Tags

Type your tag names separated by a space and hit enter

Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus.
J Clin Microbiol. 2014 Jan; 52(1):67-75.JC

Abstract

A new human coronavirus (CoV), subsequently named Middle East respiratory syndrome (MERS)-CoV, was first reported in Saudi Arabia in September 2012. In response, we developed two real-time reverse transcription-PCR (rRT-PCR) assays targeting the MERS-CoV nucleocapsid (N) gene and evaluated these assays as a panel with a previously published assay targeting the region upstream of the MERS-CoV envelope gene (upE) for the detection and confirmation of MERS-CoV infection. All assays detected ≤10 copies/reaction of quantified RNA transcripts, with a linear dynamic range of 8 log units and 1.3 × 10(-3) 50% tissue culture infective doses (TCID50)/ml of cultured MERS-CoV per reaction. All assays performed comparably with respiratory, serum, and stool specimens spiked with cultured virus. No false-positive amplifications were obtained with other human coronaviruses or common respiratory viral pathogens or with 336 diverse clinical specimens from non-MERS-CoV cases; specimens from two confirmed MERS-CoV cases were positive with all assay signatures. In June 2012, the U.S. Food and Drug Administration authorized emergency use of the rRT-PCR assay panel as an in vitro diagnostic test for MERS-CoV. A kit consisting of the three assay signatures and a positive control was assembled and distributed to public health laboratories in the United States and internationally to support MERS-CoV surveillance and public health responses.

Authors+Show Affiliations

Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

24153118

Citation

Lu, Xiaoyan, et al. "Real-time Reverse transcription-PCR Assay Panel for Middle East Respiratory Syndrome Coronavirus." Journal of Clinical Microbiology, vol. 52, no. 1, 2014, pp. 67-75.
Lu X, Whitaker B, Sakthivel SK, et al. Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus. J Clin Microbiol. 2014;52(1):67-75.
Lu, X., Whitaker, B., Sakthivel, S. K., Kamili, S., Rose, L. E., Lowe, L., Mohareb, E., Elassal, E. M., Al-sanouri, T., Haddadin, A., & Erdman, D. D. (2014). Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus. Journal of Clinical Microbiology, 52(1), 67-75. https://doi.org/10.1128/JCM.02533-13
Lu X, et al. Real-time Reverse transcription-PCR Assay Panel for Middle East Respiratory Syndrome Coronavirus. J Clin Microbiol. 2014;52(1):67-75. PubMed PMID: 24153118.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus. AU - Lu,Xiaoyan, AU - Whitaker,Brett, AU - Sakthivel,Senthil Kumar K, AU - Kamili,Shifaq, AU - Rose,Laura E, AU - Lowe,Luis, AU - Mohareb,Emad, AU - Elassal,Emad M, AU - Al-sanouri,Tarek, AU - Haddadin,Aktham, AU - Erdman,Dean D, Y1 - 2013/10/23/ PY - 2013/10/25/entrez PY - 2013/10/25/pubmed PY - 2014/7/26/medline SP - 67 EP - 75 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 52 IS - 1 N2 - A new human coronavirus (CoV), subsequently named Middle East respiratory syndrome (MERS)-CoV, was first reported in Saudi Arabia in September 2012. In response, we developed two real-time reverse transcription-PCR (rRT-PCR) assays targeting the MERS-CoV nucleocapsid (N) gene and evaluated these assays as a panel with a previously published assay targeting the region upstream of the MERS-CoV envelope gene (upE) for the detection and confirmation of MERS-CoV infection. All assays detected ≤10 copies/reaction of quantified RNA transcripts, with a linear dynamic range of 8 log units and 1.3 × 10(-3) 50% tissue culture infective doses (TCID50)/ml of cultured MERS-CoV per reaction. All assays performed comparably with respiratory, serum, and stool specimens spiked with cultured virus. No false-positive amplifications were obtained with other human coronaviruses or common respiratory viral pathogens or with 336 diverse clinical specimens from non-MERS-CoV cases; specimens from two confirmed MERS-CoV cases were positive with all assay signatures. In June 2012, the U.S. Food and Drug Administration authorized emergency use of the rRT-PCR assay panel as an in vitro diagnostic test for MERS-CoV. A kit consisting of the three assay signatures and a positive control was assembled and distributed to public health laboratories in the United States and internationally to support MERS-CoV surveillance and public health responses. SN - 1098-660X UR - https://www.unboundmedicine.com/medline/citation/24153118/Real_time_reverse_transcription_PCR_assay_panel_for_Middle_East_respiratory_syndrome_coronavirus_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=24153118 DB - PRIME DP - Unbound Medicine ER -