(P30) HIV-1 drug resistance genotyping from dried blood spots using the Free Virus Elution (FVE) Technique, in Zimbabwe
Tafadzwa Dylan Nyakubaya
(1)Midlands State University (2)University of Zimbabwe College of Health Science
Optimisation of HIV-1 drug resistance genotyping from dried blood spots using the
free virus elution technique, in Zimbabwe.
Tafadzwa Dylan Nyakubaya 1 , Morleen Muteveri 1 , Vinie Kouamou 2 , Bhavni Varyani 3 and David Katzenstein 4 .
Midlands State University, Zimbabwe Department of Applied Biosciences and Biotechnology 1 . University of
Zimbabwe 2 . Biomedical Research and Training Institute 3 . Stanford University 4 .
Introduction:Dried blood spots (DBS) are an alternative specimen type for HIV drug resistance
genotyping in resource-limited settings. A simple and cheap extraction technique is needed to
facilitate genotyping testing from DBS for patients on antiretroviral treatment (ART) for
effective management of HIV infection and Acquired Immune Deficiency Syndrome (AIDS).
Method: A cross sectional study was conducted to determine the sensitivity of the free virus
elution technique for DBS with a viral load above and below 100 000 copies/ml to determine
the presence of drug resistance mutations within patients on ART. A total of 10 DBS were
obtained from patients on ART. Five samples had a viral load above 100,000 copies/ml and the
other five below 100,000 copies/ml across different ranges. Total nucleic acids were first
extracted using the FVE technique followed by HIV-1 RNA extraction. The extracted RNA was
reverse transcribed and amplified by the RT-PCR method. The 1.3kb generated amplicon was
then sequenced using the sanger sequencing method and drug resistance mutations were
analysed using the Stanford HIV drug resistance database.
Results: All the five DBS specimens (50%) that had a VL>100,000copies/ml were successfully
genotyped. The other five, which had VL 100 000copies/ml. A more sensitive and
specific method is needed to genotype from DBS in resource limited settings.
Tafadzwa Dylan Nyakubaya has completed his BSc in December2017 at the age of 23 years from
Midlands State University,Zimbabwe. He’s a graduate that is keen to develop the international
community through vibrant research and collaborative effort.
Presenting author detailsTafadzwa Dylan Nyakubaya