A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic
Molecular testing of the <i>BCR-ABL1</i> transcript via real-time quantitative-polymerase-chain-reaction is the most sensitive approach for monitoring the response to tyrosine-kinase-inhibitors therapy in chronic myeloid leukaemia (CML) patients. Each stage of the molecular procedure has...
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doaj-605f9d39700947e9b08dab2fb0be37e42021-08-26T13:40:39ZengMDPI AGDiagnostics2075-44182021-08-01111502150210.3390/diagnostics11081502A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 PandemicStefania Stella0Silvia Rita Vitale1Michele Massimino2Adriana Puma3Cristina Tomarchio4Maria Stella Pennisi5Elena Tirrò6Chiara Romano7Federica Martorana8Fabio Stagno9Francesco Di Raimondo10Livia Manzella11Department of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyCenter of Experimental Oncology and Hematology, A.O.U. Policlinico “G.Rodolico—San Marco”, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyDivision of Hematology and Bone Marrow Transplant, A.O.U. Policlinico “G.Rodolico—San Marco”, 95123 Catania, ItalyDivision of Hematology and Bone Marrow Transplant, A.O.U. Policlinico “G.Rodolico—San Marco”, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Catania, 95123 Catania, ItalyMolecular testing of the <i>BCR-ABL1</i> transcript via real-time quantitative-polymerase-chain-reaction is the most sensitive approach for monitoring the response to tyrosine-kinase-inhibitors therapy in chronic myeloid leukaemia (CML) patients. Each stage of the molecular procedure has been standardized and optimized, including the total white blood cells (WBCs) and RNA isolation methods. Here, we compare the performance of our current manual protocol to a newly semiautomatic method based on the Biomek i-5 Automated Workstations integrated with the CytoFLEX Flow Cytometer, followed by the automatic QIAsymphony system to facilitate high-throughput processing samples and reduce the hands-on time and the risk associated with SARS-CoV-2. The recovery efficiency was investigated in blood samples from 100 adults with CML. We observe a 100% of concordance between the two methods, with similar total WBCs isolated (median 1.137 × 10<sup>6</sup> for manual method vs. 1.076 × 10<sup>6</sup> for semiautomatic system) and a comparable quality and quantity of RNA extracted (median 103 ng/μL with manual isolation kit vs. 99.95 ng/μL with the QIAsymphony system). Moreover, by stratifying patients according to their <i>BCR-ABL1</i> transcript levels, we obtained similar <i>BCR-ABL1/ABL1<sup>IS</sup></i> values and <i>ABL1</i> copies, and matched samples were assigned to the same group of molecular response. We conclude that this newly semiautomatic workflow has a performance comparable to our more laborious standard manual, which can be replaced, particularly when specimens from patients with suspected or confirmed SARS-CoV-2 infection need to be processed.https://www.mdpi.com/2075-4418/11/8/1502chronic myeloid leukaemiamolecular responseQ-PCRSARS-CoV-2 infection<i>BCR-ABL1/ABL1<sup>IS</sup></i> |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Stefania Stella Silvia Rita Vitale Michele Massimino Adriana Puma Cristina Tomarchio Maria Stella Pennisi Elena Tirrò Chiara Romano Federica Martorana Fabio Stagno Francesco Di Raimondo Livia Manzella |
spellingShingle |
Stefania Stella Silvia Rita Vitale Michele Massimino Adriana Puma Cristina Tomarchio Maria Stella Pennisi Elena Tirrò Chiara Romano Federica Martorana Fabio Stagno Francesco Di Raimondo Livia Manzella A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic Diagnostics chronic myeloid leukaemia molecular response Q-PCR SARS-CoV-2 infection <i>BCR-ABL1/ABL1<sup>IS</sup></i> |
author_facet |
Stefania Stella Silvia Rita Vitale Michele Massimino Adriana Puma Cristina Tomarchio Maria Stella Pennisi Elena Tirrò Chiara Romano Federica Martorana Fabio Stagno Francesco Di Raimondo Livia Manzella |
author_sort |
Stefania Stella |
title |
A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic |
title_short |
A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic |
title_full |
A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic |
title_fullStr |
A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic |
title_full_unstemmed |
A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic |
title_sort |
novel system for semiautomatic sample processing in chronic myeloid leukaemia: increasing throughput without impacting on molecular monitoring at time of sars-cov-2 pandemic |
publisher |
MDPI AG |
series |
Diagnostics |
issn |
2075-4418 |
publishDate |
2021-08-01 |
description |
Molecular testing of the <i>BCR-ABL1</i> transcript via real-time quantitative-polymerase-chain-reaction is the most sensitive approach for monitoring the response to tyrosine-kinase-inhibitors therapy in chronic myeloid leukaemia (CML) patients. Each stage of the molecular procedure has been standardized and optimized, including the total white blood cells (WBCs) and RNA isolation methods. Here, we compare the performance of our current manual protocol to a newly semiautomatic method based on the Biomek i-5 Automated Workstations integrated with the CytoFLEX Flow Cytometer, followed by the automatic QIAsymphony system to facilitate high-throughput processing samples and reduce the hands-on time and the risk associated with SARS-CoV-2. The recovery efficiency was investigated in blood samples from 100 adults with CML. We observe a 100% of concordance between the two methods, with similar total WBCs isolated (median 1.137 × 10<sup>6</sup> for manual method vs. 1.076 × 10<sup>6</sup> for semiautomatic system) and a comparable quality and quantity of RNA extracted (median 103 ng/μL with manual isolation kit vs. 99.95 ng/μL with the QIAsymphony system). Moreover, by stratifying patients according to their <i>BCR-ABL1</i> transcript levels, we obtained similar <i>BCR-ABL1/ABL1<sup>IS</sup></i> values and <i>ABL1</i> copies, and matched samples were assigned to the same group of molecular response. We conclude that this newly semiautomatic workflow has a performance comparable to our more laborious standard manual, which can be replaced, particularly when specimens from patients with suspected or confirmed SARS-CoV-2 infection need to be processed. |
topic |
chronic myeloid leukaemia molecular response Q-PCR SARS-CoV-2 infection <i>BCR-ABL1/ABL1<sup>IS</sup></i> |
url |
https://www.mdpi.com/2075-4418/11/8/1502 |
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