Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method
Abstract Streptococcus suis serotypes 2 and 14 are the most common zoonotic strains, but previous identification methods made distinguish these two serotypes from other S. suis serotypes difficult. To effectively prevent and control them, there is an urgent need for a highly sensitive and specific m...
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2024-11-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-024-04361-9 |
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| author | Yaxing Su Jiajia Meng Mingwei Zhao Chunling Li Shaolun Zhai Yan Li Pinpin Chu Zhibiao Bian Kunli Zhang Dongxia Yang Zhiyong Jiang Hongchao Gou Chenggang Xu |
| author_facet | Yaxing Su Jiajia Meng Mingwei Zhao Chunling Li Shaolun Zhai Yan Li Pinpin Chu Zhibiao Bian Kunli Zhang Dongxia Yang Zhiyong Jiang Hongchao Gou Chenggang Xu |
| author_sort | Yaxing Su |
| collection | DOAJ |
| description | Abstract Streptococcus suis serotypes 2 and 14 are the most common zoonotic strains, but previous identification methods made distinguish these two serotypes from other S. suis serotypes difficult. To effectively prevent and control them, there is an urgent need for a highly sensitive and specific method to identify these two serotypes. In this study, a fluorescent probe was designed for the single nucleotide polymorphism site at cpsK 483 of Streptococcus suis type 2 and type 14 compared with other serotypes, and an enzyme-activated probe quantitative PCR (EA-probe qPCR) method was established for the detection of Streptococcus suis type 2 and type 14 by combining with the specific hydrolysis characteristics of the RNase H2 enzyme. The results showed that the optimal probe concentration for this method was 0.5 µM and the optimal RNase H2 enzyme concentration was 25 mU.This method showed no reactivity with genomic DNA from Streptococcus suis strains 1/2, 5, 7, 9, 23, 28, 29, and 31, confirming its high specificity. And its sensitivity can reach 18.4 CFU. In addition, 19 clinical strains of Streptococcus suis type 2 or type 1/2 were tested. The results showed 100% agreement with the gene sequencing method. In conclusion, this method can meet the needs of accurate laboratory testing of Streptococcus suis serotypes 2 and 14 and has value for clinical prevention. |
| format | Article |
| id | doaj-art-7f50f73e8d0e4f6980f1975d963f29d6 |
| institution | Kabale University |
| issn | 1746-6148 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj-art-7f50f73e8d0e4f6980f1975d963f29d62024-11-10T12:30:00ZengBMCBMC Veterinary Research1746-61482024-11-012011710.1186/s12917-024-04361-9Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR methodYaxing Su0Jiajia Meng1Mingwei Zhao2Chunling Li3Shaolun Zhai4Yan Li5Pinpin Chu6Zhibiao Bian7Kunli Zhang8Dongxia Yang9Zhiyong Jiang10Hongchao Gou11Chenggang Xu12College of Veterinary Medicine, South China Agricultural University, Guangdong Laboratory for Lingnan Modern AgricultureCollege of Veterinary Medicine, South China Agricultural University, Guangdong Laboratory for Lingnan Modern AgricultureCollege of Veterinary Medicine, South China Agricultural University, Guangdong Laboratory for Lingnan Modern AgricultureInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceInstitute of Animal Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangdong Academy of Agricultural Sciences, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong ProvinceCollege of Veterinary Medicine, South China Agricultural University, Guangdong Laboratory for Lingnan Modern AgricultureAbstract Streptococcus suis serotypes 2 and 14 are the most common zoonotic strains, but previous identification methods made distinguish these two serotypes from other S. suis serotypes difficult. To effectively prevent and control them, there is an urgent need for a highly sensitive and specific method to identify these two serotypes. In this study, a fluorescent probe was designed for the single nucleotide polymorphism site at cpsK 483 of Streptococcus suis type 2 and type 14 compared with other serotypes, and an enzyme-activated probe quantitative PCR (EA-probe qPCR) method was established for the detection of Streptococcus suis type 2 and type 14 by combining with the specific hydrolysis characteristics of the RNase H2 enzyme. The results showed that the optimal probe concentration for this method was 0.5 µM and the optimal RNase H2 enzyme concentration was 25 mU.This method showed no reactivity with genomic DNA from Streptococcus suis strains 1/2, 5, 7, 9, 23, 28, 29, and 31, confirming its high specificity. And its sensitivity can reach 18.4 CFU. In addition, 19 clinical strains of Streptococcus suis type 2 or type 1/2 were tested. The results showed 100% agreement with the gene sequencing method. In conclusion, this method can meet the needs of accurate laboratory testing of Streptococcus suis serotypes 2 and 14 and has value for clinical prevention.https://doi.org/10.1186/s12917-024-04361-9Streptococcus suis serotypes 2 and 14Single-nucleotide polymorphismEA-probe qPCRRapid detectionRNase H2 enzyme |
| spellingShingle | Yaxing Su Jiajia Meng Mingwei Zhao Chunling Li Shaolun Zhai Yan Li Pinpin Chu Zhibiao Bian Kunli Zhang Dongxia Yang Zhiyong Jiang Hongchao Gou Chenggang Xu Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method BMC Veterinary Research Streptococcus suis serotypes 2 and 14 Single-nucleotide polymorphism EA-probe qPCR Rapid detection RNase H2 enzyme |
| title | Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method |
| title_full | Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method |
| title_fullStr | Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method |
| title_full_unstemmed | Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method |
| title_short | Rapid detection of zoonotic Streptococcus suis serotype 2 and 14 by enzyme-activated probe fluorescence quantitative PCR method |
| title_sort | rapid detection of zoonotic streptococcus suis serotype 2 and 14 by enzyme activated probe fluorescence quantitative pcr method |
| topic | Streptococcus suis serotypes 2 and 14 Single-nucleotide polymorphism EA-probe qPCR Rapid detection RNase H2 enzyme |
| url | https://doi.org/10.1186/s12917-024-04361-9 |
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