title: Evaluation of in-house genotyping assay performance using dried blood spot specimens in the Global World Health Organization laboratory network.
abstract: In resource-limited settings, there is increased demand for human immunodeficiency virus type 1 drug resistance testing. Because preservation of plasma specimens is often not feasible in resource-limited settings, use of dried blood spots (DBSs) is being adopted. We used 2 panels of DBSs for genotyping assay validation and proficiency testing in selected laboratories in the World Health Organization laboratory network in 14 countries. An amplification sensitivity of 1000 copies/mL was achieved by 2 laboratories. Reproducibility and accuracy of nucleotide sequence determination and resistance-associated mutation identification from DBSs was similar to that previously determined for plasma. International shipping at ambient temperature had no significant effect on amplification success. These studies indicate that DBS-based genotyping is equally reproducible and reliable, although slightly less sensitive, compared with plasma.
authors.affiliation: Data First Consulting, Belmont, California 94002, USA. nparkin34@gmail.com
authors.forename: Neil
authors.initials: N
authors.lastname: Parkin
authors.forename: Carmen
authors.initials: C
authors.lastname: de Mendoza
authors.forename: Rob
authors.initials: R
authors.lastname: Schuurman
authors.forename: Cheryl
authors.initials: C
authors.lastname: Jennings
authors.forename: James
authors.initials: J
authors.lastname: Bremer
authors.forename: Michael R
authors.initials: MR
authors.lastname: Jordan
authors.forename: Silvia
authors.initials: S
authors.lastname: Bertagnolio
mesh_terms: D059788:Dried Blood Spot Testing / Q000379:methods* / Q000592:standards; D024882:Drug Resistance, Viral / Q000235:genetics; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods* / Q000592:standards; D015658:HIV Infections / Q000097:blood* / Q000821:virology*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007753:Laboratories; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013696:Temperature; D019562:Viral Load / Q000379:methods / Q000592:standards; D014944:World Health Organization