IFNsig. type I Multiplex One Step RT-PCR Real Time
Quantitative analysis of Interferon Stimulated Genes ISG Cod. BM-023
Principle of the test : Quantitative analysis of Interferon Stimulated Genes ISG
Technology : Multiplex One Step RT- PCR Real Time
Gene Target : IFIT1 IFI44L IFI27 ISG15 RSAD2 SIGLEC
Internal Control: GAPDH RPS18
Specimen : RNA
Results : ΔΔCt method
Reporting Units : Arbitrary Units (AU)
Number of tests : 25 tests BM-023
Kit storage : -20°C
Necessary equipment : CFX96 Detection System, 7500 Real Time PCR System or other major Real-Time PCR platforms
Contacts:
cell: +39 329 0364389
e-Mail: info@biomole.it
biomole@ALL RIGHT RESERVED 2019
IFNsig. Type I One Step cod. BM-023 Multiplex One Step RT-PCR Real Time
Type I interferonopathies are characterized by an increase of circulating type I interferon (IFN) concentration. Type I interferonopathies refer to rare Mendelian genetic disorders such as Aicardi-Goutières Syndrome (AGS) as well as more frequent and polygenic auto-immune diseases like systemic lupus erythematosus (SLE). Moreover, in research laboratories this test is a valuable tool for predicting resistance to anti-cancer drugs, inflammatory diseases monitoring (such as inflammatory bowel diseases) and estimate the onset of autoimmune diseases in paediatric patients with thrombocytopenia.
Detection of type I IFN in these patients remains challenging as its amount is usually very low in patients' sera. Thus, the detection of interferon-stimulating genes has been proposed as an alternative for the detection of this cytokine but sensitivy, specificity and predictive values of the assay have not been reported so far.
The IFNsig.Type I Multiplex One Step kit is a research use only (RUO) real-time PCR assay able to reverse-transcribe and amplify RNA in a single step. Thanks to the multiplex version, the RNA is tested with two different mixes that will return the data of the six genes stimulated by Interferon α and the housekeeping gene. The relative quantification of the gene expression of these targets is performed with the ΔΔCt method and the fold change obtained is expressed in arbitrary units.
References:
1. Pescarmona R, Belot A, Villard M, Besson L, Lopez J, Mosnier I, Mathieu AL, Lombard C, Garnier L, Frachette C, Walzer T, Viel S. Comparison of RT-qPCR and Nanostring in the measurement of blood interferon response for the diagnosis of type I interferonopathies. Cytokine. 2018 Nov 6. pii: S1043-4666(18)30412-5. doi: 10.1016/j.CYto.2018.10.023
2. Tovo PA, Garazzino S, Daprà V, Pruccoli G, Calvi C, Mignone F, Alliaudi C, Denina M, Scolfaro C, Zoppo M, Licciardi F, Ramenghi U, Galliano I, Bergallo M. COVID-19 in Children: Expressions of Type I/II/III Interferons, TRIM28, SETDB1, and Endogenous Retroviruses in Mild and Severe Cases. Int J Mol Sci. 2021 Jul 13;22(14):7481. doi: 10.3390/ijms22147481.
3. Min Ae Lee-Kirsch. The Type I Interferonopathies. Annual Review of Medicine 2017;68:297-315
4. PIn A, Monasta L, Taddio A, Piscianz E, Tommasini A, Tesser A. An Easy and Reliable Strategy for Making Type I Interferon Signature Analysis Comparable among Research Centers. Diagnostics (Basel). 2019;9(3):113. doi: 10.3390/diagnostics9030113.
5. Kenneth J. Livak and Thomas D. Schmittgen. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2-[Delta][Delta]CT Method. Methods, 25(4):402 – 408, 2001
