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201904151249411

IFNsig.type I-RT-PCR real time

Quantification assay of type I interferonopathies          cod. BM-005

 

Principle of the test: Quantitative analysis of ISGs mRNA expression

Technology: Relative Quantitative Real Time PCR

Target Genes: SIGLEC1, IFI27, IFI44L, ISG15, RSAD2, IFIT1.

Specimen: cDNA

Results: ΔΔCt method

Reporting Units: Arbitrary Units (AU)

Number of tests: 25 tests BM-005 

Kit storage: -20°C

Necessary equipment: Thermocycler, 7500 Real Time PCR System

Status: Ready to use

IFNsig.type I-RT-PCR real time cod. BM-005

 

Quantification assay of type I interferonopathies

 

·      IFNsig.type I-RT-PCR real time Quantification complete kit 25 tests BM-005

 

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). Yet, 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 (ISGs) has been proposed as an alternative for the detection of this cytokine but sensitivity, specificity and predictive values of the assay have not been reported so far9.

We developed a novel real-time qPCR assays able to amplified 6 ISGs that are among the highest ISGs expressed in AGS patients: SIGLEC1 (sialic acid binding Ig like lectin 1), IFI27 (interferon alpha inducible protein 27), IFI44L (Interferon induced protein 44 like), IFIT1 (interferon induced protein with tetratricopeptide repeats 1), ISG15 (interferon-stimulated gene 15) and RSAD2 (radical S-adenosyl methionine domain containing 2). Relative quantification of target gene expression in patients was performed with the ΔΔCt method and the relative ISGs fold changes were determined. Results are expressed in corresponding arbitrary units (AU) User friendly and complete, the IFNsig-RT-PCR real time Quantification kit is suitable for any laboratory.

 

Reference

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.