PCRmax Acanthamoeba castellanii Instruction Manual

Acanthamoeba
castellanii

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Acanthamoeba castellanii is a unicellular, opportunistic human parasite and belongs to a
genus of amoebae which frequently inhabit freshwater. It is one of the most common soil
protozoa.

The A.castellanii life cycle consists of 2 stages: a trophpozoite (14-40µm in diameter) and
a cyst which has a double layered wall of 12-16µm in diameter.
Acanthamoeba species have been classified into 3 morphologic groups based on the size
of their cysts. Group1 having the larger cysts and the cyst size decreasing through Groups
2 and 3. The major human pathogens belong to group 2. A.castellanii can lyse bacteria
and produce a vast range of enzymes including chitinases and cellulases.

Several species of Acanthamoeba including A.castellanii have been known to cause
amoebic keratitis. Studies suggest that the pathogenic potential of A.castellanii to cause
keratitis could be related to its ability to bind to corneal epithelium. As amoebic keratitis is a
potentially blinding infection of the cornea it is crucial that the disease is diagnosed and
treated as early as possible.

Introduction to Acanthamoeba castellanii

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MIN

MAX

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

The primers and probe sequences in this kit have 100% homology with a broad range of
A.castellanii 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.castellanii specific primer/probe mix (150 reactions BROWN)

FAM labelled

• A.castellanii 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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