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Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
Published on: December 23, 2022
A CRISPR-Cas12a-based platform facilitates the detection and serotyping of Streptococcus suis serotype 2
Lu Wang1, Jing Sun2, Jiyu Zhao3
1State Key Laboratory for Animal Disease Control and Prevention, Division of Bacterial Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150069, China; College of Veterinary Medicine, Xinjiang Agricultural University, Xinjiang, 830052, China.
Abstract:
Streptococcus suis serotype 2 is an economically important zoonotic pathogen that causes septicemia, arthritis, and meningitis in pigs and humans. S. suis serotype 2 is responsible for substantial economic losses to the swine industry and poses a serious threat to public health, and accurate and rapid detection is important for the prevention and control of epidemic disease. In this study, we developed a high-fidelity detection and serotyping platform for S. suis serotype 2 based on recombinase polymerase amplification (RPA) and a clustered regularly interspaced short palindromic repeat (CRISPR)-Cas12a system called Cards-SSJ/K. Cards-SSJ had a detection limit of 10 CFU, takes <60 min, and no cross-reaction was found with other S. suis serotypes, closely related Streptococcus spp., or common pig pathogens, and Cards-SSK could differentiate serotype 2 from serotype 1/2. Results from Cards-SSJ and qPCR were equivalent in detecting S. suis serotype 2 in tissue samples. Analysis indicated that despite a relatively high reagent cost compared to PCR and qPCR, Cards-SSJ was less time-consuming and had low requirements for equipment and personnel. Thus, it is an excellent method for point-of-care detection for S. suis serotype 2.
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