1. The Use of Next-Generation Sequencing for the Quality Control of Live-Attenuated Polio Vaccines.
- Author
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Charlton B, Hockley J, Laassri M, Wilton T, Crawt L, Preston M, Rigsby P, Chumakov K, and Martin J
- Subjects
- Animals, Humans, Mutation, Poliovirus, Polymerase Chain Reaction methods, High-Throughput Nucleotide Sequencing methods, Poliomyelitis prevention & control, Poliovirus Vaccine, Oral standards, Quality Control, Vaccines, Attenuated standards
- Abstract
Background: 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)., Methods: 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., Results: 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., Conclusions: 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.)
- Published
- 2020
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