579. Epidemiological Data Differences between Gel-purified vs. DNA-purified Oral Polio Vaccine in environmental samples

Abstract Background As wild poliovirus is eradicated, preventing circulation of vaccine-derived top priority. Our lab developed real-time multiplex PCR assays and deep sequencing methodology to detect characterize OPV strains from stool samples. The method requires gel purification product created viral RNA in However, the process filters out a significant portion Here, we compare gel- vs. DNA- sample retainment for downstream analysis. Methods 554 samples qPCR positive at least one serotype previous study were used. There are 268 1 (S1) isolates, 405 2 (S2) 318 3 (S3) isolates. amplicons ran through 0.08% agarose identity presence ∼3.5kb amplicon interest. then underwent either gel-purification spin column kit or DNA kit. Samples with concentration >10ng/uL elution required NGS. Purification Flowchart Graph 1:Gel-purification, DNA-purification, suggested workflow charts Results Comparing 168 S2 DNA-purified, 90 bands identified both protocols, 100% DNA-purified 35.6% gel-purified had >10ng/ul. 41.3% banded < =34CT >10ng/ul Of 43 pediatric participants, prior IPV did not impact proportion number individuals viable NGS (16/31 (51.6%) vs 25/43 (58.1%) respectively). Among processed via purification, median per participant dropped (IQR=1-3) (IQR=1-1). From same set samples, reduced vaccinated children 38 (82.6%), their household contacts (4.3%), community 6 (13%) 21 (80.7%) children, (3.8%) contacts, 4 (15.3%) contacts. Table 1:Purification outcome unique-participant-day (UPD) DNA-purificationGraph 2:Demographic information patient loss form on 2a. Sample collection across different days pre 32 UPD post gel-purification. 2b. does spread each unique-participant (UP) 25 UP 2c. Vaccination status (OPV child, contact, unvaccinated contact) 90UPD 2d. Number collected fron 3:Demographic 75 31 24 41 75UPD Conclusion Conclusions: Gel-purification causes reduction numbers variation, causing an incomplete data misrepresentation analysis, particularly within host variation may be most optimal Analysis all isolates process. Disclosures Yvonne A. Maldonado, MD, Pfizer: Grant/Research Support|Pfizer: Member, DSMB, Pfizer Meningococcal Vaccine clinical trial.