Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines
Background/Objectives: Conventional live oral poliovirus vaccines (OPVs) effectively prevent poliomyelitis. These vaccines are derived from three attenuated Sabin strains of poliovirus, which can revert within the first week of replication to a neurovirulent phenotype, leading to sporadic cases of v...
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MDPI AG
2025-01-01
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| author | Olga Singh Hasmik Manukyan Erman Tritama Shwu-Maan Lee Jerry P. Weir Majid Laassri |
| author_facet | Olga Singh Hasmik Manukyan Erman Tritama Shwu-Maan Lee Jerry P. Weir Majid Laassri |
| author_sort | Olga Singh |
| collection | DOAJ |
| description | Background/Objectives: Conventional live oral poliovirus vaccines (OPVs) effectively prevent poliomyelitis. These vaccines are derived from three attenuated Sabin strains of poliovirus, which can revert within the first week of replication to a neurovirulent phenotype, leading to sporadic cases of vaccine-associated paralytic poliomyelitis (VAPP) among vaccinees and their contacts. A novel OPV2 vaccine (nOPV2) with enhanced genetic stability was developed recently; type 1 and type 3 nOPV strains were engineered using the nOPV2 genome as a backbone by replacing the capsid precursor polyprotein (P1) with that of Sabin strains type 1 and type 3, respectively. The nOPV vaccines have a high degree of sequence homology with the parental Sabin 2 genome, and some manufacturing facilities produce and store both Sabin OPV and nOPV. Therefore, detecting Sabin virus contaminations in nOPV lots is crucial. Methods: This study describes the development of pan quantitative reverse transcription polymerase chain reaction (panRT-PCR) and multiplex one-step RT-PCR (mosRT-PCR) assays for the straightforward detection and identification of contaminating Sabin viruses when present in significantly higher amounts of nOPV strains. Results: The two assays exhibit high specificity, reproducibility, and sensitivity to detect 0.0001% and 0.00001% of Sabin viruses in nOPV, respectively. Additionally, an analysis of 12 trivalent nOPV formulation lots using both methods confirmed that the nOPV lots were free from Sabin virus contamination. Conclusions: The results demonstrated that the RT-PCR assays are sensitive and specific. These assays are relevant for quality control and lot release of nOPV vaccines. |
| format | Article |
| id | doaj-art-8e20c745336d4ea69f1ab51a037b87d4 |
| institution | Kabale University |
| issn | 2076-393X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj-art-8e20c745336d4ea69f1ab51a037b87d42025-01-24T13:51:51ZengMDPI AGVaccines2076-393X2025-01-011317510.3390/vaccines13010075Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus VaccinesOlga Singh0Hasmik Manukyan1Erman Tritama2Shwu-Maan Lee3Jerry P. Weir4Majid Laassri5Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USACenter for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USAResearch and Development Division, PT. BioFarma, Bandung, West Java 40161, IndonesiaCenter for Vaccine Innovation and Access, PATH, Seattle, WA 98121, USACenter for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USACenter for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USABackground/Objectives: Conventional live oral poliovirus vaccines (OPVs) effectively prevent poliomyelitis. These vaccines are derived from three attenuated Sabin strains of poliovirus, which can revert within the first week of replication to a neurovirulent phenotype, leading to sporadic cases of vaccine-associated paralytic poliomyelitis (VAPP) among vaccinees and their contacts. A novel OPV2 vaccine (nOPV2) with enhanced genetic stability was developed recently; type 1 and type 3 nOPV strains were engineered using the nOPV2 genome as a backbone by replacing the capsid precursor polyprotein (P1) with that of Sabin strains type 1 and type 3, respectively. The nOPV vaccines have a high degree of sequence homology with the parental Sabin 2 genome, and some manufacturing facilities produce and store both Sabin OPV and nOPV. Therefore, detecting Sabin virus contaminations in nOPV lots is crucial. Methods: This study describes the development of pan quantitative reverse transcription polymerase chain reaction (panRT-PCR) and multiplex one-step RT-PCR (mosRT-PCR) assays for the straightforward detection and identification of contaminating Sabin viruses when present in significantly higher amounts of nOPV strains. Results: The two assays exhibit high specificity, reproducibility, and sensitivity to detect 0.0001% and 0.00001% of Sabin viruses in nOPV, respectively. Additionally, an analysis of 12 trivalent nOPV formulation lots using both methods confirmed that the nOPV lots were free from Sabin virus contamination. Conclusions: The results demonstrated that the RT-PCR assays are sensitive and specific. These assays are relevant for quality control and lot release of nOPV vaccines.https://www.mdpi.com/2076-393X/13/1/75viral contaminationOPVnOPVRT-PCRvaccinespoliovirus |
| spellingShingle | Olga Singh Hasmik Manukyan Erman Tritama Shwu-Maan Lee Jerry P. Weir Majid Laassri Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines Vaccines viral contamination OPV nOPV RT-PCR vaccines poliovirus |
| title | Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines |
| title_full | Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines |
| title_fullStr | Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines |
| title_full_unstemmed | Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines |
| title_short | Development of RT-PCR Assays for Simple Detection and Identification of Sabin Virus Contaminants in the Novel Oral Poliovirus Vaccines |
| title_sort | development of rt pcr assays for simple detection and identification of sabin virus contaminants in the novel oral poliovirus vaccines |
| topic | viral contamination OPV nOPV RT-PCR vaccines poliovirus |
| url | https://www.mdpi.com/2076-393X/13/1/75 |
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