A real-time PCR assay for detection of emerging infectious Elizabethkingia miricola

Qi Zhang; Ruixue Hu; Zemao Gu

doi:10.1016/j.mcp.2020.101571

A real-time PCR assay for detection of emerging infectious Elizabethkingia miricola

Mol Cell Probes. 2020 Aug:52:101571. doi: 10.1016/j.mcp.2020.101571. Epub 2020 Apr 11.

Authors

Qi Zhang¹, Ruixue Hu¹, Zemao Gu²

Affiliations

¹ Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China; Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention, Wuhan, 430070, China; ShuangshuishuiShuanglü Institute, Huazhong Agricultural University, Wuhan, 430070, China.
² Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China; Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention, Wuhan, 430070, China; ShuangshuishuiShuanglü Institute, Huazhong Agricultural University, Wuhan, 430070, China. Electronic address: guzemao@mail.hzau.edu.cn.

PMID: 32289377
DOI: 10.1016/j.mcp.2020.101571

Abstract

Elizabethkingia miricola, a Gram-negative bacillus, is emerging as a life-threatening pathogen in both humans and animals. However, no specific and rapid diagnostic method exists to detect E. miricola. Here, we established a real-time PCR assay for the rapid, sensitive, and specific detection of E. miricola with a wide dynamic range of 10⁸ copies/μL to 10² copies/μL. The detection limit of the real-time assay was 145 copies/μL, which was 100 times more sensitive than conventional PCR. All clinical isolates E. miricola from different host species yield very close Tm (80.25 ± 0.25 °C). Additionally, no cross-reaction or false positives were observed in the assay for non-target bacterial species. The performance of this assay was primarily assessed by testing frog tissue samples. Overall, our study provided a real-time PCR assay, which is a rapid, sensitive, and specific diagnostic method that could be used for early diagnosis and epidemiological investigation of E. miricola.

Keywords: Elizabethkingia miricola; Real-time PCR; Specific detection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Anura / microbiology
Flavobacteriaceae / genetics*
Flavobacteriaceae / isolation & purification*
Real-Time Polymerase Chain Reaction / methods*
Reference Standards
Reproducibility of Results
Sensitivity and Specificity

Supplementary concepts

Elizabethkingia miricola