Swine flu is a viral infection caused by a zoonotic Influenza A virus strain similar to that found in pigs. In 2009 a strain of swine flu called H1N1 infected many people globally. The virus is contagious and can be spread between humans.
The A/H1N1pdm09 virus is now one of the seasonal flu viruses that circulate each winter. The symptoms are the same as other types of common flu. They’re usually mild and pass within 1 to 2 weeks. When people cough and sneeze, they spray tiny droplets of the virus into the air and if one comes into contact with these drops, they can catch the virus. People can spread it one day before they have any symptoms and as many as seven days after they are sick. The disease can also be transferred by eating bacon, ham or any other infected pork product. As with all types of flu, some people are at higher risk of serious illness, particularly those with underlying health problems.
The assay is an in vitro PCR reaction assay for the qualitative determination of 2009 H1N1 virus RNA in the human sample such nasopharyngeal swab and bronchoalveolar lavage (BAL) based on Taqman detection method for 2009 H1N1 virus with high sensitive one step RT-QPCR kit.
This kit detects the presence of H1pdm09 using one-step RT-qPCR (both reactions in the same tube) by first reverse transcribing the genomic RNA target to cDNA, followed by amplification of the assay target and detection by the hydrolysis probe method of qPCR. The assay consists of a forward primer, a reverse primer and a probe labelled with the 5’ FAM™ reporter dye and a 3’ quencher. As the new target cDNA strand is synthesised, the tightly bound probe is cleaved by the 5’ to 3’ exonuclease activity of Taq polymerase which releases the fluorescent reporter from the quencher and substantially increases the fluorescent signal. The point at which the fluorescence becomes detectable above the background, the quantification cycle (Cq), is proportional to the amount of target present in the sample. The lower the Cq, the greater the amount of target present. If, however Influenza A(H1N1) pdm09 is not present, a FAM signal will not be produced. An internal positive control assay is provided in order to assess the quality of the isolated RNA and the effect of any PCR inhibitors that may be present. This assay contains two primers and a HEX labelled probe, designed to a highly conserved region of human RNA and a positive signal indicates that the RNA quality in the sample is acceptable for diagnostic testing. These assays are both incorporated into a ready-to-use PCR master mix which utilises hot start technology, thus minimising non-specific reactions and ensuring maximum sensitivity.
The Zena Max H1N1 kit is sensitive down to a 55 copies per reaction under our validation methods and devices. The specificity of the AMD H1N1 assay is up to 100% for Influenza A Virus H1N1 of 2009 H1N1 Pandemic (Universal InfA, swine influenza A, swine H1 influenza and human RNase P gene as internal control) under our validation methods and devices.
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