Alpha-1-AT PCR Assay
The Viennalab AAT mpx RealFast Assay detects the two deficiency variants of the SERPINA1 gene in one multiplex real-time PCR run: PI*S (g.14768A>T, rs17580) and PI*Z (g.17083G>A, rs28929474). SERPINA1 encodes alpha-1-antitrypsin, the protease inhibitor that protects lung tissue against neutrophil elastase. Carriers of the Z allele, in particular the ZZ genotype, have strongly reduced plasma levels and are at risk of emphysema and of liver disease. The S allele gives a milder reduction and matters mainly in combination with Z.
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This kit in detail
- Simultaneous detection of the PI*S and PI*Z variants of SERPINA1
- Multiplex real-time PCR, amplicons of 145 bp and 153 bp
- Four allele-specific probes: FAM and HEX for S, ROX and Cy5 for Z
- WT-Control and MUT-Control supplied
- Whole peripheral blood, dried blood spots, buccal swabs or saliva
- 32 or 100 reactions per kit, REF 7-268 and REF 7-265
The Viennalab AAT mpx RealFast Assay determines the genotype of the two protease inhibitor variants PI*S and PI*Z in the SERPINA1 gene, which together account for most cases of alpha-1-antitrypsin deficiency.
Clinical background
Alpha-1-antitrypsin inhibits neutrophil elastase and so protects the alveolar wall against proteolytic damage. The Z variant causes the protein to polymerise in the hepatocyte, which both lowers the plasma level and loads the liver. Homozygous ZZ carriers develop early emphysema, in smokers earlier still, and can develop liver disease in childhood or later in life. The S variant lowers the level less, and is relevant mainly as the SZ compound heterozygote. The genotype supports the interpretation of a low alpha-1-antitrypsin concentration and the counselling of family members.
Method
The assay amplifies fragments of 145 bp and 153 bp of the SERPINA1 gene in one reaction and reads four allele-specific hydrolysis probes: FAM for the S allele and HEX for the corresponding M allele, ROX for the Z allele and Cy5 for the corresponding M allele. The kit supplies the RealFast 2x mpx Probe Mix, the AAT mpx Assay Mix, a WT-Control and a MUT-Control. The recommended DNA input is 10 to 100 ng per reaction, at a concentration of 2 to 20 ng/µl. DNA isolated from whole peripheral blood, dried blood spots, buccal swabs or saliva can be used, and DNA extraction reagents are not supplied with the kit.
Instrument compatibility
Because the assay uses the ROX channel for one of its probes, it cannot be run on instruments that need ROX for normalisation: the Applied Biosystems StepOne, 7300 and 7900/7900HT are not suitable. The IFU lists the AB 7500 Fast, Bio-Rad CFX96, Roche LightCycler 480, bms MIC qPCR Cycler and Qiagen Rotor-Gene 6000 as suitable.
- RealFast™ 2x mpx Probe Mix
- AAT mpx Assay Mix
- AAT mpx WT-Control
- AAT mpx MUT-Control
Datasheets and regulatory documentation
Everything your quality team needs for evaluation and procurement.
Instructions for Use
Frequently paired with this kit
Other assays and instruments lab teams add alongside the Alpha-1-AT PCR Assay.
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