oasig lyophilised OneStep qRT-PCR MasterMix

RNAemia levels were measured by a One-Step qRT-PCR detection kit (Oasig, Primerdesign Ltd., UK), and using a DENV RT primer/probe Mix kit (Genesig, Primerdesign Ltd., UK) according to the manufacturer’s protocol (catalog no. oasig-onestep).

Pérez-Guzmán, E.X., Pantoja, P., Serrano-Collazo, C. et al. Time elapsed between Zika and dengue virus infections affects antibody and T cell responses. Nat Commun 10, 4316 (2019).

https://doi.org/10.1038/s41467-019-12295-2

Time elapsed between Zika and dengue virus infections affects antibody and T cell responses | Nature Communications

Measured the genomic copy number by quantitative RT-PCR, using the Advanced kit for Tick-borne encephalitis (Genesig) and Lyophilised OneStep qRT-PCR (Oasig) following manufactures manufacturer’s instructions.

Svoboda P, Haviernik J, Bednar P, Matkovic M, Cervantes Rincón T, Keeffe J, Palus M, Salat J, Agudelo M, Nussenzweig MC, Cavalli A, Robbiani DF, Ruzek D. A combination of two resistance mechanisms is critical for tick-borne encephalitis virus escape from a broadly neutralizing human antibody. Cell Rep. 2023 Sep 26;42(9):113149. doi: 10.1016/j.celrep.2023.113149. Epub 2023 Sep 19. PMID: 37715951; PMCID: PMC10591882.

A combination of two resistance mechanisms is critical for tick-borne encephalitis virus escape from a broadly neutralizing human antibody - PMC (nih.gov)

The genesig real-time PCR detection kit from Primerdesign™ Ltd. supplied by oasig lyophilized One-Step RT-qPCR MasterMix was used for HEV detection (based on the detection of the ORF2 capsid protein-encoding gene).

Bazir H, Hassou N, El Mellouli F, Zekhnini H, Najib S, Ennaji MM. Hepatitis A and E Viruses in Mussels from Cherrat Estuary in Morocco: Detection by Real-Time Reverse Transcription PCR Analysis. Adv Virol. 2022 Nov 28;2022:8066356. doi: 10.1155/2022/8066356. PMID: 36479562; PMCID: PMC9722280.

Hepatitis A and E Viruses in Mussels from Cherrat Estuary in Morocco: Detection by Real-Time Reverse Transcription PCR Analysis - PMC (nih.gov)

Real-time PCR was carried out by using a Bio-Rad CFX96 Touch System™. The amplification was performed by using commercial kits: for Herpes simplex virus (HSV 1 & 2), the herpes simplex virus 1 and 2 DNA polymerase (UL30) gene - genesig standard kit; for Cytomegalovirus (HCMV), the human betaherpes virus 5 - cytomegalovirus, glycoprotein B (gB) gene - genesig standard kit, and for Epstein Barr virus (EBV), the Epstein Barr virus (human herpes virus 4) nonglycosylated membrane protein (BNRF1) - gene genesig advanced kit, which included a positive control template. We used Oasig lyophilised 2X qPCR MasterMixTM.

Sánchez-Villamil JP, Pino-Vélez C, Trejos-Suárez J, Cardona N, España AL, Alfonso PA. Salivary markers of oxidative stress and periodontal pathogens in patients with periodontitis from Santander, Colombia. Biomedica. 2020 May 1;40(Supl. 1):113-124. doi: 10.7705/biomedica.5149. PMID: 32463613; PMCID: PMC7449106.

Salivary markers of oxidative stress and periodontal pathogens in patients with periodontitis from Santander, Colombia - PMC (nih.gov)

EBOV was quantified in PBMC RNA by using the Ebola 2014 outbreak genesig qRT-PCR kit (which detects the EBOV Makona strain nucleoprotein gene) and oasig™ one-step qRT-PCR master mix (Primerdesign) according to the manufacturer’s instructions.

