A CRISPR-Cas12a-based platform facilitates the detection and serotyping of Streptococcus suis serotype 2

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

doi:10.1016/j.talanta.2023.125202

A CRISPR-Cas12a-based platform facilitates the detection and serotyping of Streptococcus suis serotype 2

Talanta. 2024 Jan 15:267:125202. doi: 10.1016/j.talanta.2023.125202. Epub 2023 Sep 16.

Authors

Lu Wang¹, Jing Sun², Jiyu Zhao³, Jieyu Bai¹, Yueling Zhang², Yao Zhu², Wanjiang Zhang², Chunlai Wang², Paul R Langford⁴, Siguo Liu⁵, Gang Li⁶

Affiliations

¹ State 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.
² State Key Laboratory for Animal Disease Control and Prevention, Division of Bacterial Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150069, China.
³ State 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, Northeast Agricultural University, Harbin, 150030, China.
⁴ Section of Paediatric Infectious Disease, Department of Infectious Disease, Imperial College London, St. Mary's Campus, London, W2 1NY, United Kingdom.
⁵ State Key Laboratory for Animal Disease Control and Prevention, Division of Bacterial Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150069, China. Electronic address: Liusiguo@caas.cn.
⁶ State Key Laboratory for Animal Disease Control and Prevention, Division of Bacterial Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150069, China. Electronic address: ligang@caas.cn.

PMID: 37734291
DOI: 10.1016/j.talanta.2023.125202

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.

MeSH terms

Animals
CRISPR-Cas Systems
Humans
Serogroup
Serotyping / methods
Streptococcal Infections* / diagnosis
Streptococcal Infections* / epidemiology
Streptococcal Infections* / veterinary
Streptococcus suis* / genetics
Swine