Rapid visualization molecular fluorescence detection of methicillin‐resistant Staphylococcus aureus using the multiplex MIRA‐qPCR method

Abstract

Multidrug‐resistant (MDR) bacterial infections constitute a major public health problem worldwide. A rapid method for the detection of methicillin‐resistant Staphylococcus aureus (MRSA) is critical for the timely prevention of bacterial infections and the accurate clinical use of drugs. The nuc and mecA genes are potentially indicative of MRSA infection and in this study, a multiplex molecular fluorescence multi‐enzyme isothermal rapid amplification visual assay was proposed and established. The method is capable of detecting MRSA at 17 min, 40°C amplification, and is well differentiated from common clinical bacteria in specific assays, with 500 colony‐forming units/mL of MRSA detected under optimal conditions. This method has excellent diagnostic capabilities versus classical methods to detect clinical samples and shows potential in the identification of pathogenic microorganisms in a clinical setting.This article is protected by copyright. All rights reserved

Establishment and application of quantitative detection of Bacillus velezensis HMB26553, a biocontrol agent against cotton damping‐off caused by Rhizoctonia

“…Several powerful detection methods for identifying target microorganisms had been developed and were widely used, including culturedependent techniques with an antibiotic-resistant mutant, enzymelinked immunosorbent assay (ELISA), loop-mediated isothermal amplification (LAMP), and nucleic acid amplification-based methods like polymerase chain reaction (PCR) and quantitative PCR (qPCR). [4][5][6][7][8][9] Among them, qPCR has reliably detected and quantified species such as Plectosphaerella cucumerina, Metarhizium clade 1, Clonostachys rosea, [10][11][12] and specific strains such as Pantoea agglomerans strain CPA-2, Trichoderma asperellum strain icc012, Trichoderma gamsii strain icc080, Trichoderma atroviride strain SC1, Bacillus subtilis strain QST713, Bacillus amyloliquefaciens strain D747, Chaetomium globosum, Fusarium oxysporum strain Fo47, and Pseudomonas fluorescens strain EPS62e. [13][14][15][16][17][18][19][20] In genus Bacillus, many studies have focused on the specific detection and quantification of B. cereus group due to its human pathogenicity by qPCR, [21][22][23][24][25] but few have reported critical markers for the specific detection and quantification of other Bacillus species, [26][27][28] specifically for strain-specific detection in Bacillus velezensis.…”