PCRmax Plasmodium falciparum Instruction Manual

plasmepsin 4 gene

Plasmodium
falciparum

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Plasmodium is a genus of protozoa that cause Malaria. Plasmodium species use two hosts
in their lifecycle. Plasmodium falciparum, which infects humans, uses female Anophele
mosquitoes as a vector. The parasite has a double-stranded DNA genome spread over 14
chromosomes.

As the mosquito bites a human, the sporozoites of the parasite are injected into the
individual with the mosquito saliva. The sporozoites target the host liver where they
replicate asexually creating merozoites which are subsequently released into the host
circulatory system where they invade erythrocytes and undergo nuclear division causing
these cells to burst. The parasite can also reproduce sexually producing gametocytes
which are then able to infect the vector host when they bite the human. In the mid gut of
the mosquito these mature into micro-gametes with flagella and macro-gametes that fuse
together to produce a zygote which later develops into an oocyst in the space between the
epithelial cells and the basal lamina. These oocysts undergo asexual replication to produce
sporozoites of around 1µm in diameter which relocate to the salivary glands ready to infect
another individual when the mosquito feeds again.

After an incubation period varying between 7 and 30 days, infection with P. falciparum
presents initially with fever, muscle pain, nausea. Symptoms including anaemia and
confusion can also occur. Treatment is based on identification of the specific species
causing the infection, where in the world the infection was acquired and drug resistance. P.
falciparum infections are most commonly treated with Chloroquin.

Introduction to Plasmodium falciparum

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MIN

MAX

The target sequence within the plasmepsin 4 gene has very low homology with other
species of plasmodia that cause malaria. 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.

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

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

• P.falciparum specific primer/probe mix (150 reactions BROWN)

FAM labelled

• P.falciparum 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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