Quantification of the middle east respiratory syndrome-coronavirus rna in tissues by quantitative real-time RT-PCR

Abdullah Algaissi, Anurodh S. Agrawal, Anwar M. Hashem, Chien Te K. Tseng

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Since the emergence of the Middle East respiratory syndrome-coronavirus (MERS-CoV) in 2012, more than 2280 confirmed human infections and 800 associated deaths had been reported to the World Health Organization. MERS-CoV is a single-stranded RNA virus that belongs to the Coronaviridae family. MERS-CoV infection leads to a variety of clinical outcomes in humans ranging from asymptomatic and mild infection to severe acute lung injury and multi-organ failure and death. To study the pathogenesis of MERS-CoV infection and development of medical countermeasures (MCMs) for MERS, a number of genetically modified mouse models have been developed, including various versions of transgenic mice expressing the human DPP4 viral receptor. Tracking and quantifying viral infection, among others, in permissive hosts is a key endpoint for studying MERS pathogenesis and evaluating the efficacy of selected MCMs developed for MERS. In addition to quantifying infectious progeny virus which requires high-containment biosafety level (BSL)-3 laboratory, here we outlined an established real-time quantitative RT-PCR (RT-qPCR)-based procedure to unequivocally quantify MERS-CoV-specific RNAs within the lungs of infected human DPP4 (hDPP4, transgenic (hDPP4 Tg) mice under a standard BSL-2 laboratory.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages99-106
Number of pages8
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2099
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

RNA
Coronavirus Infections
Transgenic Mice
Coronaviridae
Asymptomatic Infections
Acute Lung Injury
RNA Viruses
Virus Diseases
Middle East Respiratory Syndrome Coronavirus
Viruses
Lung
Infection

Keywords

  • Animal models
  • MERS-CoV
  • RT-qPCR

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Algaissi, A., Agrawal, A. S., Hashem, A. M., & Tseng, C. T. K. (2020). Quantification of the middle east respiratory syndrome-coronavirus rna in tissues by quantitative real-time RT-PCR. In Methods in Molecular Biology (pp. 99-106). (Methods in Molecular Biology; Vol. 2099). Humana Press Inc.. https://doi.org/10.1007/978-1-0716-0211-9_8

Quantification of the middle east respiratory syndrome-coronavirus rna in tissues by quantitative real-time RT-PCR. / Algaissi, Abdullah; Agrawal, Anurodh S.; Hashem, Anwar M.; Tseng, Chien Te K.

Methods in Molecular Biology. Humana Press Inc., 2020. p. 99-106 (Methods in Molecular Biology; Vol. 2099).

Research output: Chapter in Book/Report/Conference proceedingChapter

Algaissi, A, Agrawal, AS, Hashem, AM & Tseng, CTK 2020, Quantification of the middle east respiratory syndrome-coronavirus rna in tissues by quantitative real-time RT-PCR. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2099, Humana Press Inc., pp. 99-106. https://doi.org/10.1007/978-1-0716-0211-9_8
Algaissi A, Agrawal AS, Hashem AM, Tseng CTK. Quantification of the middle east respiratory syndrome-coronavirus rna in tissues by quantitative real-time RT-PCR. In Methods in Molecular Biology. Humana Press Inc. 2020. p. 99-106. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-0716-0211-9_8
Algaissi, Abdullah ; Agrawal, Anurodh S. ; Hashem, Anwar M. ; Tseng, Chien Te K. / Quantification of the middle east respiratory syndrome-coronavirus rna in tissues by quantitative real-time RT-PCR. Methods in Molecular Biology. Humana Press Inc., 2020. pp. 99-106 (Methods in Molecular Biology).
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