PCRmax Haemophilus influenzae Instruction Manual
peptidoglycan-associated outer
membrane lipoprotein (OMP P6)
Haemophilus
influenzae
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
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Haemophilus influenzae is a Gram-negative bacterium of the Pasteurellaceae family. This
non-motile coccobacillus has a genome of around 1.8Mbp arranged in a single circular
conformation coding for 1740 genes and is around 1µm in length.
H. influenzae resides in the mucosal membrane of the nasopharynx and can be
transmitted via respiratory droplets. The bacteria colonise the nasopharynx via interactions
between outer membrane proteins and host mucin. Other bacterial proteins are then
expressed that impair ciliary activity, minimising the effect of the mucociliary escalator.
With bacteria within the mucus, a combination of pili and outer membrane proteins mediate
attachment to the host epithelial cells allowing colonisation. Once the bacteria have
colonised the nasopharynx, individuals invade the host cells and migrate between the
epithelial cells by disrupting tight junctions. The bacteria evade the host immune response
by expressing proteases that act on IgA present within the respiratory tract.
H. influenzae species can be classified into 2 types, encapsulated on nonencapsulated.
Nonencapsulated species often cause non-invasive infections. Of the encapsulated
strains, type b (Hib) is thought to be the most pathogenic although is only found in around
7% of the population whereas other strains can be found in around 75% of people.
Infection with Hib is most commonly seen in children and can cause bacteremia and acute
bacterial meningitis and less frequently epiglottitis, cellulitis and ear infections as well as
being involved in respiratory tract infections. Nonencapsulated strains are more frequently
seen in adults and can cause pneumonia. In 1988 a Hib vaccine was introduced which
has dramatically lowered the number of cases of Hib infections.
Introduction to Haemophilus influenzae
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MIN
MAX
The target sequence (outer membrane protein P6 gene) has previously been shown to be
a good genetic marker for H. influenza in other real time PCR based studies (Guma M.K
et.al 2009). 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 Haemophilus influenzae (H.influenzae ) genomes is designed
for the in vitro quantification of H.influenzae genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the H.influenzae genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
H.influenzae 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
• H.influenzae specific primer/probe mix (150 reactions BROWN)
FAM labelled
• H.influenzae 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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