Middle East respiratory coronavirus (MERS-CoV) spike (S) protein vesicular stomatitis virus pseudoparticle neutralization assays offer a reliable alternative to the conventional neutralization assay in human seroepidemiological studies

Sandra Lester; Jennifer Harcourt; Michael Whitt; Hail M Al-Abdely; Claire M Midgley; Abdulrahim M Alkhamis; Hani A Aziz Jokhdar; Abdullah M Assiri; Azaibi Tamin; Natalie Thornburg

doi:10.1099/acmi.0.000057

Middle East respiratory coronavirus (MERS-CoV) spike (S) protein vesicular stomatitis virus pseudoparticle neutralization assays offer a reliable alternative to the conventional neutralization assay in human seroepidemiological studies

Access Microbiol. 2019 Sep 11;1(9):e000057. doi: 10.1099/acmi.0.000057. eCollection 2019.

Authors

Affiliations

¹ Synergy America, Inc., Duluth, GA, USA.
² National Center for Immunization and Respiratory Diseases, Division of Viral Diseases, Respiratory Viruses Laboratory Branch, Centers for Disease Control and Prevention (CDC), Atlanta, Georgia, USA.
³ The University of Tennessee Health Science Center, Microbiology, Immunology, and Biochemistry, Memphis, TN, USA.
⁴ Ministry of Health, Riyadh, Saudi Arabia.

Abstract

Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel zoonotic coronavirus that was identified in 2012. MERS-CoV infection in humans can result in an acute, severe respiratory disease and in some cases multi-organ failure; the global mortality rate is approximately 35 %. The MERS-CoV spike (S) protein is a major target for neutralizing antibodies in infected patients. The MERS-CoV microneutralization test (MNt) is the gold standard method for demonstrating prior infection. However, this method requires the use of live MERS-CoV in biosafety level 3 (BSL-3) containment. The present work describes the generation and validation of S protein-bearing vesicular stomatitis virus (VSV) pseudotype particles (VSV-MERS-CoV-S) in which the VSV glycoprotein G gene has been replaced by the luciferase reporter gene, followed by the establishment of a pseudoparticle-based neutralization test to detect MERS-CoV neutralizing antibodies under BSL-2 conditions. Using a panel of human sera from confirmed MERS-CoV patients, the VSV-MERS-CoV particle neutralization assay produced results that were highly comparable to those of the microneutralization test using live MERS-CoV. The results suggest that the VSV-MERS-CoV-S pseudotype neutralization assay offers a highly specific, sensitive and safer alternative method to detect MERS-CoV neutralizing antibodies in human sera.

Keywords: Middle East respiratory syndrome coronavirus (MERS-CoV); luciferase; microneutralization test (MNt); neutralizing antibodies; pseudoparticle; vesicular stomatitis virus.