PCRmax Anaplasma phagocytophilum Instruction Manual
major surface protein 4 (msp4)
gene
Anaplasma
phagocytophilum
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
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Anaplasma phagocytophilum is a Gram-negative bacterium of the Anaplasma genus that is
responsible for causing human anaplasmosis, formerly known as human granulocytic
ehrlichiosis. It can also cause disease in dogs, ruminants, horses and sheep. It is an
obligate intracellular pathogen of 0.2-2 µm in diameter with a circular genome of
approximately 1.48 Mb. It is unable to synthesise peptidoglycan or lipopolysaccharide,
relying on membrane cholesterol to maintain physical integrity and leaving the organism
extremely vulnerable to stress.
A. phagocytophilum is usually transmitted through tick bites and the majority of cases
occur between April and September. The bacterium specifically attaches to neutrophils and
persist intracellularly within vacuoles. It produces a number of pathogenic factors that aid
virulence including specific adhesins for neutrophils and virulence factors that inhibit
phagosome-lysozome fusion and the production of reactive oxygen species that would
normally kill the bacterium. The bacterium also inhibits programmed cell death of the
neutrophil (apoptosis) and induces expression of interleukin-8, which causes neutrophil
chemotaxis, thereby increasing the spread of the bacterium throughout the host organism.
Symptoms include fever, shaking chills, myalgia, headache and nausea which typically
occur 7 to 10 days after tick bites.
Introduction to Anaplasma phagocytophilum
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MIN
MAX
The PCR Max qPCR Kit for Anaplasma phagocytophilum (A.phagocytophilum) genomes is
designed for the in vitro quantification of A.phagocytophilum genomes. The kit is designed
to have the broadest detection profile possible whilst remaining specific to the A.
phagocytophilum genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
A.phagocytophilum 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.phagocytophilum specific primer/probe mix (150 reactions BROWN)
FAM labelled
• A.phagocytophilum 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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