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agena bioscience ruo massarray sars cov 2 variant panel v1 agena bioscience

Workflow and molecular <t>principle</t> <t>of</t> <t>SARS-CoV-2</t> RNA detection in FFPE tissue by MALDI-TOF mass spectrometry. RNA extraction from FFPE placental and amniotic tissue was performed using the Maxwell RSC RNA FFPE Kit (Promega) according to manufacturer’s instructions. Extracted viral RNA is then reverse transcribed into cDNA and amplified by a one-step RT-PCR reaction. The figure schematically depicts specific amplification of the SARS-CoV-2 targets N1 and N2. The forward primers are indicated in green, the reverse primers in blue. Thereafter, PCR products are treated with the shrimp alkaline phosphatase (SAP) enzyme to remove unincorporated nucleotides (dNTPs). Next, a single nucleotide extension reaction is performed, in which target-specific extension primers are elongated by a single mass-modified terminator nucleotide (A, T, C or G) complementary to the cDNA template sequence. The extension products (EP) are then analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) using the <t>MassARRAY</t> system (Agena Bioscience) and mass spectra of the SARS-CoV-2-specific detected cDNA fragments are generated. In the absence of viral RNA, RT-PCR as well as single nucleotide extension reactions cannot be performed and consequently, no cDNA fragments are detected by MALDI-TOF analysis. EP: extension product; UEP: unextended extension primer.

Ruo Massarray Sars Cov 2 Variant Panel V1 Agena Bioscience, supplied by agena bioscience, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue"

Article Title: Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue

Journal: Viruses

doi: 10.3390/v14030604

Workflow and molecular principle of SARS-CoV-2 RNA detection in FFPE tissue by MALDI-TOF mass spectrometry. RNA extraction from FFPE placental and amniotic tissue was performed using the Maxwell RSC RNA FFPE Kit (Promega) according to manufacturer’s instructions. Extracted viral RNA is then reverse transcribed into cDNA and amplified by a one-step RT-PCR reaction. The figure schematically depicts specific amplification of the SARS-CoV-2 targets N1 and N2. The forward primers are indicated in green, the reverse primers in blue. Thereafter, PCR products are treated with the shrimp alkaline phosphatase (SAP) enzyme to remove unincorporated nucleotides (dNTPs). Next, a single nucleotide extension reaction is performed, in which target-specific extension primers are elongated by a single mass-modified terminator nucleotide (A, T, C or G) complementary to the cDNA template sequence. The extension products (EP) are then analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) using the MassARRAY system (Agena Bioscience) and mass spectra of the SARS-CoV-2-specific detected cDNA fragments are generated. In the absence of viral RNA, RT-PCR as well as single nucleotide extension reactions cannot be performed and consequently, no cDNA fragments are detected by MALDI-TOF analysis. EP: extension product; UEP: unextended extension primer.

Figure Legend Snippet: Workflow and molecular principle of SARS-CoV-2 RNA detection in FFPE tissue by MALDI-TOF mass spectrometry. RNA extraction from FFPE placental and amniotic tissue was performed using the Maxwell RSC RNA FFPE Kit (Promega) according to manufacturer’s instructions. Extracted viral RNA is then reverse transcribed into cDNA and amplified by a one-step RT-PCR reaction. The figure schematically depicts specific amplification of the SARS-CoV-2 targets N1 and N2. The forward primers are indicated in green, the reverse primers in blue. Thereafter, PCR products are treated with the shrimp alkaline phosphatase (SAP) enzyme to remove unincorporated nucleotides (dNTPs). Next, a single nucleotide extension reaction is performed, in which target-specific extension primers are elongated by a single mass-modified terminator nucleotide (A, T, C or G) complementary to the cDNA template sequence. The extension products (EP) are then analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) using the MassARRAY system (Agena Bioscience) and mass spectra of the SARS-CoV-2-specific detected cDNA fragments are generated. In the absence of viral RNA, RT-PCR as well as single nucleotide extension reactions cannot be performed and consequently, no cDNA fragments are detected by MALDI-TOF analysis. EP: extension product; UEP: unextended extension primer.

