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dc.contributor.authorOnwuamah, Chika K
dc.contributor.authorOkwuraiwe, Azuka P
dc.contributor.authorAhmed, Rahaman A
dc.contributor.authorSokei, Judith O
dc.contributor.authorPonmak, Jamda
dc.contributor.authorOkoli, Leona C
dc.contributor.authorKagurusi, Brian A
dc.contributor.authorAnejo-Okopi, Joseph
dc.date.accessioned2020-12-08T19:39:31Z
dc.date.available2020-12-08T19:39:31Z
dc.date.issued2020-12-31
dc.identifier.urihttp://hdl.handle.net/10713/14217
dc.description.abstractDespite various challenges that hinder the implementation of high-tech molecular methods in resource-limited settings, we have been able to implement and achieve International Organization for Standardization 15189:2012 accreditation for genotypic HIV drug resistance testing in our facility. At the Center for Human Virology and Genomics, Nigerian Institute of Medical Research, Nigeria has recorded a high sequencing success rate and good quality sequence data. This was achieved by optimizing laboratory processes from 2008 to the current date. We have optimized sample preparation, RT-PCR, several post-PCR processes, and the cycle sequencing to improve the sensitivity of amplification even with limited plasma samples and low viral copy numbers. The optimized workflow maximizes output, minimizes reagent wastage, and achieves substantial cost savings without compromising the quality of the sequence data. Our performance at our last external quality assurance program is a testimonial to the efficiency of the workflow. For the 5-sample panel, each with 67–68 mutation points evaluated, we scored 100% for all 5 specimens. Our optimized laboratory workflow is thus documented to support laboratories and to help researchers achieve excellent results the first time and eliminate contamination while minimizing the wastage of costly sequencing reagents.en_US
dc.description.urihttps://doi.org/10.7171/jbt.20-3104-006en_US
dc.language.isoenen_US
dc.publisherAssociation of Biomolecular Resource Facilitiesen_US
dc.relation.ispartofJournal of Biomolecular Techniques : JBTen_US
dc.rights© Association of Biomolecular Resource Facilities.en_US
dc.subjectbiomedical researchen_US
dc.subjectHIV drug resistanceen_US
dc.subjectlaboratory workflowen_US
dc.subjectreverse transcriptase polymerase chain reactionen_US
dc.titleLaboratory Optimization Tweaks for Sanger Sequencing in a Resource-Limited Settingen_US
dc.typeArticleen_US
dc.identifier.doi10.7171/jbt.20-3104-006
dc.identifier.pmid33100921
dc.source.volume31
dc.source.issue4
dc.source.beginpage157
dc.source.endpage164
dc.source.countryUnited States


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