PCRmax Entamoeba (all species) Instruction Manual

18S ribosomal RNA (18S) gene

Entamoeba

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Entamoeba is a genus of parasitic protozoan of the Archamoeba class. Species within this
genus are small, single celled organisms with an anterior bulge representing a lobose
pseudopod. Most species can be found in the intestines of the animals that they infect,
except for E. gingivalis which lives in the mouth, and are commensal. However, E.
histolytica is known to cause amoebiasis.
Entamoeba cysts are inhaled or ingested in contaminated food sources and enters the
host through the digestive tract after surviving the hostile environment of the stomach.
Cysts of different species vary in the number of nuclei they contain and as such, the
number of trophozoite forms they produce after excystation in the small intestines. These
trophozites, 10-20m in diameter, multiply by binary fission and colonise the large
intestine causing major calcium ion influx to the cells here resulting in cell death and ulcer
formation. Trophozoites subsequently form new cysts which are excreted and remain
viable in the environment for up to several months.
Infection with E. histolytica generally causes mild symptoms such as abdominal pain and
diarrhoea but more severe infections can lead to amoebosis, a condition encompassing
amoebic dysentery characterized by severe abdominal pain, fever and blood in the feaces
and less commonly amoebic liver abscesses.

Introduction to Entamoeba

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MIN

MAX

Our kit for Entamoeba_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 Entamoeba (Entamoeba_spp) genomes is designed for the in
vitro quantification of Entamoeba_spp genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the Entamoeba_spp genome.

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

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

FAM labelled

• Entamoeba_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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