Techniques Used: RNA Detection, Mass Spectrometry, RNA Extraction, Reverse Transcription, Amplification, One Step RT-PCR, Modification, Sequencing, Generated, Reverse Transcription Polymerase Chain Reaction

Genomic targets of the MassARRAY SARS-CoV-2 Panels . ( A ): The CE-IVD-certified MassARRAY SARS-CoV-2 Panel targets 5 genomic regions within the SARS-CoV-2 genome: ORF1, ORF1ab, N1, N2 and N3 (indicated by red arrows). The detection of at least 2 targets is required for the result “SARS-CoV-2 positive”. ORF, open reading frame; S, spike; E, envelope; M, membrane; N, nucleocapsid. ( B ): The MassARRAY SARS-CoV-2 Variant Panel v1 targets 20 specific mutations (indicated by red arrows) within the spike gene allowing the identification of 5 different SARS-CoV-2 variants. For detection of the British (B.1.1.7) variant at least 6 out of the 8 mutations H69_70 del, Y144del, N501Y, A570D, P681H, T716I, S982A, D1118H are required while for detection of the South African (B.1.351) variant at least 6 out of the 8 mutations L18F, D80A, D215G, L242_L244del, K417N, E484K, N501Y and A701V are required. The 4 mutations L18F, K417T, E484K and N501Y identify the Brazilian (P.1) variant and the 4 mutations H69_70del, Y453F, N501T and I692V allow the identification of the Danish (Mink) variant (3 out of 4 mutations required). The mutation D614G evolved very early during the pandemic spread in 2020 and is present in all variants.

Figure Legend Snippet: Genomic targets of the MassARRAY SARS-CoV-2 Panels . ( A ): The CE-IVD-certified MassARRAY SARS-CoV-2 Panel targets 5 genomic regions within the SARS-CoV-2 genome: ORF1, ORF1ab, N1, N2 and N3 (indicated by red arrows). The detection of at least 2 targets is required for the result “SARS-CoV-2 positive”. ORF, open reading frame; S, spike; E, envelope; M, membrane; N, nucleocapsid. ( B ): The MassARRAY SARS-CoV-2 Variant Panel v1 targets 20 specific mutations (indicated by red arrows) within the spike gene allowing the identification of 5 different SARS-CoV-2 variants. For detection of the British (B.1.1.7) variant at least 6 out of the 8 mutations H69_70 del, Y144del, N501Y, A570D, P681H, T716I, S982A, D1118H are required while for detection of the South African (B.1.351) variant at least 6 out of the 8 mutations L18F, D80A, D215G, L242_L244del, K417N, E484K, N501Y and A701V are required. The 4 mutations L18F, K417T, E484K and N501Y identify the Brazilian (P.1) variant and the 4 mutations H69_70del, Y453F, N501T and I692V allow the identification of the Danish (Mink) variant (3 out of 4 mutations required). The mutation D614G evolved very early during the pandemic spread in 2020 and is present in all variants.

Techniques Used: Membrane, Variant Assay, Mutagenesis

Representative mass spectra of a positive sample analyzed by the CE-IVD-certified MassARRAY SARS-CoV-2 Panel. The different mass spectra display intensity and mass (in daltons) of the SARS-CoV-2 -specific cDNA fragments ORF1, ORF1ab, N1, N2 and N3. Additionally, the mass spectrum of the MS2 phage control is depicted. The red box highlights the unextended extension primer peak, whereas the blue box highlights the peak corresponding to the extension product. UEP: unextended extension primer; EP: extension product.

Figure Legend Snippet: Representative mass spectra of a positive sample analyzed by the CE-IVD-certified MassARRAY SARS-CoV-2 Panel. The different mass spectra display intensity and mass (in daltons) of the SARS-CoV-2 -specific cDNA fragments ORF1, ORF1ab, N1, N2 and N3. Additionally, the mass spectrum of the MS2 phage control is depicted. The red box highlights the unextended extension primer peak, whereas the blue box highlights the peak corresponding to the extension product. UEP: unextended extension primer; EP: extension product.

Techniques Used: Control

Detection of SARS-CoV-2 RNA in placental and amniotic tissue. Placental and amniotic tissue were analyzed by MALDI-TOF MS using the CE-IVD-certified MassARRAY SARS-CoV-2 Panel ( A ) and by RT-PCR using the EURORealTime SARS-CoV-2 Kit ( B ). ( A ): The plots display the intensity of the peaks corresponding to the 5 targets ORF1, ORF1ab, N1, N2 and N3 included in the CE-IVD-certified MassARRAY SARS-CoV-2 Panel. The intensity threshold considering a peak as detected is 5. All 5 SARS-CoV-2 targets were detected in both placental and amniotic tissue. ( B ). The graphs show the fluorescence intensity ( y -axis) for each cycle ( x -axis) of the real-time amplification of the SARS-CoV-2 -specific target regions (amplification curve). The amplification curve of the positive control (PC) is highlighted in green, the one of the negative control (NC) in red. The crossing point (Cp) value is indicated for both placental tissue (p.tissue) and amniotic tissue (a.tissue).

