A new real-time reverse transcription-PCR assay for detection of human enterovirus 68 in respiratory samples

doi:10.1128/JCM.03691-14

. 2015 May;53(5):1725-6.

doi: 10.1128/JCM.03691-14. Epub 2015 Feb 18.

A new real-time reverse transcription-PCR assay for detection of human enterovirus 68 in respiratory samples

Antonio Piralla¹, Alessia Girello¹, Marta Premoli¹, Fausto Baldanti²

Affiliations

¹ Molecular Virology Unit, Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
² Molecular Virology Unit, Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy Section of Microbiology, Department of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, Italy f.baldanti@smatteo.pv.it.

PMID: 25694533
PMCID: PMC4400749
DOI: 10.1128/JCM.03691-14

A new real-time reverse transcription-PCR assay for detection of human enterovirus 68 in respiratory samples

Antonio Piralla et al. J Clin Microbiol. 2015 May.

. 2015 May;53(5):1725-6.

doi: 10.1128/JCM.03691-14. Epub 2015 Feb 18.

Authors

Antonio Piralla¹, Alessia Girello¹, Marta Premoli¹, Fausto Baldanti²

Affiliations

¹ Molecular Virology Unit, Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
² Molecular Virology Unit, Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy Section of Microbiology, Department of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, Italy f.baldanti@smatteo.pv.it.

PMID: 25694533
PMCID: PMC4400749
DOI: 10.1128/JCM.03691-14

Abstract

A global reemergence of human enterovirus 68 (EV-D68) associated with severe respiratory illness occurred in 2014. We developed and validated an EV-D68-specific real-time reverse transcription-PCR (RT-PCR) for the detection of EV-D68 in respiratory samples. The rapid diagnosis of EV-D68 may contribute to better management of EV-D68 infections.

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Figures

**FIG 1**
(A) Linear range for the EV-D68 real-time RT-PCR assay. The standard curve was obtained by plotting the *C_T* values against the starting copy number. The coefficient R² of linear regression is shown. (B) Probit regression analysis to determine the limit of detection (LOD) for the EV-D68-specific real-time RT-PCR. TheLOD with a 95% probability is represented by a gray dotted line.

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Cited by

Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay.
Wylie TN, Wylie KM, Buller RS, Cannella M, Storch GA. Wylie TN, et al. J Clin Microbiol. 2015 Aug;53(8):2641-7. doi: 10.1128/JCM.00923-15. Epub 2015 Jun 10. J Clin Microbiol. 2015. PMID: 26063859 Free PMC article.
Emergence of divergent enterovirus (EV) D68 sub-clade D1 strains, northern Italy, September to October 2018.
Pellegrinelli L, Giardina F, Lunghi G, Uceda Renteria SC, Greco L, Fratini A, Galli C, Piralla A, Binda S, Pariani E, Baldanti F. Pellegrinelli L, et al. Euro Surveill. 2019 Feb;24(7):1900090. doi: 10.2807/1560-7917.ES.2018.24.7.1900090. Euro Surveill. 2019. PMID: 30782269 Free PMC article.
Enterovirus-D68 (EV-D68) in pediatric patients with respiratory infection: The circulation of a new B3 clade in Italy.
Piralla A, Principi N, Ruggiero L, Girello A, Giardina F, De Sando E, Caimmi S, Bianchini S, Marseglia GL, Lunghi G, Baldanti F, Esposito S. Piralla A, et al. J Clin Virol. 2018 Feb-Mar;99-100:91-96. doi: 10.1016/j.jcv.2018.01.005. Epub 2018 Jan 12. J Clin Virol. 2018. PMID: 29396353 Free PMC article.
Phylogenetic characteristics and molecular epidemiological analysis of novel enterovirus EV-B83 isolated from Tibet, China.
Xiao J, Zhang Y, Hong M, Han Z, Zhang M, Song Y, Yan D, Zhu S, Xu W. Xiao J, et al. Sci Rep. 2020 Apr 20;10(1):6630. doi: 10.1038/s41598-020-63691-4. Sci Rep. 2020. PMID: 32313119 Free PMC article.
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Ikuse T, Aizawa Y, Takihara H, Okuda S, Watanabe K, Saitoh A. Ikuse T, et al. J Clin Microbiol. 2021 Oct 19;59(11):e0115121. doi: 10.1128/JCM.01151-21. Epub 2021 Aug 25. J Clin Microbiol. 2021. PMID: 34432489 Free PMC article.

