Duplex real-time fluorescent recombinase polymerase amplification for the rapid and sensitive detection of two resistance genes in drug-resistant Staphylococcus aureus

doi:10.1016/j.mimet.2022.106590

	Duplex real-time fluorescent recombinase polymerase amplification for the rapid and sensitive detection of two resistance genes in drug-resistant Staphylococcus aureus
	Shi, Zhonglin 1,3; Li, Yanan 3,4; Hu, Anzhong 3; Cui, Junsheng 3; Shao, Min 2; Zhu, Ling 3; Yang, Ke 3; Liu, Yong 3; Deng, Guoqing 3; Zhu, Cancan 3
刊名	JOURNAL OF MICROBIOLOGICAL METHODS
	2022-11-01
卷号	202
关键词	Drug -resistant Staphylococcus aureus ermA mecA Recombinase polymerase amplification Resistance genes
ISSN号	0167-7012
DOI	10.1016/j.mimet.2022.106590
通讯作者	Zhu, Cancan(zhucancan@aiofm.ac.cn)
英文摘要	In the clinic, drug-resistant Staphylococcus aureus (S. aureus) is the most common suppurative infection pathogen in humans. It can cause local infections in humans and animals, such as pneumonia, mastitis, and other systemic illnesses. At present, the detection of drug-resistant S. aureus includes traditional isolation by culture and anti-microbial susceptibility tests. However, these methods are complicated in experimental design, specialized in operation and time consuming. Therefore, a rapid and accurate drug-resistant S. aureus detection technology is urgently needed. In this study, we combined duplex pairs of fluorescent probes with recombinase polymerase amplification (RPA) to realize the simultaneous detection of two resistance genes in drug-resistant S. aureus. The method shows low detection limit, detecting 20 copies within 10 min. The analytical specificity of this method was evaluated with several related drug-resistant bacterial strains (Non-resistant S. aureus, Acinetobacter bau-mannii, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae), and the positive signal was only observed with drug-resistant S. aureus. In addition, the clinical suitability of this method was verified by 30 clinical isolates. Compared with qPCR, the coincidence rate of drug resistance genes were 100% (mecA) and 96.7% (ermA), respectively. These results show that the duplex real-time fluorescent RPA assay is a rapid, low detection limit and specific detection of mecA and ermA genes in isolates of drug-resistant S. aureus.
资助项目	Natural Science Foundation of Anhui Province of China[82002189] ; Key Research and Development Program of Anhui Province of China[2008085QF311] ; Youth Innovation Promotion Association of the Chinese Academy of Sciences[202004d7020014] ; Youth Innovation Promotion Association of the Chinese Academy of Sciences[2018489] ; Dean's Fund of Hefei Institute of Physical Science, China[2021442] ; National Natural Science Foundation of China[YZJJ2020QN35]
WOS关键词	ISOTHERMAL AMPLIFICATION ; CHAIN-REACTION ; ASSAY ; PATHOGENS ; POINT ; DNA
WOS研究方向	Biochemistry & Molecular Biology ; Microbiology
语种	英语
出版者	ELSEVIER
WOS记录号	WOS:000877555600008
资助机构	Natural Science Foundation of Anhui Province of China ; Key Research and Development Program of Anhui Province of China ; Youth Innovation Promotion Association of the Chinese Academy of Sciences ; Dean's Fund of Hefei Institute of Physical Science, China ; National Natural Science Foundation of China
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/130307]
专题	中国科学院合肥物质科学研究院
通讯作者	Zhu, Cancan
作者单位	1.Anhui Univ, Inst Phys Sci & Informat Technol, Hefei 230601, Anhui, Peoples R China 2.Anhui Med Univ, Affiliated Hosp 1, Dept Crit Care Med, Hefei 230031, Anhui, Peoples R China 3.Chinese Acad Sci, Anhui Inst Opt & Fine Mech, Hefei Inst Phys Sci, Hefei 230031, Anhui, Peoples R China 4.Univ Sci & Technol China, Hefei 230026, Anhui, Peoples R China
推荐引用方式 GB/T 7714	Shi, Zhonglin,Li, Yanan,Hu, Anzhong,et al. Duplex real-time fluorescent recombinase polymerase amplification for the rapid and sensitive detection of two resistance genes in drug-resistant Staphylococcus aureus[J]. JOURNAL OF MICROBIOLOGICAL METHODS,2022,202.
APA	Shi, Zhonglin.,Li, Yanan.,Hu, Anzhong.,Cui, Junsheng.,Shao, Min.,...&Zhu, Cancan.(2022).Duplex real-time fluorescent recombinase polymerase amplification for the rapid and sensitive detection of two resistance genes in drug-resistant Staphylococcus aureus.JOURNAL OF MICROBIOLOGICAL METHODS,202.
MLA	Shi, Zhonglin,et al."Duplex real-time fluorescent recombinase polymerase amplification for the rapid and sensitive detection of two resistance genes in drug-resistant Staphylococcus aureus".JOURNAL OF MICROBIOLOGICAL METHODS 202(2022).