PCRmax Entamoeba histolytica Instruction Manual

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Entamoeba histolytica

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Entamoeba histolytica is an anaerobic, protozoan, intestinal parasite responsible for a
disease called amoebiasis. It usually occurs in the large intestine and causes internal
inflammation. It belongs to the genus Entamoeba and class Archamoeba. Amongst
parasitic diseases, E. histolytica is one of the leading causes of morbidity and mortality in
developing countries. E. histolytica is transmitted by ingestion of exit body containing cysts
from faecally contaminated food and water or from hands. Due to their protective walls, the
cysts can remain viable for several weeks in external environments. Species within this
genus are small, single celled organisms with an anterior bulge representing a lobose
pseudopod. The E. histolytica trophozoites are oblong and approximately 15-20µM in
length, whereas the cysts are spherical and typically 12-15 µM in diameter.

Entamoeba cysts are most commonly transmitted by ingestion so must be extremely
robust to survive the hostile environment of the stomach. The cysts transform to
trophozoites in the small intestine where they multiply by binary fission to then colonise the
large intestine. They cause major calcium ion influx to the cells of the large intestine
resulting in cell death and ulcer formation. The Trophozoites subsequently form new cysts
which are excreted once more in faeces.

Infection with E. histolytica generally causes mild symptoms such as abdominal pain,
flatulence and diarrhoea, but more severe infections can lead to amoebosis. This is a
condition encompassing amoebic dysentery characterized by severe abdominal pain, fever
and blood in the faeces and less commonly amoebic liver abscesses.

Introduction to Entamoeba histolytica

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MIN

MAX

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

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

• E.histolytica specific primer/probe mix (150 reactions BROWN)

FAM labelled

• E.histolytica 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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