Lu Wang,Jing Sun,Jiyu Zhao,Jieyu Bai,Yueling Zhang,Yao Zhu,Wanjiang Zhang,Chunlai Wang,Paul R Langford,Siguo Liu,Gang Li

Talanta.2023 Sep 16:267:125202.doi: 10.1016/j.talanta.2023.125202. Online ahead of print.

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.

Keywords: CRISPR; Cas12a; Diagnostics; Streptococcus suis serotype 2; crRNA.