Development of a Quantitative One-Step RT-PCR Method for the Detection of Sabin 2 Virus Contamination in a Novel Oral Poliovirus Vaccine Type 2

Figures

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  Figure 1. Nucleotide sequences of domain V region of nOPV2 and Sabin 2 viruses. Genomic location of primers and TaqMan probe used for detection and quantitation of Sabin 2 virus in nOPV2 vaccine and mutation 481A→G are indicated.

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  Figure 2. The three repeats of qosRT-PCR analysis (each repeat is presented in one line) of RNA diluted samples: RNA was extracted from Sabin 2 virus (with titer of 8.34 Log10 CCID50/mL), serially diluted 10-times using 10-fold steps and subjected to qosRT-PCR analysis. The R2 is around 1 for each repeat of the analyzed samples.

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  Figure 3. Specific Sabin 2 virus identification with qosRT-PCR demonstrated by using Sabin 2 RNA and plasmid containing Sabin 2 genome (positive controls), and plasmid containing the nOPV2-c1genome and water (negative controls).

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  Figure 4. The three repeats of qosRT-PCR analysis (each repeat is presented in one line) of Sabin 2 virus in presence of prevalent amounts of nOPV2: Sabin 2 virus (titer 8.34 log10 CCID50/mL) was serially diluted in 10-fold steps in nOPV2c1-295 virus (titer of 8.07 log10 CCID50/mL). All diluted samples were subjected to RNA extraction and then to the assay analysis. The R2 is around 0.99 for each repeat of the analyzed samples.