Alignment of Qx100/Qx200 Droplet Digital (Bio-Rad) and QuantStudio 3D (Thermofisher) Digital PCR for Quantification of BCR-ABL1 in Ph+ Chronic Myeloid Leukemia

• Main Authors: Carmen Fava, Simona Bernardi, Enrico Marco Gottardi, Roberta Lorenzatti, Laura Galeotti, Francesco Ceccherini, Francesco Cordoni, Filomena Daraio, Emilia Giugliano, Aleksandar Jovanovski, Jessica Petiti, Marta Varotto, Davide Barberio, Giovanna Rege-Cambrin, Paola Berchialla, Veronica Sciannameo, Michele Malagola, Giuseppe Saglio, Domenico Russo
• Format: Article
• Language: English
• Published: MDPI AG 2021-05-01
• Series: Diseases
• Subjects: digital polymerase chain reaction; digital PCR; dPCR; chronic myeloid leukemia; CML; minimal residual disease
• Online Access: https://www.mdpi.com/2079-9721/9/2/35

In recent years, the digital polymerase chain reaction has received increasing interest as it has emerged as a tool to provide more sensitive and accurate detection of minimal residual disease. In order to start the process of data alignment, we assessed the consistency of the BCR-ABL1 quantification results of the analysis of 16 RNA samples at different levels of disease. The results were obtained by two different laboratories that relied on The Qx100/Qx200 Droplet Digital PCR System (Bio-Rad) and Quant Studio 3D dPCR System (Thermofisher) platforms. We assessed the compatibility between the estimated values by linear regression, Bland–Altman bias-plot, and Mann–Whitney nonparametric test. The results confirmed the compatibility of the measures, allowing us tocompute an ‘alignment factor’ (AF), equal to 1.41, which was further validated by a different series of experiments. We conclude that the performed measurements by the two laboratories are comparable, and also equalized through the introduction of an alignment factor.