Figure Legend Snippet: Detection of SARS-CoV-2 RNA in placental and amniotic tissue. Placental and amniotic tissue were analyzed by MALDI-TOF MS using the CE-IVD-certified MassARRAY SARS-CoV-2 Panel ( A ) and by RT-PCR using the EURORealTime SARS-CoV-2 Kit ( B ). ( A ): The plots display the intensity of the peaks corresponding to the 5 targets ORF1, ORF1ab, N1, N2 and N3 included in the CE-IVD-certified MassARRAY SARS-CoV-2 Panel. The intensity threshold considering a peak as detected is 5. All 5 SARS-CoV-2 targets were detected in both placental and amniotic tissue. ( B ). The graphs show the fluorescence intensity ( y -axis) for each cycle ( x -axis) of the real-time amplification of the SARS-CoV-2 -specific target regions (amplification curve). The amplification curve of the positive control (PC) is highlighted in green, the one of the negative control (NC) in red. The crossing point (Cp) value is indicated for both placental tissue (p.tissue) and amniotic tissue (a.tissue).

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Fluorescence, Amplification, Positive Control, Negative Control

SARS-CoV-2 variant analysis of amniotic and placental tissue by MALDI-TOF MS. Placental and amniotic tissue were analyzed by MALDI-TOF MS using the MassARRAY SARS-CoV-2 Variant Panel v1. The figure shows the specific mutations within the SARS-CoV-2 spike gene that are assigned to the different variants B 1.1.7 ( A ), B 1.351 ( B ), P.1 ( C ) and Cluster 5/Mink ( D ). Detected mutations in placental tissue (p.tissue), amniotic tissue (a.tissue), positive controls (PC) B 1.1.7 and B 1.351 and negative control (NC) are illustrated by green-colored squares.

Figure Legend Snippet: SARS-CoV-2 variant analysis of amniotic and placental tissue by MALDI-TOF MS. Placental and amniotic tissue were analyzed by MALDI-TOF MS using the MassARRAY SARS-CoV-2 Variant Panel v1. The figure shows the specific mutations within the SARS-CoV-2 spike gene that are assigned to the different variants B 1.1.7 ( A ), B 1.351 ( B ), P.1 ( C ) and Cluster 5/Mink ( D ). Detected mutations in placental tissue (p.tissue), amniotic tissue (a.tissue), positive controls (PC) B 1.1.7 and B 1.351 and negative control (NC) are illustrated by green-colored squares.

Techniques Used: Variant Assay, Negative Control

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Journal: Viruses

Article Title: Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue

doi: 10.3390/v14030604

Figure Lengend Snippet: Workflow and molecular principle of SARS-CoV-2 RNA detection in FFPE tissue by MALDI-TOF mass spectrometry. RNA extraction from FFPE placental and amniotic tissue was performed using the Maxwell RSC RNA FFPE Kit (Promega) according to manufacturer’s instructions. Extracted viral RNA is then reverse transcribed into cDNA and amplified by a one-step RT-PCR reaction. The figure schematically depicts specific amplification of the SARS-CoV-2 targets N1 and N2. The forward primers are indicated in green, the reverse primers in blue. Thereafter, PCR products are treated with the shrimp alkaline phosphatase (SAP) enzyme to remove unincorporated nucleotides (dNTPs). Next, a single nucleotide extension reaction is performed, in which target-specific extension primers are elongated by a single mass-modified terminator nucleotide (A, T, C or G) complementary to the cDNA template sequence. The extension products (EP) are then analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) using the MassARRAY system (Agena Bioscience) and mass spectra of the SARS-CoV-2-specific detected cDNA fragments are generated. In the absence of viral RNA, RT-PCR as well as single nucleotide extension reactions cannot be performed and consequently, no cDNA fragments are detected by MALDI-TOF analysis. EP: extension product; UEP: unextended extension primer.

Article Snippet: For detection of the SARS-CoV-2 variants the Research Use Only (RUO) MassARRAY ® SARS-CoV-2 Variant Panel v1 (Agena Bioscience) was used.

