Liaison mdx instrument

Manufactured by DiaSorin

The LIAISON® MDX instrument is a fully automated, random-access analyzer designed for clinical laboratory testing. It is capable of performing a wide range of immunoassay tests, including those for infectious diseases, endocrinology, and specialty analytes. The LIAISON® MDX instrument utilizes chemiluminescent technology to provide accurate and reliable results.

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Lab products found in correlation

Simplexa covid 19 direct kit, by DiaSorin (1 mentions) Simplexa covid 19 direct assay, by DiaSorin (1 mentions)

6 protocols using liaison mdx instrument

SARS-CoV-2 RNA Detection Protocol

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All 118 NPS were routinely tested for the presence of SARS-CoV-2 RNA using the Simplexa^TM SARS-CoV-2 Direct assay.
The run was performed by using a Direct Amplification Disk, consisting of eight simultaneous sample reactions. For the amplification, we used 50 μl of the mix per reaction and 50 μl for each sample, loaded onto the LIAISON® MDX instrument (DiaSorin S.p.A., Gerenzano, Italy).
Samples with cycle threshold (Ct) values <40 were considered positive; for statistical calculations, an arbitrary value of 45 Ct was assigned to negative samples.

Sberna G., Fabeni L., Berno G., Carletti F., Specchiarello E., Colavita F., Meschi S., Matusali G., Garbuglia A.R., Bordi L, & Lalle E. (2022). Rapid and qualitative identification of SARS-CoV-2 mutations associated with variants of concern using a multiplex RT-PCR assay coupled with melting analysis. International Journal of Infectious Diseases, 122, 401-404.

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SARS-CoV-2 Genome Detection Assay

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This assay is designed to detect the ORF1ab and S regions of the SARS-CoV-2 genome. Briefly, 50 μL of sample was transferred into the direct amplification disk (DAD) in addition to 50 μL of reaction mix. The sealed DAD was loaded into the LIAISON® MDX instrument (DiaSorin Molecular LLC) and run using the COVID-19 protocol. This assay requires detection of only one of the gene targets to be considered positive.

Kanwar N., Banerjee D., Sasidharan A., Abdulhamid A., Larson M., Lee B., Selvarangan R, & Liesman R.M. (2021). Comparison of diagnostic performance of five molecular assays for detection of SARS-CoV-2. Diagnostic Microbiology and Infectious Disease, 101(4), 115518.

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COVID-19 RT-PCR Detection Protocol

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Samples were processed and analyzed according to manufacturer’s instructions. Prior to batch analysis, NPS specimens were inactivated at 56 °C for 30 min and stored at −80 °C for less than one week. RT-PCR was performed on 50 µL of preprocessed VTM using the Simplexa™ COVID-19 Direct RT-PCR assay on the LIAISON^® MDX instrument (DiaSorin). This RT-PCR assay targets both spike (S) and ORF1ab viral genes. The latter, ORF1ab gene target, was used as reference because of a better sensitivity. However, the crossing threshold (Ct) values were recorded for both targets (S and ORF1ab). Higher Ct values represent a low viral RNA load.

Petonnet D., Marot S., Leroy I., Cohier J., Ramahefasolo C., Mansaly S., Calvez V., Marcelin A.G, & Burrel S. (2022). Comparison of Rapid and Automated Antigen Detection Tests for the Diagnosis of SARS-CoV-2 Infection. Diagnostics, 12(1), 104.

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Real-time PCR Assay for P. jirovecii

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The DiaSorin P. jirovecii real-time PCR assay (DiaSorin Molecular LLC, Cypress, USA) was setup according to manufacturer’s instructions. Briefly, 0.2 μL DiaSorin P. jirovecii primer pair was mixed with 4 μL temperature activated (TA) master mix, 0.2 μL Simplexa extraction and amplification control DNA, 0.2 μL Simplexa extraction and amplification control primer pair (DiaSorin), and 3.4 μL nuclease-free water. Subsequently, 8 μL of this reaction mixture was added to each well of a 96-well Simplexa Universal disk (DiaSorin), and 2 μL of unprocessed, neat respiratory specimen was directly added to each well inside a biosafety cabinet. PCRs were performed on the Liaison MDX instrument (DiaSorin) using the manufacturer recommended thermocycling program and the liaison MDX Studio Software for data analysis.

Belanger C.R., Locher K., Velapatino B., Dufresne P.J., Eckbo E, & Charles M. (2023). Quick versus Quantitative: Evaluation of Two Commercial Real-Time PCR Assays for the Detection of Pneumocystis jirovecii from Bronchoalveolar Lavage Fluids. Microbiology Spectrum, 11(4), e01021-23.

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Quantitative SARS-CoV-2 Detection in Nasal Wash

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Quantitative detection of SARS-CoV-2 RNA from the nasal wash of all the clinical samples was done by performing rtRT-PCR using Simplexa™ COVID-19 Direct kit (DiaSorin) as per the manufacturer's instructions. Reactions were run on the LIAISON® MDX instrument (DiaSorin). The DiaSorin Molecular Simplexa™ COVID-19 Direct assay system is a real-time RT-PCR system that enables the direct amplification of Coronavirus SARS-CoV-2 RNA from nasal wash (NW) specimen. The data is analyzed with LIAISON® MDX Studio Software. The assay targets two different regions of the SARS-CoV-2 genome, ORF1ab S gene 14 (link) . The sample was considered SARS-CoV-2 positive when one of the target genes in the same sample was positive and negative if both the genes were negative. The Ct values for both the genes were nearly similar. In this study, we used the Ct value of ORF1ab from nasal wash for the comparisons. The Ct value is inversely correlated with RNA copy number of the virus. Ct value < 40 was considered COVID-19 positive.

Sharma S., Thiriard A., Olislagers V., Lechien J.R., Jurion M.H., Delforge M.L., Marchant A, & Saussez S. (2024). Mucosal antibody response and SARS-CoV-2 shedding in patients with COVID-19 related olfactory dysfunction. Journal of medical virology, 96(1).

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COVID-19 Viral RNA Quantification

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The viral RNA present in the culture medium of the samples described above were quantified by Ct measurement with Simplexa™ COVID-19 Direct assay (DiaSorin, Vicenza, Italy), as previously described [29 (link)] according to the manufacturer’s instructions. Briefly, 50 μL of culture medium and 50 μL of Reaction Mix were added to their specific wells on a direct amplification disk, which was loaded onto the LIAISON^® MDX instrument (DiaSorin).

Alonzi T., Aiello A., Petrone L., Najafi Fard S., D’Eletto M., Falasca L., Nardacci R., Rossin F., Delogu G., Castilletti C., Capobianchi M.R., Ippolito G., Piacentini M, & Goletti D. (2021). Cysteamine with In Vitro Antiviral Activity and Immunomodulatory Effects Has the Potential to Be a Repurposing Drug Candidate for COVID-19 Therapy. Cells, 11(1), 52.

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