PCRmax Aspergillus fumigatus Instruction Manual

hypothetical protein
(AFUA_3G08890)

Aspergillus fumigatus

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Aspergillus fumigatus is a pathogenic fungus of the genus Aspergillus. It is a saprotroph
that is found widely in nature, typically in soil and decaying organic matter, such as
compost heaps, and is the causative agent for the disease aspergillosis. A. fumigatus has
a haploid 29.4 Mb genome that forms eight chromosomes. It forms colonies that produce
hyphae known as conidiophores that produce thousands of minute grey-green conidia (2–3
μm) that readily become airborne. Inhaling the spores can result in allergic
bronchopulmonary aspergillosis, pulmonary aspergilloma or invasive aspergillosis.

Infection occurs after inhalation and it is estimated that everybody inhales several hundred
spores each day; typically these are quickly eliminated by the immune system in healthy
individuals. The fungus is more likely to become pathogenic in immunocompromised
individuals. Due to the small size of conidia, many can become deposited in alveoli where
they interact with innate effecter cells, usually leading to their destruction. If they evade the
immune system they emerge from dormancy and make a morphological switch to hyphae
after germinating in the pulmonary alveoli. Germination occurs either extracellularly or in
type II pneumocyte endosomes. Following germination, filamentous hyphal growth results
in epithelial penetration and subsequent penetration of the vascular endothelium. This
causes cellular damage and induces repair mechanisms, including a proinflammatory
response, which results in intravascular thrombosis and localized tissue necrosis.
Dissemination through the blood stream only occurs in severely immunocomprimised
individuals and can affect major organs including the brain, heart and kidneys.

Symptoms vary depending on the severity of the infection. A fungus ball in the lung may be
asymptomatic, may cause repeated coughing up of blood or occasionally severe, even
fatal, bleeding. Invasive aspergillosis of the lungs often causes coughing, fever, chest pain
and difficulty breathing. When deeper tissues are affected symptoms include fever, chills,
shock, delirium, and blood clots. The person may develop kidney failure, liver failure and
breathing difficulties. Death can occur quickly.

Introduction to Aspergillus fumigatus

2

MIN

MAX

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

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

3

Kit Contents

• A.fumigatus specific primer/probe mix (150 reactions BROWN)

FAM labelled

• A.fumigatus 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

4