The Use of Next-Generation Sequencing for the Quality Control of Live-Attenuated Polio Vaccines

Charlton, Hockley, Laassri, Wilton, Crawt, Preston, , Rigsby, Chumakov, Martin (2020) The Use of Next-Generation Sequencing for the Quality Control of Live-Attenuated Polio Vaccines J Infect Dis (IF: 6.4) 222(11) 1920-1927

Abstract

Next-generation sequencing (NGS) analysis was compared to the current MAPREC (mutational analysis by polymerase chain reaction and restriction enzyme cleavage) assay for quality control of live-attenuated oral polio vaccine (OPV).MAPREC measures reversion of the main OPV attenuating mutations such as uracil (U) to cytosine (C) at nucleotide 472 in the 5' noncoding region of type 3 OPV. Eleven type 3 OPV samples were analyzed by 8 laboratories using their in-house NGS method.Intraassay, intralaboratory, and interlaboratory variability of NGS 472-C estimates across samples and laboratories were very low, leading to excellent agreement between laboratories. A high degree of correlation between %472-C results by MAPREC and NGS was observed in all laboratories (Pearson correlation coefficient r = 0.996). NGS estimates of sequences at nucleotide 2493 with known polymorphism among type 3 OPV lots also produced low assay variability and excellent between-laboratory agreement.The high consistency of NGS data demonstrates that NGS analysis can be used as high-resolution test alternative to MAPREC, producing whole-genome profiles to evaluate OPV production consistency, possibly eliminating the need for tests in animals. This would be very beneficial for the quality assessment of next-generation polio vaccines and, eventually, for other live-attenuated viral vaccines.© Crown copyright 2020.

Links

http://www.ncbi.nlm.nih.gov/pubmed/32492703
http://dx.doi.org/10.1093/infdis/jiaa299

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