PCRmax Leishmania infantum and L. donovani Instruction Manual

DNA polymerase I protein B
(POLIB) gene

Leishmania infantum &
Leishmania donovani

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Leishmania infantum (L.infantum) is an intracellular protozoan parasite that is part of the
genus Leishmania and part of the L.donovani complex. It is responsible for causing
visceral leishmaniasis. The genome consists of 36 chromosomes with a total genome size
of just over 32Mb. Like all members of the Kinetoplastids class, aside from the DNA carried
on the chromosomes, they also carry kinetoplast circle DNA within the mitchondria.
Depending on the host, the morphological features of the organism vary. Within
mammalian host, the protozoan is in the form of an amistigote, a round non-motile form
that is only 3-7 micrometres in diameter. Within the sandfly vector, the amastigote
transforms into a promastigote, a spindle shaped cell that is 3 times larger and has a single
flagellum which allows for motility.

Leishmania donovani (L.donovani) is also responsible for causing visceral leishmaniasis
and is closely related to L.infantum to the point where some authors believe that these two
species are subspecies of each other.

The pathogen is spread by spread through an insect vector, the sandflies of the
Phlebotomus and the Lutzomyia genus. Once inside the mammalian host, promastigotes
invade macrophages. Once inside, they transform back into the smaller amastigote form
and reproduce within the cell, eventually causing it to lyse. The released amistigotes then
infect other macrophage cells and in time, becomes a systemic infection targeting the
spleen and liver.

Typical symptoms are fever and the enlargement of the spleen, or splenomegaly, with
enlargement of the liver, hepatomegaly and is fatal if it is left untreated. Post-kala-azar
dermal leishmaniasis (PKDL) is characterized by a macular, maculo-papular or nodular
rash and is a complication of VL that is frequently observed after treatment

Introduction to Leishmania infantum & Leishmania
donovani

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MIN

MAX

The PCR Max qPCR Kit for Leishmania infantum & Leishmania donovani (L.infantum/L.
donovani) genomes is designed for the in vitro quantification of L.infantum/L.donovani
genomes. The kit is designed to have the broadest detection profile possible whilst
remaining specific to the L.infantum/L.donovani genome.

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

FAM labelled

• L.infantum/L.donovani 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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