PCRmax Leishmania major Instruction Manual

NADH dehydrogenase subunit 1
(ND1) kinetoplast mRNA

Leishmania major

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Leishmania major is a species of the Leishmania genus of parasitic organisms, which
cause zoonotic cutaneous leischmaniasis in humans. It is most prevalent in North Africa,
the Middle East, West Asia and Sudan. Its lifecycle encompasses two morphological
states; the promastigote is a motile flagellate that is approximately 10-20 µm in length
where as the amastigote is spheroid to ovoid in shape and are approximately 2.5- 5.0 µm
in length. It has a double stranded, linear DNA genome of approximately 32.8 Mb
comprising 36 chromosomes.
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L. major is an intracellular pathogen of the immune system targeting macrophages and
dendritic cells There are two hosts in the Leishmania’s protozoan life-cycle. The
intermediate host is the female sand-fly. In the mid-gut of the sand-fly, the parasite is a
promastigote that reproduces asexually and moves to the pharynx. From here it is expelled
into the skin of a vertebrate host when bitten. The parasite is initially phagocytosed before
spreading to blood and tissue cells of the host. Once within the host cell, the flagellum is
lost and the parasite, in its amastigote form, replicates by binary fission. The host cell
eventually bursts and the parasites are released to infect other cells throughout the body.

Cutaneous leishmaniasis can be identified by the large number of skin ulcers it produces
within few weeks of being bitten by female sand-flies. These ulcers are noticeably present
on the exposed part of the body such as the face, arms and legs. On healing, these ulcers
leave disabling scars. Patients suffering from this disease tend to develop secondary
infections that could potentially be fatal.

Introduction to Leishmania major

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MIN

MAX

Our kit for human Leishmania major has been designed for the specific and exclusive in
vitro quantification of this species and does not detect other species 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 Leishmania major (L.major) genomes is designed for the in
vitro quantification of L.major genomes. The kit is designed to have the broadest detection
profile possible whilst remaining specific to the L.major genome.

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

• L.major specific primer/probe mix (150 reactions BROWN)

FAM labelled

• L.major 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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