Techniques: RNA Detection, Mass Spectrometry, RNA Extraction, Reverse Transcription, Amplification, One Step RT-PCR, Modification, Sequencing, Generated, Reverse Transcription Polymerase Chain Reaction

Journal: Viruses

Article Title: Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue

doi: 10.3390/v14030604

Figure Lengend Snippet: Genomic targets of the MassARRAY SARS-CoV-2 Panels . ( A ): The CE-IVD-certified MassARRAY SARS-CoV-2 Panel targets 5 genomic regions within the SARS-CoV-2 genome: ORF1, ORF1ab, N1, N2 and N3 (indicated by red arrows). The detection of at least 2 targets is required for the result “SARS-CoV-2 positive”. ORF, open reading frame; S, spike; E, envelope; M, membrane; N, nucleocapsid. ( B ): The MassARRAY SARS-CoV-2 Variant Panel v1 targets 20 specific mutations (indicated by red arrows) within the spike gene allowing the identification of 5 different SARS-CoV-2 variants. For detection of the British (B.1.1.7) variant at least 6 out of the 8 mutations H69_70 del, Y144del, N501Y, A570D, P681H, T716I, S982A, D1118H are required while for detection of the South African (B.1.351) variant at least 6 out of the 8 mutations L18F, D80A, D215G, L242_L244del, K417N, E484K, N501Y and A701V are required. The 4 mutations L18F, K417T, E484K and N501Y identify the Brazilian (P.1) variant and the 4 mutations H69_70del, Y453F, N501T and I692V allow the identification of the Danish (Mink) variant (3 out of 4 mutations required). The mutation D614G evolved very early during the pandemic spread in 2020 and is present in all variants.

Article Snippet: For detection of the SARS-CoV-2 variants the Research Use Only (RUO) MassARRAY ® SARS-CoV-2 Variant Panel v1 (Agena Bioscience) was used.

Techniques: Membrane, Variant Assay, Mutagenesis

Journal: Viruses

Article Title: Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue

doi: 10.3390/v14030604

Figure Lengend Snippet: Representative mass spectra of a positive sample analyzed by the CE-IVD-certified MassARRAY SARS-CoV-2 Panel. The different mass spectra display intensity and mass (in daltons) of the SARS-CoV-2 -specific cDNA fragments ORF1, ORF1ab, N1, N2 and N3. Additionally, the mass spectrum of the MS2 phage control is depicted. The red box highlights the unextended extension primer peak, whereas the blue box highlights the peak corresponding to the extension product. UEP: unextended extension primer; EP: extension product.

Article Snippet: For detection of the SARS-CoV-2 variants the Research Use Only (RUO) MassARRAY ® SARS-CoV-2 Variant Panel v1 (Agena Bioscience) was used.

Techniques: Control

Journal: Viruses

Article Title: Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue

doi: 10.3390/v14030604

Figure Lengend Snippet: Detection of SARS-CoV-2 RNA in placental and amniotic tissue. Placental and amniotic tissue were analyzed by MALDI-TOF MS using the CE-IVD-certified MassARRAY SARS-CoV-2 Panel ( A ) and by RT-PCR using the EURORealTime SARS-CoV-2 Kit ( B ). ( A ): The plots display the intensity of the peaks corresponding to the 5 targets ORF1, ORF1ab, N1, N2 and N3 included in the CE-IVD-certified MassARRAY SARS-CoV-2 Panel. The intensity threshold considering a peak as detected is 5. All 5 SARS-CoV-2 targets were detected in both placental and amniotic tissue. ( B ). The graphs show the fluorescence intensity ( y -axis) for each cycle ( x -axis) of the real-time amplification of the SARS-CoV-2 -specific target regions (amplification curve). The amplification curve of the positive control (PC) is highlighted in green, the one of the negative control (NC) in red. The crossing point (Cp) value is indicated for both placental tissue (p.tissue) and amniotic tissue (a.tissue).

Article Snippet: For detection of the SARS-CoV-2 variants the Research Use Only (RUO) MassARRAY ® SARS-CoV-2 Variant Panel v1 (Agena Bioscience) was used.

Techniques: Reverse Transcription Polymerase Chain Reaction, Fluorescence, Amplification, Positive Control, Negative Control

Journal: Viruses

Article Title: Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) including Variant Analysis by Mass Spectrometry in Placental Tissue

doi: 10.3390/v14030604

Figure Lengend Snippet: SARS-CoV-2 variant analysis of amniotic and placental tissue by MALDI-TOF MS. Placental and amniotic tissue were analyzed by MALDI-TOF MS using the MassARRAY SARS-CoV-2 Variant Panel v1. The figure shows the specific mutations within the SARS-CoV-2 spike gene that are assigned to the different variants B 1.1.7 ( A ), B 1.351 ( B ), P.1 ( C ) and Cluster 5/Mink ( D ). Detected mutations in placental tissue (p.tissue), amniotic tissue (a.tissue), positive controls (PC) B 1.1.7 and B 1.351 and negative control (NC) are illustrated by green-colored squares.

Article Snippet: For detection of the SARS-CoV-2 variants the Research Use Only (RUO) MassARRAY ® SARS-CoV-2 Variant Panel v1 (Agena Bioscience) was used.

Techniques: Variant Assay, Negative Control

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