20 / 22
Page 43
Notes:
conferenceseries
.com
Volume 8, Issue 5 (Suppl)
J Blood Disord Transfus, an open access journal
ISSN:2155-9864
Hematology 2017
November 08-09, 2017
November 08-09, 2017 | Las Vegas, USA
11
th
International Conference on
Hematology & Hematological Oncology
Screening and identification of an aptamer to theHL-60 cell line of acute promyelocytic leukemia froma specific
human ssDNA library
Nabusige Jean Brenda Gesa
Makerere University, Uganda
A
PL is the most curable form of AML due to its sensitivity to ATRA, but challenges due to the threat of DIC at diagnosis and
drug toxicity of combination therapies during treatment, still remain. The aim of this study was to generate information on a
newly selected group of ssDNA aptamer candidates and build a potential aptamer library, for future use and reference in improving
diagnostic and therapeutic efficacy in APL management. This study involved the amplification of 7 highly specific ssDNA sequences
of 39-40bp each flanked by an 18bp primer sequence on both 5’ and 3’ ends. DNA was cloned into DH5
α
cells and ssDNA obtained by
affinity chromatography, purified and quantified by spectrophotometry. Cell binding affinity assays were conducted with APL HL-60
cells at room temperature, with incubation at 37
0
c. Results were quantified by spectrophotometry. Quality and yield of PCR amplified
DNA was dependent on concentration of plasmid template in the PCR mix and the number of cycles employed. It was realized that
DNA purification using Phenol-chloroform was most effective when plasmid templates used were freshly extracted and hadn’t been
subjected to prolonged storage. Of the 7 DNA sequences tested, sequences 135, 57 and 2 were observed to have the highest affinities
and 59, the lowest. Sequences 135 and 2, on statistical analysis demonstrated the highest affinities and are deemed ideal candidates for
further investigation in the development of effective aptamers and other tools for timely diagnosis and effective management of APL.
We suggested that further research be done.
Biography
Nabusige Jean Brenda Gesa is a dedicated scholar of Hematology and Oncology, passionate about healthcare advancement in Uganda and proficient in at least 3 languag-
es, including English and Mandarin Chinese. She received her Bachelors’ degree in Biomedical Laboratory Technology from Makerere University, Kampala and a Master of
Medicine in Clinical Laboratory Diagnostics at Beihua University, China (2017). She also acquired diagnostic laboratory experience at St. Raphael of St. Francis Hospital,
Nsambya in Kampala and Jilin Central Hospital, China. Her research interests include environmental pollutants in relation to hematological malignancies and hematological
cancer research; diagnostics and therapeutics.
jeangesa@yahoo.comNabusige Jean Brenda Gesa, J Blood Disord Transfus 2017, 8:5 (Suppl)
DOI: 10.4172/2155-9864-C1-028