Quantitative multiplex one-step RT-PCR assay for identification and quantitation of Sabin strains of poliovirus in clinical and environmental specimens

Manukyan, Hasmik; Zagorodnyaya, Tatiana; Rüttimann, Ricardo; Manor, Yossi; Bandyopadhyay, Ananda S; Shulman, Lester M.; Chumakov, Konstantin; Laassri, Majid

doi:10.1016/j.jviromet.2018.06.009

Cited by 15 publications

(27 citation statements)

References 19 publications

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“…However, the current standard method of environmental surveillance of polio is the tissue-culture-based intratypic differentiation (ITD) assay, which is slower than direct qPCR and not quantitative (52,53). Although presence/absence data are valuable, we should move to quantitative assays as we approach eradication.…”

Section: Discussionmentioning

confidence: 99%

Epidemiology of the silent polio outbreak in Rahat, Israel, based on modeling of environmental surveillance data

Brouwer

Eisenberg

Pomeroy

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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139

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Israel experienced an outbreak of wild poliovirus type 1 (WPV1) in 2013–2014, detected through environmental surveillance of the sewage system. No cases of acute flaccid paralysis were reported, and the epidemic subsided after a bivalent oral polio vaccination (bOPV) campaign. As we approach global eradication, polio will increasingly be detected only through environmental surveillance. We developed a framework to convert quantitative polymerase chain reaction (qPCR) cycle threshold data into scaled WPV1 and OPV1 concentrations for inference within a deterministic, compartmental infectious disease transmission model. We used this approach to estimate the epidemic curve and transmission dynamics, as well as assess alternate vaccination scenarios. Our analysis estimates the outbreak peaked in late June, much earlier than previous estimates derived from analysis of stool samples, although the exact epidemic trajectory remains uncertain. We estimate the basic reproduction number was 1.62 (95% CI 1.04–2.02). Model estimates indicate that 59% (95% CI 9–77%) of susceptible individuals (primarily children under 10 years old) were infected with WPV1 over a little more than six months, mostly before the vaccination campaign onset, and that the vaccination campaign averted 10% (95% CI 1–24%) of WPV1 infections. As we approach global polio eradication, environmental monitoring with qPCR can be used as a highly sensitive method to enhance disease surveillance. Our analytic approach brings public health relevance to environmental data that, if systematically collected, can guide eradication efforts.

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Section: Discussionmentioning

confidence: 99%

Epidemiology of the silent polio outbreak in Rahat, Israel, based on modeling of environmental surveillance data

Brouwer

Eisenberg

Pomeroy

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

139

120

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“…Virus titer was determined by 50% end-point dilution assay (50% cell culture infectious dose, TCID50) using HEp-2 cells (ATCC CCL-23), a HeLa sub-line (ATCC, VA, USA). Viral RNA copy numbers in the culture fluid was also measured by real-time reverse transcription-PCR 20,21 . Supplementary Fig.…”

Section: Construction Of a Vector For The Crispr/cas9 System Guide Smentioning

confidence: 99%

Poliovirus-nonsusceptible Vero cell line for the World Health Organization global action plan

Okemoto-Nakamura

Someya

Yamaji

et al. 2020

Preprint

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Polio or poliomyelitis is a disabling and life-threatening disease caused by poliovirus (PV). As a consequence of global polio vaccination efforts, wild PV serotype 2 has been eradicated, and wild PV serotypes 1- and 3-transmitted cases have been largely eliminated except for in limited regions around the world. However, vaccine-derived PV, pathogenically reverted live PV vaccine strains in vaccinated humans, has become a serious issue. For the global eradication of polio, the World Health Organization is conducting the third edition of the Global Action Plan, which is requesting stringent control of potentially PV-infected materials. To facilitate the mission, we generated a PV-nonsusceptible Vero cell subline, which may serve as an ideal replacement of standard Vero cells to isolate emerging/re-emerging viruses without the risk of generating PV-infected materials.

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“…The quantitative multiplex one-step RT-PCR (qmosRT-PCR) was described previously [18]. Briefly, the qmosRT-PCR reactions were prepared in 96-well optical plates in a final volume of 25 µL using 2 µL of 1/20 diluted cell lysate and QuantiFast Multiplex RT-PCR Kit (QIAGEN, Valencia, CA, USA).…”

Section: Quantitative Multiplex One-step Rt-pcrmentioning

confidence: 99%

“…The three poliovirus serotypes were quantified in the same reaction of qmosRT-PCR as described previously [18] and summarized above. The positive and negative results for viral replication were recorded for each well and used for sera titers calculation for each poliovirus serotype in accordance with the Spearman-Karber formula [20].…”

Section: Mpbn Assay and Data Analysismentioning

confidence: 99%

“…This communication describes the development of a multiplex PCR-based neutralization (MPBN) assay for anti-poliovirus antibodies titration that uses a quantitative multiplex one-step RT-PCR (qmosRT-PCR) [18] as a read-out instead of CPE used in the conventional assay. Recently, we successfully used a similar approach to develop a multiplex PCR-based titration (MPBT) assay for the simultaneous titration of the three poliovirus serotypes [19].…”

Section: Introductionmentioning

confidence: 99%

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Multiplex PCR-Based Neutralization (MPBN) Assay for Titers Determination of the Three Types of Anti-Poliovirus Neutralizing-Antibodies

et al. 2020

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Determination of poliovirus-neutralizing antibodies is an important part of clinical studies of poliovirus vaccines, epidemiological surveillance and seroprevalence studies that are crucial for global polio eradication campaigns. The conventional neutralization test is based on inhibition of cytopathic effect caused by poliovirus by serial dilutions of test serum. It is laborious, time-consuming and not suitable for large scale analysis. To overcome these limitations, a multiplex PCR-based neutralization (MPBN) assay was developed to measure the neutralizing antibody titers of anti-poliovirus sera against three serotypes of the virus in the same reaction and in shorter time. All three anti-poliovirus sera types were analyzed in a single assay. The MPBN assay was reproducible, robust and sensitive. Its lower limits of titration for the three anti-poliovirus sera types were within range of 0.76–1.64 per mL. Different anti-poliovirus sera were tested with conventional and MPBN assays; the results obtained by both methods correlated well and generated similar results. The MPBN is the first neutralization assay that specifically titrates anti-poliovirus antibodies against the three serotypes of the virus in the same reaction; it can be completed in two to three days instead of ten days for the conventional assay and can be automated for high-throughput implementation.

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Quantitative multiplex one-step RT-PCR assay for identification and quantitation of Sabin strains of poliovirus in clinical and environmental specimens

Cited by 15 publications

References 19 publications

Epidemiology of the silent polio outbreak in Rahat, Israel, based on modeling of environmental surveillance data

Epidemiology of the silent polio outbreak in Rahat, Israel, based on modeling of environmental surveillance data

Poliovirus-nonsusceptible Vero cell line for the World Health Organization global action plan

Multiplex PCR-Based Neutralization (MPBN) Assay for Titers Determination of the Three Types of Anti-Poliovirus Neutralizing-Antibodies

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