In-vitro culture of Plasmodium falciparum: Utility of modified (RPNI) medium for drug-sensitivity studies using SYBR Green I assay |
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Authors: | Shubhra Singh Rajeev Kumar Srivastava Mukesh Srivastava SK Puri K Srivastava |
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Affiliation: | aParasitology Division, Central Drug Research Institute, CSIR, Chhatar Manzil Palace, Post Box No. 173, Lucknow-226001, India;bBiometry & Statistics Division, Central Drug Research Institute, CSIR, Chhatar Manzil Palace, Post Box No. 173, Lucknow-226001, India |
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Abstract: | Studies were carried out to establish the potential of RPNI medium for drug-sensitivity studies using the MSF assay. The drug sensitivity of standard anti-malarials was compared using both the (3H) Hypoxanthine incorporation assay and the MSF assay. The media supplements used during the study have been human serum, FBS and ALBUMAX-II. Drug sensitivity of two parasite lines, adapted to grow separately in conventional as well as in RPNI medium was compared to observe the effect of RPNI medium on functional characteristics of the parasite. The results revealed identical IC50 values of standard anti-malarials obtained by both the (3H) Hypoxanthine incorporation assay and the MSF assay and no untoward effect of FBS and ALBUMAX-II could be noticed on the chemo-sensitivity of standard anti-malarials. Apart from this the chemo-sensitive response of parasite line adapted to grow in RPNI medium was observed to be intact. These findings showed that RPNI medium has potential to be used for chemo-sensitivity studies and the MSF assay being more convenient was observed to be most suitable assay for bio evaluation of new molecules. |
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Keywords: | (3H) Hypoxanthine Chloroquine Mefloquine Artemether DHA |
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