An SNP-based diagnostic method for Brucella S2 vaccine strain infections
BackgroundBrucellosis, a zoonotic bacterial infection caused by Brucella species, exhibits a global distribution. The Brucella S2 vaccine strain is known to cause brucellosis. Current serological antibody assays cannot distinguish between infections caused by the S2 strain and those caused by wild-t...
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| Language: | English |
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Frontiers Media S.A.
2025-06-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2025.1570220/full |
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| author | Xingya Wang Xingya Wang Xiaowei Tian Xiaowei Tian Wanyang Li Yuanchao Yang Shuai Zhang Hui Wang Wanru Geng Wanru Geng Jingbo Zhai Jingbo Zhai Jingbo Zhai |
| author_facet | Xingya Wang Xingya Wang Xiaowei Tian Xiaowei Tian Wanyang Li Yuanchao Yang Shuai Zhang Hui Wang Wanru Geng Wanru Geng Jingbo Zhai Jingbo Zhai Jingbo Zhai |
| author_sort | Xingya Wang |
| collection | DOAJ |
| description | BackgroundBrucellosis, a zoonotic bacterial infection caused by Brucella species, exhibits a global distribution. The Brucella S2 vaccine strain is known to cause brucellosis. Current serological antibody assays cannot distinguish between infections caused by the S2 strain and those caused by wild-type Brucella.ObjectiveTo develop a diagnostic method capable of specifically detecting S2 vaccine strain infections.MethodsTwo probes were designed targeting single nucleotide polymorphism (SNP) loci upstream of the sugar ABC gene; quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) methods were established. The performances of these methods were evaluated. The transient stem-loop structure of the DNA template was predicted, and the impact of probe overlap with the transient stem-loop structure on detection sensitivity was analyzed. Clinical applicability was assessed using 50 blood samples from brucellosis patients.ResultsBoth types of methods demonstrated high specificity. However, MGB-SNPdd showed greater sensitivity than other detection methods. Reduction of overlap between the probe sequence and the transient stem-loop structure enhanced detection sensitivity. In the clinical applicability analysis, ddPCR methods exhibited higher rates of S2 vaccine strain detection compared with qPCR methods.ConclusionSNP-based ddPCR methods demonstrate higher sensitivity than qPCR methods and enable specific detection of brucellosis caused by the S2 vaccine strain. Reduction of probe overlap with the transient stem-loop structure improves detection sensitivity, providing valuable insights for enhanced PCR amplification efficiency. |
| format | Article |
| id | doaj-art-8db2ad1a65244e4f8f33540e8f1e91a6 |
| institution | Kabale University |
| issn | 2297-1769 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj-art-8db2ad1a65244e4f8f33540e8f1e91a62025-08-20T03:30:44ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-06-011210.3389/fvets.2025.15702201570220An SNP-based diagnostic method for Brucella S2 vaccine strain infectionsXingya Wang0Xingya Wang1Xiaowei Tian2Xiaowei Tian3Wanyang Li4Yuanchao Yang5Shuai Zhang6Hui Wang7Wanru Geng8Wanru Geng9Jingbo Zhai10Jingbo Zhai11Jingbo Zhai12School of Basic Medical Sciences, Inner Mongolia Minzu University, Tongliao, ChinaDepartment of Laboratory, Hulunbuir Second People’s Hospital, Hulunbuir, ChinaDepartment of Intensive Medicine, Affiliated Hospital of Inner Mongolia Minzu University, Tongliao, ChinaCollege of Clinical Medicine, Inner Mongolia Minzu University, Tongliao, ChinaSchool of Basic Medical Sciences, Inner Mongolia Minzu University, Tongliao, ChinaSchool of Basic Medical Sciences, Inner Mongolia Minzu University, Tongliao, ChinaSchool of Basic Medical Sciences, Inner Mongolia Minzu University, Tongliao, ChinaSchool of Basic Medical Sciences, Inner Mongolia Minzu University, Tongliao, ChinaDepartment of Intensive Medicine, Affiliated Hospital of Inner Mongolia Minzu University, Tongliao, ChinaCollege of Clinical Medicine, Inner Mongolia Minzu University, Tongliao, ChinaSchool of Basic Medical Sciences, Inner Mongolia Minzu University, Tongliao, ChinaKey Laboratory of Zoonose Prevention and Control at Universities of Inner Mongolia Autonomous Region, Tongliao, ChinaBrucellosis Prevention and Treatment Engineering Research Center of Inner Mongolia Autonomous Region, Tongliao, ChinaBackgroundBrucellosis, a zoonotic bacterial infection caused by Brucella species, exhibits a global distribution. The Brucella S2 vaccine strain is known to cause brucellosis. Current serological antibody assays cannot distinguish between infections caused by the S2 strain and those caused by wild-type Brucella.ObjectiveTo develop a diagnostic method capable of specifically detecting S2 vaccine strain infections.MethodsTwo probes were designed targeting single nucleotide polymorphism (SNP) loci upstream of the sugar ABC gene; quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) methods were established. The performances of these methods were evaluated. The transient stem-loop structure of the DNA template was predicted, and the impact of probe overlap with the transient stem-loop structure on detection sensitivity was analyzed. Clinical applicability was assessed using 50 blood samples from brucellosis patients.ResultsBoth types of methods demonstrated high specificity. However, MGB-SNPdd showed greater sensitivity than other detection methods. Reduction of overlap between the probe sequence and the transient stem-loop structure enhanced detection sensitivity. In the clinical applicability analysis, ddPCR methods exhibited higher rates of S2 vaccine strain detection compared with qPCR methods.ConclusionSNP-based ddPCR methods demonstrate higher sensitivity than qPCR methods and enable specific detection of brucellosis caused by the S2 vaccine strain. Reduction of probe overlap with the transient stem-loop structure improves detection sensitivity, providing valuable insights for enhanced PCR amplification efficiency.https://www.frontiersin.org/articles/10.3389/fvets.2025.1570220/fullbrucellosis diagnosisBrucella suis S2qPCRdigital PCRnucleic acid secondary structureSNP locus |
| spellingShingle | Xingya Wang Xingya Wang Xiaowei Tian Xiaowei Tian Wanyang Li Yuanchao Yang Shuai Zhang Hui Wang Wanru Geng Wanru Geng Jingbo Zhai Jingbo Zhai Jingbo Zhai An SNP-based diagnostic method for Brucella S2 vaccine strain infections Frontiers in Veterinary Science brucellosis diagnosis Brucella suis S2 qPCR digital PCR nucleic acid secondary structure SNP locus |
| title | An SNP-based diagnostic method for Brucella S2 vaccine strain infections |
| title_full | An SNP-based diagnostic method for Brucella S2 vaccine strain infections |
| title_fullStr | An SNP-based diagnostic method for Brucella S2 vaccine strain infections |
| title_full_unstemmed | An SNP-based diagnostic method for Brucella S2 vaccine strain infections |
| title_short | An SNP-based diagnostic method for Brucella S2 vaccine strain infections |
| title_sort | snp based diagnostic method for brucella s2 vaccine strain infections |
| topic | brucellosis diagnosis Brucella suis S2 qPCR digital PCR nucleic acid secondary structure SNP locus |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2025.1570220/full |
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