PCRmax Plasmodium (all species) Instruction Manual

18S ribosomal RNA (18S) gene

Plasmodium (all
species)

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Plasmodium is a genus of obligate, protozoan parasites that cause Malaria. There are over
200 known species within this genus with four infecting humans (P.falciparum, P.vivax, P.
malariae, P.ovale) and others targeting a variety of vertebrate hosts as well as exploiting a
number of mosquito species during their life cycle.

Mosquitoes of the Aedes, Culex, Mansonia, Theobaldia and Anopheles genera transmit
the parasite to a vertebrate host by biting. The sporozoites of the parasite are injected into
the individual with the mosquito saliva. These sporozoites target the host liver where they
replicate asexually within 24 hours, 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 macro­gametes 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
gland ready to infect another individual when the mosquito feeds again.

Symptoms of human plasmodium infections include fever, chills, weakness, headache,
vomiting, diarrhoea, anaemia, and pulmonary and renal dysfunction. Treatment is based
on identification of the specific species causing the infection, where in the world the
infection was acquired and drug resistance but if left untreated infection may cause death.

Introduction to Plasmodium (all species)

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MIN

MAX

Our kit for Plasmodium species has been designed for the specific and exclusive in vitro
quantification of all plasmodium species within the subgensus Plasmodium
(Plasmodium). The 18S ribosomal gene, is the ideal target to achieve a broad based
detection profile for all species of plasmodium within this subgenus. 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 (all species) (Plasmodium_spp) genomes is
designed for the in vitro quantification of Plasmodium_spp genomes. The kit is designed
to have the broadest detection profile possible whilst remaining specific to the
Plasmodium_spp genome.

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

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

FAM labelled

• Plasmodium_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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