Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10157909 | Molecular and Cellular Probes | 2018 | 27 Pages |
Abstract
Streptococcus pyogenes (Group A Streptococcus, GAS) and Streptococcus agalactiae (Group B Streptococcus, GBS) are common pathogens that threaten public health. In this study, a double recombinase polymerase (RPA) amplification assay was developed to rapidly detect these pathogens. Specificity tests revealed that the GAS and GBS strains were positive for speB and SIP genes, respectively. In clinical samples, the double assay performed similarly to the traditional biochemical method. The limits of detection were both â¤100 copies per reaction. In tests for simulant-contaminated samples, bacterial-culture media containing 103â¯CFU/mL original concentrations of S. pyogenes and S. agalactiae were positive in RPA assays after incubating for 4â¯h. Results can be obtained at 37â¯Â°C in 20â¯min. To determine whether propidium monoazide (PMA) can eliminate the influence of DNA extracted from dead cells, a bacterial suspension was treated with PMA before DNA extraction. Findings of RPA assay showed that DNA extracted from dead cells had no fluorescence signal. Therefore, the PMA-RPA assay is a promising technology for field tests and rapid point-of-care diagnosis.
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Authors
Jing Chen, Yuanyang Wang, Xiaoqing Liu, Guopei Chen, Xuejian Chen, Jiaping Chen, Zhongdong Liu, Jingwen Gong, Guowu Yang, Quanxue Lan,