PCRmax Brucella abortus Instruction Manual

outermembrane transporter
(PRK15319) gene

Brucella abortus

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Brucella abortus is an intracellular, blood-borne parasite. It is a Gram-negative
coccobacillus that causes an infectious and contagious disease called Brucellosis. The
disease primarily affects cattle but it can also be transmitted to humans from infected
animals and consuming their products. The disease can lead to great economic loss
especially in the dairy and agricultural industry. The Brucella abortus genome contains two
DNA chromosomes in a circular confirmation; the first chromosome is approximately 2.1
Mb and the second chromosome is approximately 1.2 Mb. Unusually it does not contain
any plasmids or genomic islands that relate to pathogenicity and lacks many other genes
that code for common virulence factors including capsules, fimbriae, exotoxins, cytolysins,
resistance forms, or antigenic variation.

The most common mode of transmission to humans is through the ingestion of
unpasteurized milk and cheese products as the bacteria are present in the milk glands of
infected female cows. In cattle transmission can also be through ingestion but in addition,
the bacteria can persist in the reproductive tracts of males, namely seminal vesicles,
ampullae, testicles, and epididymides, allowing sexual transmission. In humans the
bacteria enter macrophages by phagocytosis and then live in compartments of vacuolar
space along the endoplasmic reticulum. They persist by inhibiting host apoptosis and go
on to form chronic disease causing lesions in the liver, spleen, bone marrow and kidneys.
In cattle the bacteria additionally infect the trophoblast epithelial cells, which provide
nutrition to the embryo. The trophoblast cells eventually lyse, releasing further bacteria into
the blood stream of the embryo. The B. abortus cells in the blood stream go on to colonize
the placenta and fetus in pregnant female cows, resulting in abortion of the fetus. Abortion
can also result from insufficient anti-Brucella activity in the amniotic fluid.

In humans, the disease can be either acute or chronic and some of the symptoms include
fluctuating fever, chills, sweating, headache, muscle pain and weight loss. Once a person
becomes infected they are prescribed a combination of tetracycline and streptomycin for 3

-6 weeks. In cattle, additional symptoms include arthritic joints and retained after-birth.

Introduction to Brucella abortus

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MIN

MAX

The PCR Max qPCR Kit for Brucella abortus (B.abortus) genomes is designed for the in
vitro quantification of B.abortus genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the B.abortus genome.

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

• B.abortus specific primer/probe mix (150 reactions BROWN)

FAM labelled

• B.abortus 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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