Simultaneous Dual‐Gene Test of Methicillin‐Resistant Staphylococcus Aureus using an Argonaute‐Centered Portable and Visual Biosensor

Author:

Kou Jun1,Li Yaru1,Zhao Zhiying1,Qiao Jiali1,Zhang Qiang2,Han Xiao1,Cheng Xinkuan1,Man Shuli1,Ma Long1ORCID

Affiliation:

1. State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial Microbiology Ministry of Education Tianjin Key Laboratory of Industry Microbiology National and Local United Engineering Lab of Metabolic Control Fermentation Technology China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry College of Biotechnology Tianjin University of Science & Technology Tianjin 300457 China

2. Branch of Tianjin Third Central Hospital Tianjin 300250 China

Abstract

AbstractThe development of novel method for drug‐resistant bacteria detection is imperative. A simultaneous dual‐gene Test of methicillin‐resistant Staphylococcus aureus (MRSA) is developed using an Argonaute‐centered portable biosensor (STAR). This is the first report concerning Argonaute‐based pathogenic bacteria detection. Simply, the species‐specific mecA and nuc gene are isothermally amplified using loop‐mediated isothermal amplification (LAMP) technique, followed by Argonaute‐based detection enabled by its programmable, guided, sequence‐specific recognition and cleavage. With the strategy, the targeted nucleic acid signals gene are dexterously converted into fluorescent signals. STAR is capable of detecting the nuc gene and mecA gene simultaneously in a single reaction. The limit of detection is 10 CFU/mL with a dynamic range from 10 to 107 CFU/mL. The sample‐to‐result time is <65 min. This method is successfully adapted to detect clinical samples, contaminated foods, and MRSA‐infected animals. This work broadens the reach of Argonaute‐based biosensing and presents a novel bacterial point‐of‐need (PON) detection platform.

Funder

National Natural Science Foundation of China

Tianjin Municipal Science and Technology Committee

State Key Laboratory of Food Nutrition and Safety

Publisher

Wiley

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