Optimized RNA extraction for detection of BCR-ABL transcripts in chronic myeloid leukemia patients during treatment

Choo Yuen Ting, Norazrina Azmi, Mohd Makmor Bakry, Ahmad Fuad Shamsuddin

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Objective: Monitoring of treatment response in chronic myeloid leukemia (CML) is difficult in leukopenia patients due to poor quality of blood cell RNA sample. Plasma RNA serves as an alternative for quantification of mutation gene but reproducibility depends on the quality and quantity of extracted RNA. The present study aimed to evaluate eight commercially available RNA extraction kits for detection of BCR-ABL transcripts in CML patients during treatment. Methods: RNA extraction was carried out using eight types of commercially available kits on plasma samples of chronic myeloid leukemia patients on tyrosine kinase inhibitors therapy. Plasma samples of healthy individuals were used as controls. Results: GeneAll® RiboEx™ with minor protocol modification demonstrated pure and sufficient amount of plasma RNA for quantification of ABL control gene and BCR-ABL mutation gene. This protocol produced a mean of 13.67±0.65 ng/μL plasma RNA whereas the purity indicator A260/280 ratio was 1.96±0.16 and A260/230 ratio was 2.07±0.12. As for gene expression analysis, 3561±392 copies of ABL control gene and 8.9±1.9 copies of BCR-ABL mutation gene were successfully amplified from 150 ng of plasma RNA. Conclusion: GeneAll® RiboEx™ with minor protocol modification produced purer plasma RNA extracts compared to other tested kits.

Original languageEnglish
Pages (from-to)415-418
Number of pages4
JournalInternational Journal of Pharmacy and Pharmaceutical Sciences
Volume6
Issue number1
Publication statusPublished - 2014

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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
RNA
Therapeutics
Genes
Mutation
Leukopenia
Protein-Tyrosine Kinases
Blood Cells
Gene Expression

Keywords

  • Chronic myeloid leukemia
  • Peripheral blood
  • Plasma RNA
  • RNA extraction

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Pharmacology

Cite this

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title = "Optimized RNA extraction for detection of BCR-ABL transcripts in chronic myeloid leukemia patients during treatment",
abstract = "Objective: Monitoring of treatment response in chronic myeloid leukemia (CML) is difficult in leukopenia patients due to poor quality of blood cell RNA sample. Plasma RNA serves as an alternative for quantification of mutation gene but reproducibility depends on the quality and quantity of extracted RNA. The present study aimed to evaluate eight commercially available RNA extraction kits for detection of BCR-ABL transcripts in CML patients during treatment. Methods: RNA extraction was carried out using eight types of commercially available kits on plasma samples of chronic myeloid leukemia patients on tyrosine kinase inhibitors therapy. Plasma samples of healthy individuals were used as controls. Results: GeneAll{\circledR} RiboEx™ with minor protocol modification demonstrated pure and sufficient amount of plasma RNA for quantification of ABL control gene and BCR-ABL mutation gene. This protocol produced a mean of 13.67±0.65 ng/μL plasma RNA whereas the purity indicator A260/280 ratio was 1.96±0.16 and A260/230 ratio was 2.07±0.12. As for gene expression analysis, 3561±392 copies of ABL control gene and 8.9±1.9 copies of BCR-ABL mutation gene were successfully amplified from 150 ng of plasma RNA. Conclusion: GeneAll{\circledR} RiboEx™ with minor protocol modification produced purer plasma RNA extracts compared to other tested kits.",
keywords = "Chronic myeloid leukemia, Peripheral blood, Plasma RNA, RNA extraction",
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TY - JOUR

T1 - Optimized RNA extraction for detection of BCR-ABL transcripts in chronic myeloid leukemia patients during treatment

AU - Ting, Choo Yuen

AU - Azmi, Norazrina

AU - Makmor Bakry, Mohd

AU - Shamsuddin, Ahmad Fuad

PY - 2014

Y1 - 2014

N2 - Objective: Monitoring of treatment response in chronic myeloid leukemia (CML) is difficult in leukopenia patients due to poor quality of blood cell RNA sample. Plasma RNA serves as an alternative for quantification of mutation gene but reproducibility depends on the quality and quantity of extracted RNA. The present study aimed to evaluate eight commercially available RNA extraction kits for detection of BCR-ABL transcripts in CML patients during treatment. Methods: RNA extraction was carried out using eight types of commercially available kits on plasma samples of chronic myeloid leukemia patients on tyrosine kinase inhibitors therapy. Plasma samples of healthy individuals were used as controls. Results: GeneAll® RiboEx™ with minor protocol modification demonstrated pure and sufficient amount of plasma RNA for quantification of ABL control gene and BCR-ABL mutation gene. This protocol produced a mean of 13.67±0.65 ng/μL plasma RNA whereas the purity indicator A260/280 ratio was 1.96±0.16 and A260/230 ratio was 2.07±0.12. As for gene expression analysis, 3561±392 copies of ABL control gene and 8.9±1.9 copies of BCR-ABL mutation gene were successfully amplified from 150 ng of plasma RNA. Conclusion: GeneAll® RiboEx™ with minor protocol modification produced purer plasma RNA extracts compared to other tested kits.

AB - Objective: Monitoring of treatment response in chronic myeloid leukemia (CML) is difficult in leukopenia patients due to poor quality of blood cell RNA sample. Plasma RNA serves as an alternative for quantification of mutation gene but reproducibility depends on the quality and quantity of extracted RNA. The present study aimed to evaluate eight commercially available RNA extraction kits for detection of BCR-ABL transcripts in CML patients during treatment. Methods: RNA extraction was carried out using eight types of commercially available kits on plasma samples of chronic myeloid leukemia patients on tyrosine kinase inhibitors therapy. Plasma samples of healthy individuals were used as controls. Results: GeneAll® RiboEx™ with minor protocol modification demonstrated pure and sufficient amount of plasma RNA for quantification of ABL control gene and BCR-ABL mutation gene. This protocol produced a mean of 13.67±0.65 ng/μL plasma RNA whereas the purity indicator A260/280 ratio was 1.96±0.16 and A260/230 ratio was 2.07±0.12. As for gene expression analysis, 3561±392 copies of ABL control gene and 8.9±1.9 copies of BCR-ABL mutation gene were successfully amplified from 150 ng of plasma RNA. Conclusion: GeneAll® RiboEx™ with minor protocol modification produced purer plasma RNA extracts compared to other tested kits.

KW - Chronic myeloid leukemia

KW - Peripheral blood

KW - Plasma RNA

KW - RNA extraction

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