has a rod shape with rounded ends reaching 1

-3μm in length. Polar flagella allow motility. 3 enterotoxins encoded by the 5Mbp genome
are thought to be responsible for making this bacterium pathogenic. These entereotoxins
are namely: cytotoxic (act); heat-labile cytotonic (alt) and heat-stable cytotonic (ast) of
which the former is thought to contribute most to toxicity. A. hydrophila can be considered
as an opportunistic pathogen which is linked to infections in fish and amphibians as well as
humans, but may not be the cause.

Infection most usually occurs when an organism is dealing with environmental change.
This could be a change in temperature or when infected with a different bacterium. In fish,
the bacteria mainly infect the kidneys and liver resulting in disease that is thought to be the
result of stress. In amphibians, the infection can cause internal heamorrhaging. Infection in
humans can be caused by ingestion of infected food such as fish or meat. Once ingested
the bacterium progresses to an organ via the bloodstream. Alternatively the bacterium can
infect open wounds. The bacteria have pili which they utilise in attachment to host
organism and invasion of the cells or organs. Colonisation occurs by asexual reproduction.
Upon cell invasion, the enterotoxins are activated causing cell damage.
Transmission in fish occurs when infected fish are moved into new fisheries without
sanitation.
The bacteria are largely antibiotic resistant, can survive refrigeration and are also able to
grow at temperatures in a wide range up to 37°C. However, a sodium/calcium hypochlorite
solution can kill this bacterium.

Infection in fish may cause diseases including tail and fin rot, heamorrahagic septicemia
and ulcer disease. In amphibians this bacterium is linked to a disease called red leg which
is a result of internal heamorrhaging. Infection in humans can lead to a number of diseases
including septicemia, meningitis, pneumonia and gastroenteritis. The bacteria can also
infect open wounds causing cellulitis, myonecrosis and ecthyma gangrenosum. The latter
two diseases occur more commonly in the immunocompromised, although infections do
also occur in immunocompetent people.

Introduction to Bacteria Hemorrhagic Septicemia

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MIN

MAX

The primers have 100% homology with >95% of sequenced Aeromonas hydrophila
isolates in the NCBI database. Due to the high sequence homology, the primers also
have a high level of homology to other closely related Aeromonas sp. The primers
therefore may also detect A. salmonicida, A. caviae and A. sobria.

The PCR Max qPCR Kit for Bacteria Hemorrhagic Septicemia (A.hydrophila) genomes is
designed for the in vitro quantification of A.hydrophila genomes. The kit is designed to
have the broadest detection profile possible whilst remaining specific to the A.hydrophila
genome.

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

• A.hydrophila specific primer/probe mix (150 reactions BROWN)

FAM labelled

• A.hydrophila 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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