PCRmax Trypanosoma cruzi Instruction Manual

18S ribosomal RNA (18S) gene

Trypanosoma cruzi

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Trypanosoma cruzi is a parasitic protozoan which is the causative agents of American
Trypanosomiasis or Chagas disease mainly found in Central and Southern America. The
T.cruzi genome is 67Mb in length spread over 41 chromosome pairs ranging in size from
78Kb to 2.4Mb. this species has a single nucleus and also a kinetoplast. Genome analysis
has shown there to be two distinct lineages of this species with one infecting marsupials
and the other infecting placental mammals.

T.cruzi has a two stage life cycle utilising hematophagous triatomine insects such as
Triatoma infestans as a vector to infect vertebrate hosts. In the vector the epimastigote
stage of the parasite reproduce through binary fission and moves into the rectal cell wall of
the insect. While feeding on the host the triatomine insect passes the infectious stage of
the parasite (metacyclic trypomatsigotes) through faeces which then enters the host via
open wounds or mucosal membranes. The trypomastigotes transforms into amastigotes
inside the host tissue and back into trypomastigotes bursting out of the cell to enter the
blood stream.

The most recognized symptom of Trypanosomiasis is swelling of one eye but other
symptoms include tiredness, fever, swelling of the lymph glands, enlarged liver or spleen.
Some patients develop irreversible damage to the heart and the intestinal tract appearing
many years after initial infection. The chronic form of the illness is associated with the
upregulation of phosphorylated extra cellular –signal-regulated kinase and the activation of
Cyclin D1, a mitotic regulator for G1 progression.

Introduction to Trypanosoma cruzi

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MIN

MAX

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

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

• T.cruzi specific primer/probe mix (150 reactions BROWN)

FAM labelled

• T.cruzi 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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