QUANTITATIVE COMPARISON OF NASBA-ELISA AND RT-PCR-ELISA SENSITIVITIES FOR MEASUREMENT OF THE BCR-ABL GENES FUSION TRANSCRIPT IN CML PATIENTS

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NAZEMI ALI; SADEGHIZADEH MAJID; FOROUZANDEH MOGHADAM M.; JAVADI GH.R.; HASHEMI MEHRDAD

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Journal: MEDICAL SCIENCES JOURNAL OF ISLAMIC AZAD UNIVERSITY Year:2008 | Volume:18 | Issue:2 (52) Page(s): 67-74

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Title

QUANTITATIVE COMPARISON OF NASBA-ELISA AND RT-PCR-ELISA SENSITIVITIES FOR MEASUREMENT OF THE BCR-ABL GENES FUSION TRANSCRIPT IN CML PATIENTS

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Keywords

-PCR-ELISAQ3

NASBA-ELISAQ3

BCR/ABL TRANSCRIPTQ3

Abstract

Background: Chronic myeloid leukemia (CML) is characterized by neoplastic overproduction of myelocytes and neutrophils. Affected patients have a Philadelphia chromosome which arises following a translocation between long arms of chromosome 9 and 22 (q34;q11). This results in abelson murine/breakpoint cluster region (BCR/ABL) fusion. Detection of cells carrying BCR/ABL fusion is extremely important in monitoring response to treatment, remission and relapse in CML patients. In this study, we compared RT-PCR and NASBA techniques to determine quantitatively the number of BCR/ABL TRANSCRIPTs.Materials and Methods: Fusion transcripts were synthesized and RNA was extracted from K562 leukemic cell line. A serial dilution of both fusion transcript and RNA was prepared; then sensitivities of both techniques were determined. RT-PCR and NASBA reaction products were labeled using equal ratios of DIG-11-dUTP and DIG-11-UTP respectively. Following denaturation, hybridization reactions were carried out with specific probes. The products were incubated in streptavidin coated microplates. Then, the plates were washed, anti-DIG conjugated with peroxidase added and using ATBS as substrate, enzymatic activity was determined by absorption at 405nm.Results: The results showed that specificity of two techniques was equal but RT-PCR-ELISA sensitivity was about 100-fold more than NASBA-ELISA as it could detect 100pg RNA less than NASBA-ELISA (0.006 versus 0.06pg RNA). Furthermore, leukemia cell detection precision by RT-PCR-ELISA and NASBA-ELISA was 4 and 400 cells, respectively.Conclusion: While NASBA technique does not need thermal cycler PCR but has less sensitivity than RT-PCR and is not suitable for quantitative assessment.

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APA: Copy

NAZEMI, ALI, SADEGHIZADEH, MAJID, FOROUZANDEH MOGHADAM, M., JAVADI, GH.R., & HASHEMI, MEHRDAD. (2008). QUANTITATIVE COMPARISON OF NASBA-ELISA AND RT-PCR-ELISA SENSITIVITIES FOR MEASUREMENT OF THE BCR-ABL GENES FUSION TRANSCRIPT IN CML PATIENTS. MEDICAL SCIENCES JOURNAL OF ISLAMIC AZAD UNIVERSITY, 18(2 (52)), 67-74. SID. https://sid.ir/paper/98525/en

Vancouver: Copy

NAZEMI ALI, SADEGHIZADEH MAJID, FOROUZANDEH MOGHADAM M., JAVADI GH.R., HASHEMI MEHRDAD. QUANTITATIVE COMPARISON OF NASBA-ELISA AND RT-PCR-ELISA SENSITIVITIES FOR MEASUREMENT OF THE BCR-ABL GENES FUSION TRANSCRIPT IN CML PATIENTS. MEDICAL SCIENCES JOURNAL OF ISLAMIC AZAD UNIVERSITY[Internet]. 2008;18(2 (52)):67-74. Available from: https://sid.ir/paper/98525/en

IEEE: Copy

ALI NAZEMI, MAJID SADEGHIZADEH, M. FOROUZANDEH MOGHADAM, GH.R. JAVADI, and MEHRDAD HASHEMI, “QUANTITATIVE COMPARISON OF NASBA-ELISA AND RT-PCR-ELISA SENSITIVITIES FOR MEASUREMENT OF THE BCR-ABL GENES FUSION TRANSCRIPT IN CML PATIENTS,” MEDICAL SCIENCES JOURNAL OF ISLAMIC AZAD UNIVERSITY, vol. 18, no. 2 (52), pp. 67–74, 2008, [Online]. Available: https://sid.ir/paper/98525/en

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