PCRmax Anaplasma marginale Instruction Manual
major surface protein 4 (msp 4)
gene
Anaplasma marginale
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
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Anaplasma marginale is a Gram-negative bacterium of the Rickettsiales order lacking a
traditional cell wall. The double-stranded, circular genome of this species is around 1.2Mb
in length coding for over 900 proteins. Infection with this species is a major threat to livestock and causes anaplasmosis.
A. Marginale use ticks such as Boophilus spp. and Dermacentor spp as a vector and
multiply within them before being passed to cattle when the ticks feed. The bacteria infect
the erythrocytes of the cattle by production of endotoxins and multiply within the cells
mainly in the spleen and liver. The host immune system destroys infected cells leading to
anaemia.
Cattle infection presents with fever, loss of appetite, jaundice and respiratory problems.
The infected animal may recover from the clinical symptoms of the infection but will remain
a carrier of the bacteria, passing it back to ticks during feeding and also be immune to
future infections. Immunity can be passed to calves via milk feeding and offers protection
to infants. In severe case, the infection may result in animal fatality.
Blood-based live vaccines are often used in tropical countries to protect animals but due to
the risks associated with transmission of other diseases, these vaccines are not available
in many countries.
Introduction to Anaplasma marginale
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MIN
MAX
The PCR Max qPCR Kit for Anaplasma marginale (A.marginale) genomes is designed for
the in vitro quantification of A.marginale genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the A.marginale genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
A.marginale 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 help@pcrmax.com and our bioinformatics team will
answer your question.
Specificity
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Kit Contents
• A.marginale specific primer/probe mix (150 reactions BROWN)
FAM labelled
• A.marginale 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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