PCRmax Acanthamoeba_spp Instruction Manual

18S ribosomal RNA (18S) gene

Acanthamoeba_spp

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Acanthamoeba is a genus of amoeba containing both free-living and parasitic species. The
free-living species are bacterivores gaining nutrients and energy from bacterial
consumption and the parasitic species infect animals, sometimes causing disease.
Species of this genus are single-celled organisms ranging in size from 15 to 35um in
length, with a flexible cell wall allowing for shape changes from round or oval to triangular.
There are two stages to the Acanthamoeba life cycle: cysts and trophozoites which are
infectious to animals and can cause cutaneous infections, encephalitis or keratitis.

Breaks in the skin provide Acanthamoeba species access to the host body where they can
cause a localised infection or spread via the blood to organs or the central nervous system.
Infection triggers an immune response which can cause neuronal damage which may be
fatal. Cysts can also enter the body by inhalation and once within the airways transforms to
the next stage of its life cycle. The cyst wall disappears, the nucleus divides twice and the
protozoan enters the infectious, trophozoite stage. Trophozoites replicate by mitosis and
later develop into a new cyst with a glycogen vacuole and outer wall which is capable of
infecting new hosts.

Infections of the skin or organs are generally seen in individuals with weakened immune
systems and so are rare. Granulomatus Encephalitis can result in symptoms including
headaches, fever, seizures and altered mental state leading to coma and death within
several months of infection.

Infection of the eye is seen in individuals with fully functioning immune systems and most
commonly in individuals who wear contact lenses. The infection results in keratitis which
can cause permanent blindness. Symptoms of this infection include eyes pain, blurred
vision and sensitivity to light.

Antimicrobial treatment can only offer limited success in treatment of infection due to the
toughness of the cyst.

Introduction to Acanthamoeba_spp

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MIN

MAX

Our kit for Acanthamoeba_spp has been designed for the specific and exclusive in vitro
quantification of this genus . The 18S ribosomal gene, is the ideal target to achieve a
broad based detection profile for all species within this genus. 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. Other closely
related species are not detected.

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

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

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

FAM labelled

• Acanthamoeba_spp 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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