The Q-LAMP Method Represents a Valid and Rapid Alternative for the Detection of the BCR-ABL1 Rearrangement in Philadelphia-Positive Leukemias
Abstract
:1. Introduction
2. Results
2.1. Concordance between the Q-LAMP Assay and the Standard BIOMED-1 Method on p210 and p190 BCR-ABL1 Isoforms
2.2. Q-LAMP Performance on Common BCR-ABL1 p210 Isoforms
2.3. Concordance between the Q-LAMP Assay and the Standard BIOMED-1 Method on Rare p210 and p190 BCR-ABL1 Isoforms
3. Discussion
4. Materials and Methods
4.1. Patients
4.2. RNA Extraction
4.3. BCR-ABL1 Q-LAMP Assay
4.4. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ABL1 | Abelson murine leukemia |
ALL | acute lymphoblastic leukemia |
BCR | breakpoint cluster region |
CML | chronic myeloid leukemia |
GUSβ | β-glucuronidase |
LAMP | loop-mediated isothermal amplification |
ROC | receiver operating characteristic |
TK | tyrosine kinase |
TKI | tyrosine kinase inhibitor |
TFR | treatment-free remission |
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Results by Conventional RT-PCR (BIOMED-1) | |||||
---|---|---|---|---|---|
p190 | p210 | Negative | Total | ||
Q-LAMP Assay | p190 | 14 | / | / | 14 |
p210 | / | 108 | / | 107 | |
Negative | / | / | 50 | 50 | |
Total | 14 | 108 | 50 | 172 |
Isoform (N Sample) | Mean Tt * (st. dev) | Median Tt * (Range) | CV |
---|---|---|---|
e1a2 | 27.03 | 26.70 | 0.07 |
(14) | (1.87) | (24.45–31.80) | / |
e13a2 | 18.64 | 18.48 | 0.08 |
(44) | (1.41) | (15.25–21.67) | / |
e14a2 | 25.46 | 26.08 | 0.13 |
(43) | (3.29) | (16.53–34.57) | / |
e13a2/e14a2 | 19.84 | 19.63 | 0.06 |
(21) | (1.29) | (17.93–23.20) | / |
Results by conventional RT-PCR (BIOMED-1) | ||||||
---|---|---|---|---|---|---|
e1a3 | e13a3 (b2a3) | e14a2 (b3a3) | e19a2 | Total | ||
Q-LAMP Assay | e1a3 | 5 | / | / | / | 5 |
e13a2 (b2a3) | / | 9 | / | / | 9 | |
e14a2 (b3a3) | / | / | 3 | / | 3 | |
e19a2 | / | / | / | 3 | 3 | |
Total | 5 | 9 | 3 | 3 | 20 |
Q-LAMP Assay | BIOMED-1 Method | |
---|---|---|
Nucleic acid input (RNA ng/sample) | RNA (500) | cDNA (1000) |
Number of steps | 1 | ≥3 |
Reaction setup | Easy | Moderately complex (RT-PCR is required) |
Detection of rare BCR-ABL1 isoforms | Possible | Possible |
Time to results (minutes) | <60 | >300 |
Mark | CE/IVD | LDT |
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Stella, S.; Gottardi, E.M.; Favout, V.; Barragan Gonzalez, E.; Errichiello, S.; Vitale, S.R.; Fava, C.; Luciano, L.; Stagno, F.; Grimaldi, F.; et al. The Q-LAMP Method Represents a Valid and Rapid Alternative for the Detection of the BCR-ABL1 Rearrangement in Philadelphia-Positive Leukemias. Int. J. Mol. Sci. 2019, 20, 6106. https://doi.org/10.3390/ijms20246106
Stella S, Gottardi EM, Favout V, Barragan Gonzalez E, Errichiello S, Vitale SR, Fava C, Luciano L, Stagno F, Grimaldi F, et al. The Q-LAMP Method Represents a Valid and Rapid Alternative for the Detection of the BCR-ABL1 Rearrangement in Philadelphia-Positive Leukemias. International Journal of Molecular Sciences. 2019; 20(24):6106. https://doi.org/10.3390/ijms20246106
Chicago/Turabian StyleStella, Stefania, Enrico Marco Gottardi, Valeria Favout, Eva Barragan Gonzalez, Santa Errichiello, Silvia Rita Vitale, Carmen Fava, Luigia Luciano, Fabio Stagno, Francesco Grimaldi, and et al. 2019. "The Q-LAMP Method Represents a Valid and Rapid Alternative for the Detection of the BCR-ABL1 Rearrangement in Philadelphia-Positive Leukemias" International Journal of Molecular Sciences 20, no. 24: 6106. https://doi.org/10.3390/ijms20246106