Journal: Virology Journal

Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

doi: 10.1186/s12985-021-01559-3

Figure Lengend Snippet: CT value overview for 3600 clinical specimens tested with the RKI/ZBS1 SARS-CoV-2 protocol. CT values for the four PCR assays included in the RKI/ZBS1 SARS-CoV-2 protocol as determined for the first 3600 specimens. Red symbols show the SARS-CoV-2 assays (negatives not shown), blue squares the inhibition control KoMa and green circles the nucleic acid detection by c-myc amplification. The constant detection of the inhibition control KoMa can be seen, and the scattering of the nucleic acid content demonstrated by c-myc as well as the similar CT values for the E-Gene and the orf1ab assay

Article Snippet: The lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL) and the SOLIScript® 1-step CoV kit can amplify the KoMa control even in specimens highly positive for SARS-CoV-2, even with higher CT values than expected.

Techniques: Polymerase Chain Reaction, Inhibition, Amplification

Journal: Virology Journal

Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

doi: 10.1186/s12985-021-01559-3

Figure Lengend Snippet: RKI/ZBS1 SARS-CoV-2 protocol performance on different PCR cyclers. To show that the presented protocol can be run on several cyclers, we used 10 clinical samples of different RNA load, negative as well as positive controls and set up one master mix that was distributed to the different cyclers as shown above. Different colors represent 10 different samples used for comparison. Mean values of duplicates are shown, also for SARS-CoV-2-positive controls (crosses); negative controls are not shown

Article Snippet: The lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL) and the SOLIScript® 1-step CoV kit can amplify the KoMa control even in specimens highly positive for SARS-CoV-2, even with higher CT values than expected.

Techniques: Polymerase Chain Reaction

Journal: Virology Journal

Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

doi: 10.1186/s12985-021-01559-3

Figure Lengend Snippet: Correlation between the internal KoMa control and SARS-CoV-2 genome load in clinical specimens. For the 424 SARS-CoV-2-positive specimens the CT values were grouped as indicated and plotted against the CT value for the inhibition control KoMa. While in specimens that are highly positive for SARS-CoV-2 KoMa does not give a signal reliably, in specimens with genome loads close to the detection limit as well as in negative samples, KoMa is constantly amplified, showing the efficiency of the nucleic acid extraction and possible inhibitory effects of the sample matrix. Detectability increases significantly with E-Gene CT values higher than 25 (Mann Whitney p

Article Snippet: The lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL) and the SOLIScript® 1-step CoV kit can amplify the KoMa control even in specimens highly positive for SARS-CoV-2, even with higher CT values than expected.

Techniques: Inhibition, Amplification, MANN-WHITNEY

Journal: Virology Journal

Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

doi: 10.1186/s12985-021-01559-3

Figure Lengend Snippet: Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol

Article Snippet: The lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL) and the SOLIScript® 1-step CoV kit can amplify the KoMa control even in specimens highly positive for SARS-CoV-2, even with higher CT values than expected.

Techniques: Real-time Polymerase Chain Reaction

Journal: Virology Journal

Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

doi: 10.1186/s12985-021-01559-3

Figure Lengend Snippet: RKI/ZBS1 SARS-CoV-2 protocol performance with different PCR master mixes. In total 7 PCR mixes were compared with the RKI/ZBS1 SARS-CoV-2 protocol on a Bio-Rad CFX96 cycler. 10 clinical specimens of different RNA load and negative as well as positive controls were set up. Means of duplicate CT values are plotted against the respective master mix. Positive controls (crosses) are shown only for the SARS-CoV-2 assays E-Gene and orf1ab; negative controls are not shown

Article Snippet: The lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL) and the SOLIScript® 1-step CoV kit can amplify the KoMa control even in specimens highly positive for SARS-CoV-2, even with higher CT values than expected.

Techniques: Polymerase Chain Reaction

Journal: Virology Journal

Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

doi: 10.1186/s12985-021-01559-3

Figure Lengend Snippet: Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol

Article Snippet: To assess the performance of further commercially available one-step RT-PCR master mixes with the RKI/ZBS1 SARS-CoV-2 protocol, 10 clinical specimens with varying viral genome load were analyzed with the AgPath-ID master mix on the Bio-Rad CFX96 cycler and compared to the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase (Invitrogen, Carlsbad, CA, USA), the lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL, Berlin, Germany), the TaqPath™ 1-Step Multiplex Master Mix (No ROX) (Applied Biosystems), two kits from Solis BioDyne (SOLIScript 1-step Multiplex Probe Kit, SOLIScript® 1-step CoV Kit; Solis BioDyne, Tartu, Estonia) and the GoTaq® Probe 1-Step RT-qPCR System (Promega, Walldorf, Germany).

Techniques: Real-time Polymerase Chain Reaction