Frontiers in Cellular and Infection Microbiology (Nov 2022)
Accelerated loop-mediated isothermal amplification method for the rapid detection of Streptococcus suis serotypes 2 and 14 based on single nucleotide polymorphisms
- Jiajia Meng,
- Chunling Li,
- Chunling Li,
- Chunling Li,
- Chunling Li,
- Yu Wang,
- Zhibiao Bian,
- Zhibiao Bian,
- Zhibiao Bian,
- Zhibiao Bian,
- Pinpin Chu,
- Pinpin Chu,
- Pinpin Chu,
- Pinpin Chu,
- Shaolun Zhai,
- Shaolun Zhai,
- Shaolun Zhai,
- Shaolun Zhai,
- Dongxia Yang,
- Dongxia Yang,
- Dongxia Yang,
- Dongxia Yang,
- Shuai Song,
- Shuai Song,
- Shuai Song,
- Shuai Song,
- Yan Li,
- Yan Li,
- Yan Li,
- Yan Li,
- Zhiyong Jiang,
- Zhiyong Jiang,
- Zhiyong Jiang,
- Zhiyong Jiang,
- Kunli Zhang,
- Kunli Zhang,
- Kunli Zhang,
- Kunli Zhang,
- Yugu Li,
- Hongchao Gou,
- Hongchao Gou,
- Hongchao Gou,
- Hongchao Gou
Affiliations
- Jiajia Meng
- Guangdong Laboratory for Lingnan Modern Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
- Chunling Li
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Chunling Li
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Chunling Li
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Chunling Li
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Yu Wang
- Guangdong Laboratory for Lingnan Modern Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
- Zhibiao Bian
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Zhibiao Bian
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Zhibiao Bian
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Zhibiao Bian
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Pinpin Chu
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Pinpin Chu
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Pinpin Chu
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Pinpin Chu
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Shaolun Zhai
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Shaolun Zhai
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Shaolun Zhai
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Shaolun Zhai
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Dongxia Yang
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Dongxia Yang
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Dongxia Yang
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Dongxia Yang
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Shuai Song
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Shuai Song
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Shuai Song
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Shuai Song
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Yan Li
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Yan Li
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Yan Li
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Yan Li
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Zhiyong Jiang
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Zhiyong Jiang
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Zhiyong Jiang
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Zhiyong Jiang
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Kunli Zhang
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Kunli Zhang
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Kunli Zhang
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Kunli Zhang
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- Yugu Li
- Guangdong Laboratory for Lingnan Modern Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China
- Hongchao Gou
- Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China
- Hongchao Gou
- Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China
- Hongchao Gou
- Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming, China
- Hongchao Gou
- Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou, China
- DOI
- https://doi.org/10.3389/fcimb.2022.1034762
- Journal volume & issue
-
Vol. 12
Abstract
Streptococcus suis serotypes 2 and 14 are the most prevalent zoonotic strains. The establishment of a sensitive and extremely accurate method for point-of-care testing for Streptococcus suis serotype 2 and 14 strains is highly desirable. In this study, a loop primer probe-introduced loop-mediated isothermal amplification assay was developed to differentiate Streptococcus suis serotypes 2 and 14 based on SNP (single nucleotide polymorphism). The specific fluorescent probes were designed for the SNP site specific for serotype 2 and 14 Streptococcus suis cpsK genes, and the loop primer probe-introduced loop-mediated isothermal amplification (LAMP) assay was developed using the specific cleavage properties of the RNase H2 enzyme. Rapid and efficient LAMP assays were realized through the use of loop forward primers and stem forward primers. The results showed that the amplification reaction can be performed efficiently at 59°C. The results can be real-time detected or judged using a smartphone and a 3D-printed visualization cassette. The sensitivity of the LAMP assay can reach 18.4 CFU within 40 minutes. The detection rate of the assay system was evaluated using 19 clinical samples with suspected Streptococcus suis infection, and the detection rate was consistent with the sequencing method, suggesting that the test is highly practical. The LAMP assay for Streptococcus suis serotypes 2 and 14 established in this study has strong specificity, high sensitivity, and simple operation, while the reaction can be performed at an isothermal temperature and is not dependent on complex instruments or professional operators, making it suitable for field testing.
Keywords
- Streptococcus suis serotypes 2 and 14
- single-nucleotide polymorphism
- rapid detection
- RNase H2 enzyme
- loop-mediated isothermal amplification