Eisfeld AJ, Halfmann PJ, Wendler JP, Kyle JE, Burnum-Johnson KE, Peralta Z, Maemura T, Walters KB, Watanabe T, Fukuyama S, Yamashita M, Jacobs JM, Kim YM, Casey CP, Stratton KG, Webb-Robertson BM, Gritsenko MA, Monroe ME, Weitz KK, Shukla AK, Tian M, Neumann G, Reed JL, van Bakel H, Metz TO, Smith RD, Waters KM, N'jai A, Sahr F, Kawaoka Y. Multi-platform 'Omics Analysis of Human Ebola Virus Disease Pathogenesis. Cell Host Microbe. 2017 Dec 13;22(6):817-829.e8. doi: 10.1016/j.chom.2017.10.011. Epub 2017 Nov 16. PMID: 29154144; PMCID: PMC5730472.

Multi-Platform ‘Omics Analysis of Human Ebola Virus Disease Pathogenesis - PMC (nih.gov)

Viral copy number in approximately 100 ng nucleic acid extract was determined on a QuantStudio 5 real-time PCR System (Applied Biosystems, Thermo Fisher Scientific) using the GeneSig Standard CHIKV qRT-PCR assay with oasig One Step qRT-PCR master mix (PrimerDesign) according to the manufacturer’s instructions.

Rayner JO, Kim JH, Roberts RW, Wood RR, Fouty B, Solodushko V. Evaluation of DNA-Launched Virus-Like Particle Vaccines in an Immune Competent Mouse Model of Chikungunya Virus Infection. Vaccines (Basel). 2021 Apr 2;9(4):345. doi: 10.3390/vaccines9040345. PMID: 33918409; PMCID: PMC8067036.

Evaluation of DNA-Launched Virus-Like Particle Vaccines in an Immune Competent Mouse Model of Chikungunya Virus Infection - PMC (nih.gov)

In parallel, to confirm the PCR result, real-time RT-PCR to detect the RNA of avian orthoreovirus Nonstructural Protein σ was performed using an Avian Orthoreovirus Advanced kit (Genesig, Primerdesign, Chandler’s Ford, UK) and oasig lyophilized OneStep RT-qPCR Master Mix (Genesig, Primerdesign, Chandler’s Ford, UK) according to the manufacturer’s instructions, and Applied Biosystems 7500 Fast Real-Time PCR System.

Nowak T, Kwiecinski A, Kwiecinski P, Tomczyk G, Wodz K. Detection and Identification of Avian Reovirus in Young Geese (Anser anser domestica) in Poland. Animals (Basel). 2022 Nov 29;12(23):3346. doi: 10.3390/ani12233346. PMID: 36496863; PMCID: PMC9736766.

Detection and Identification of Avian Reovirus in Young Geese (Anser anser domestica) in Poland - PMC (nih.gov)

Each reaction mixture (20 µl) contained 10 µl oasig™ lyophilised OneStep qRT-PCR MasterMix (PrimerDesign, Ltd.), 1 µl primer/probe mix, 1 µl internal extraction control primer/probe mix, 3 µl RNse-free water, and 5 µl of RNA sample.

Stobnicka A, Gołofit-Szymczak M, Wójcik-Fatla A, Zając V, Korczyńska-Smolec J, Górny RL. Prevalence of Human Parainfluenza Viruses and Noroviruses Genomes on Office Fomites. Food Environ Virol. 2018 Jun;10(2):133-140. doi: 10.1007/s12560-017-9327-z. Epub 2017 Dec 1. PMID: 29196954; PMCID: PMC5951877.

Prevalence of Human Parainfluenza Viruses and Noroviruses Genomes on Office Fomites - PMC (nih.gov)

RV2 or RSVA2 intracellular RNA was reverse transcribed using RV or RSV-specific primers and cDNA amplified using RV or RSV TaqMan® primer-probe mix and oasig™ onestep qRT-PCR mastermix (Primerdesign).

Baturcam E, Vollmer S, Schlüter H, Maciewicz RA, Kurian N, Vaarala O, Ludwig S, Cunoosamy DM. MEK inhibition drives anti-viral defence in RV but not RSV challenged human airway epithelial cells through AKT/p70S6K/4E-BP1 signalling. Cell Commun Signal. 2019 Jul 18;17(1):78. doi: 10.1186/s12964-019-0378-7. PMID: 31319869; PMCID: PMC6639958.

MEK inhibition drives anti-viral defence in RV but not RSV challenged human airway epithelial cells through AKT/p70S6K/4E-BP1 signalling - PMC (nih.gov)