PCRmax Plasmodium vivax Instruction Manual

aspartic protease PM4

Plasmodium vivax

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Plasmodium is a genus of protozoa that cause Malaria. Plasmodium species use two hosts
in the lifecycle. Plasmodium vivax, which infects humans, uses Anophele mosquitoes as a
vector. The parasite has a double-stranded DNA genome of around 27M nucleotides in
length spread over 14 chromosomes. This species is mainly found in Latin America and
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 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 gland ready to infect
another host when the mosquito feeds again.

After an incubation period varying between 7 and 30 days, infection with P. vivax 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. vivax
infections are most commonly treated with Primaquine which prevents the parasite being
released from the liver cells into the bloodstream of the host.

Introduction to Plasmodium vivax

2

MIN

MAX

The target sequence within the aspartic protease PM4 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 vivax (P.vivax) genomes is designed for the in
vitro quantification of P.vivax genomes. The kit is designed to have the broadest detection
profile possible whilst remaining specific to the P.vivax genome.

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

3

Kit Contents

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

FAM labelled

• P.vivax 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

4