Evaluation of Multiplex Polymerase Chain Reaction (PCR) as Prognostic Tools for Chronic Myeloid Leukemia (CML) Patients in Resource-Limited Setting at Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia.

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Date

2021-09

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Addis Abeba University

Abstract

Background:-The hallmark diagnostic tool for Chronic myeloid leukemia (CML) is Philadelphia chromosome t (9; 22) (q34; q11)) which gives rise to bcr-abl1 fusion oncogenic protein. QrtPCR of bcr-abl1has been established as a prognostic tool since the introduction of TKI drugs. But, in a resource-limited setting, like Ethiopia, multiplex reverse transcriptase PCR, which is used to be as a screening tool, could be adopted into the clinic as a relative prognostic tool than relying on hematological response. Objective: - to evaluate multiplex polymerase chain reaction (PCR) as prognostic tools for chronic myeloid leukemia (CML) patients in resource-limited settings at Tikur Anbessa Specialized Hospital (TASH). Method: - Hospital-based Cross-sectional study design was used for a total of 114 confirmed CML patients who were enrolled at the Hematology Clinic of TASH. Multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) and quantitative reverse transcriptase PCR were performed at AHRI molecular lab, and Leipzig hematology clinic, respectively. The descriptive statistic and ROC curve using excel were used to assess the demographic feature of the patients and to evaluate the prognostic value, respectively, and thereby the most valuable cutoff multiplex PCR was calculated in comparison with QRT- PCR. Result: - of the total enrolled study subjects (114), most of the transcripts were major fusion gene (M-bcr-abl1), b2a2 (28.1%), b3a2 (49.1%), and co-expressed were b2a2/b3a2 (0.9%). All new cases and follow-up patients who were positive with the multiplex RT-PCR method for bcrabl transcript were more than 95% agreement with quantitative PCR compared to the limit of detection sated for multiplex PCR. Multiplex RT- PCR at a cut-off 0.31% IS and above, in comparison to the standard prognostic tools (Qrt PCR) has a comparable result; sensitivity 96.6% and specificity 88% at the area under the curve 0.977(97.7 %) and CI 95% (0.952 to 1, P-value 0.0001). Conclusion: - Follow-up of patients on the hematological parameter is rather crude and not sensitive enough to modify the treatment regimen. But, the use of multiplex PCR, besides as screening tools, can provide an early sign of relapse as low as 0.31% IS bcr-abl transcript. As compared to Qrt-PCR, it is relatively cheap and accessible to be adopted into the routine clinic.

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Keywords

BCR-ABL- RT-PCR, chronic myeloid leukemia, molecular response, Ethiopia.

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