PCRmax Plasmodium knowlesi Instruction Manual

plasmepsin gene

Plasmodium knowlesi

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 knowlesi, which infects humans, uses Anophele mosquitoes
as a vector. This species is most commonly found in South-East Asia.

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 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.

After an incubation period of around 10 to 12 days, infection with P. knowlesi 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.

Introduction to Plasmodium knowlesi

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MIN

MAX

The target sequence within the plasmepsin 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 knowlesi (P.knowlesi ) genomes is designed for
the in vitro quantification of P.knowlesi genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the P.knowlesi genome.

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

FAM labelled

• P.knowlesi 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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