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References

1. Oberste MS, Maher K, Schnurr D, Flemister MR, Lovchik JC, Peters H, Sessions W, Kirk C, Chatterjee N, Fuller S, Hanauer JM, Pallansch MA. 2004. Enterovirus 68 is associated with respiratory illness and shares biological features with both the enteroviruses and the rhinoviruses. J Gen Virol 85:2577–2584. doi:10.1099/vir.0.79925-0. - DOI - PubMed
1. Centers for Disease Control and Prevention. 2011. Clusters of acute respiratory illness associated with human enterovirus 68—Asia, Europe, and United States, 2008–2010. MMWR Morb Mortal Wkly Rep 60:1301–1304. - PubMed
1. Meijer A, Benschop K, Donker G, van der Avoort H. 2014. Continued seasonal circulation of enterovirus D68 in the Netherlands, 2011–2014. Euro Surveill 19:20935 http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20935. - PubMed
1. Piralla A, Girello A, Grignani M, Gozalo-Margüello M, Marchi A, Marseglia G, Baldanti F. 2014. Phylogenetic characterization of enterovirus 68 strains in patients with respiratory syndromes in Italy. J Med Virol 86:1590–1593. doi:10.1002/jmv.23821. - DOI - PMC - PubMed
1. Piralla A, Lilleri D, Sarasini A, Marchi A, Zecca M, Stronati M, Baldanti F, Gerna G. 2012. Human rhinovirus and human respiratory enterovirus (EV68 and EV104) infections in hospitalized patients in Italy, 2008–2009. Diagn Microbiol Infect Dis 73:162–167. doi:10.1016/j.diagmicrobio.2012.02.019. - DOI - PubMed

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[1] Oberste MS, Maher K, Schnurr D, Flemister MR, Lovchik JC, Peters H, Sessions W, Kirk C, Chatterjee N, Fuller S, Hanauer JM, Pallansch MA. 2004. Enterovirus 68 is associated with respiratory illness and shares biological features with both the enteroviruses and the rhinoviruses. J Gen Virol 85:2577–2584. doi:10.1099/vir.0.79925-0. - DOI - PubMed

[2] Oberste MS, Maher K, Schnurr D, Flemister MR, Lovchik JC, Peters H, Sessions W, Kirk C, Chatterjee N, Fuller S, Hanauer JM, Pallansch MA. 2004. Enterovirus 68 is associated with respiratory illness and shares biological features with both the enteroviruses and the rhinoviruses. J Gen Virol 85:2577–2584. doi:10.1099/vir.0.79925-0. - DOI - PubMed

[3] Centers for Disease Control and Prevention. 2011. Clusters of acute respiratory illness associated with human enterovirus 68—Asia, Europe, and United States, 2008–2010. MMWR Morb Mortal Wkly Rep 60:1301–1304. - PubMed

[4] Centers for Disease Control and Prevention. 2011. Clusters of acute respiratory illness associated with human enterovirus 68—Asia, Europe, and United States, 2008–2010. MMWR Morb Mortal Wkly Rep 60:1301–1304. - PubMed

[5] Meijer A, Benschop K, Donker G, van der Avoort H. 2014. Continued seasonal circulation of enterovirus D68 in the Netherlands, 2011–2014. Euro Surveill 19:20935 http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20935. - PubMed

[6] Meijer A, Benschop K, Donker G, van der Avoort H. 2014. Continued seasonal circulation of enterovirus D68 in the Netherlands, 2011–2014. Euro Surveill 19:20935 http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20935. - PubMed

[7] Piralla A, Girello A, Grignani M, Gozalo-Margüello M, Marchi A, Marseglia G, Baldanti F. 2014. Phylogenetic characterization of enterovirus 68 strains in patients with respiratory syndromes in Italy. J Med Virol 86:1590–1593. doi:10.1002/jmv.23821. - DOI - PMC - PubMed

[8] Piralla A, Girello A, Grignani M, Gozalo-Margüello M, Marchi A, Marseglia G, Baldanti F. 2014. Phylogenetic characterization of enterovirus 68 strains in patients with respiratory syndromes in Italy. J Med Virol 86:1590–1593. doi:10.1002/jmv.23821. - DOI - PMC - PubMed

[9] Piralla A, Lilleri D, Sarasini A, Marchi A, Zecca M, Stronati M, Baldanti F, Gerna G. 2012. Human rhinovirus and human respiratory enterovirus (EV68 and EV104) infections in hospitalized patients in Italy, 2008–2009. Diagn Microbiol Infect Dis 73:162–167. doi:10.1016/j.diagmicrobio.2012.02.019. - DOI - PubMed

[10] Piralla A, Lilleri D, Sarasini A, Marchi A, Zecca M, Stronati M, Baldanti F, Gerna G. 2012. Human rhinovirus and human respiratory enterovirus (EV68 and EV104) infections in hospitalized patients in Italy, 2008–2009. Diagn Microbiol Infect Dis 73:162–167. doi:10.1016/j.diagmicrobio.2012.02.019. - DOI - PubMed

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A new real-time reverse transcription-PCR assay for detection of human enterovirus 68 in respiratory samples

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A new real-time reverse transcription-PCR assay for detection of human enterovirus 68 in respiratory samples

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