PCRmax H1N1 Influenza Instruction Manual
M1 - global Influenza A
&
N1- specific for Swine H1N1
Influenza – Human Pandemic
Strain
Swine H1N1 Influenza
Human Pandemic
Strain
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
Quantification of Swine H1N1 Influenza Human Pandemic Strain genomes
Advanced kit handbook HB10.05.09
Published Date: 26/04/2016
1
Influenza, commonly known as the flu, is an infectious disease of birds and mammals
caused by an RNA virus of the family Orthomyxoviridae (the influenza viruses). In people,
common symptoms of influenza are fever, sore throat, muscle pains, severe headache,
coughing, and weakness and fatigue. In more serious cases, influenza causes
pneumonia, which can be fatal, particularly in young children and the elderly. Although the
common cold is sometimes confused with influenza, it is a much less severe disease and
caused by a different virus. Similarly, gastroenteritis is sometimes called "stomach flu" or
"24-hour flu", but is unrelated to influenza.
Typically, influenza is transmitted from infected mammals through the air by coughs or
sneezes creating aerosols containing the virus, and from infected birds through their
droppings. Influenza can also be transmitted by saliva, nasal secretions, feces and blood.
Infections either occur through direct contact with these bodily fluids, or by contact with
contaminated surfaces. Flu viruses can remain infectious for over 30 days at 0°C (32°F)
and about one week at human body temperature, although they are rapidly inactivated by
disinfectants and detergents.
Flu spreads around the world in seasonal epidemics, killing millions of people in pandemic
years and hundreds of thousands in non-pandemic years. Three influenza pandemics
occurred in the 20th century – each following a major genetic change in the virus – and
killed tens of millions of people. Often, these pandemics result from the spread of a flu
virus between animal species. Since it first killed humans in Asia in the 1990s a deadly
avian strain of H5N1 has posed the greatest influenza pandemic threat. However, this virus
has not yet mutated to spread easily between people.
Vaccinations against influenza are most common in high-risk humans in industrialised
countries and farmed poultry. The most common human vaccine is the trivalent flu vaccine
that contains purified and inactivated material from three viral strains. Typically this vaccine
includes material from two influenza A virus subtypes and one influenza B virus strain. A
vaccine formulated for one year may be ineffective in the following year, since the
Influenza A Virus changes every year and different strains become dominant. Antiviral
drugs can be used to treat influenza, with neuraminidase inhibitors being particularly
effective.
Introduction to Swine H1N1 Influenza Human
Pandemic Strain
Quantification of Swine H1N1 Influenza Human Pandemic Strain genomes
Advanced kit handbook HB10.05.09
Published Date: 26/04/2016
2
Specificity
The kit has 2 primer and probe sets. The M1 primers and probe detect ALL influenza A
isolates including those originally isolated from both human and swine. The primers have
100% homology to all pandemic H1N1 sequences.
The N1 primer and probe set detects ONLY the pandemic swine flu H1N1. The primers
and probe have 100% homology to all pandemic strains. The primers will not detect N1
from any seasonal endemic human or swine N1 strains.
The PCRmax qPCR Kit for Swine H1N1 Influenza Human Pandemic Strain (H1N1-swine)
genomes is designed for the in vitro quantification of H1N1-swine genomes. The kit is
designed to have the broadest detection profile possible whilst remaining specific to the
H1N1-swine genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
H1N1-swine 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.
Quantification of Swine H1N1 Influenza Human Pandemic Strain genomes
Advanced kit handbook HB10.05.09
Published Date: 26/04/2016
3
Kit Contents
• M1 global influenza A primer/probe mix (150 reactions BROWN)
FAM labelled
• N1 swine H1N1 influenza specific primer/probe mix (150 reactions BROWN)
FAM labelled
• M1 global influenza A positive control template (for Standard curve RED)
• N1 swine H1N1 influenza specific positive control template (for Standard curve
RED)
• Internal extraction control primer/probe mix (150 reactions BROWN)
VIC labelled as standard
• Internal extraction control RNA (150 reactions BLUE)
• Endogenous control primer/probe mix (150 reactions BROWN)
FAM labelled
• M1 and N1/Internal extraction control/endogenous control RT primer mix (150
reactions GREEN)
Required for two step protocol only
• RNAse/DNAse free water (WHITE)
for resuspension of primer/probe mixes and internal extraction control RNA
• Template preparation buffer (YELLOW)
for resuspension of positive control templates and standard curve preparation
Reagents and equipment to be supplied by the user
Real-Time PCR Instrument
RNA extraction kit
This kit is designed to work well with all processes that yield high quality RNA with
minimal PCR inhibitors.
Lyophlised 2x RT-qPCR MasterMix
This kit is designed to be compatible with all commercially available OneStep
Mastermixes that run with standard cycling conditions.
Pipettors and Tips
Vortex and centrifuge
Thin walled 1.5 ml PCR reaction tubes
Quantification of Swine H1N1 Influenza Human Pandemic Strain genomes
Advanced kit handbook HB10.05.09
Published Date: 26/04/2016
4
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