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A one-step closed-tube enzyme-activated blocked probe assay based on SNP for rapid detection of Salmonella Pullorum

文献类型： 外文期刊

作者： Wen, Junping 1 ; Gou, Hongchao 7 ; Liu, Jing 1 ; Zhou, Hualiang 8 ; Lin, Qijie 1 ; Qu, Xiaoyun 1 ; Chen, Kaifeng 1 ; Wang 1 ;

作者机构： 1.Natl & Reg Joint Engn Lab Medicament Zoonoses Pre, Guangzhou 510642, Peoples R China

2.Minist Agr, Key Lab Zoonoses, Guangzhou 510642, Peoples R China

3.Key Lab Zoonoses Prevent & Control Guangdong Prov, Guangzhou 510642, Peoples R China

4.Minist Agr, Key Lab Anim Vaccine Dev, Guangzhou 510642, Peoples R China

5.Guangdong Lab Lingnan Modern Agr, Guangzhou 510642, Peoples R China

6.South China Agr Univ, Coll Vet Med, Guangzhou 510642, Peoples R China

7.Guangdong Acad Agr Sci, Inst Anim Hlth, Guangzhou 510640, Peoples R China

8.Anim & Plant Inspect & Quarantine Technol Ctr, Shenzhen 518054, Peoples R China

关键词： Salmonella Pullorum; SNP; enzyme-activated blocked probe (EA probe); rapid detection

期刊名称：POULTRY SCIENCE （ 影响因子：3.352； 五年影响因子：3.679 ）

ISSN：

年卷期： 2021 年 100 卷 2 期

页码：

收录情况： SCI

摘要： Salmonella enterica serovar Gallinarum biovars Pullorum (S. Pullorum) is an infectious bacterial pathogen in the poultry industry that causes systemic pullorum disease. This disease causes great losses in terms of the clinical production and quality of chicken products in breeding farms. However, an acknowledged usable rapid detection method for its specific identification has not been reported, and it is generally difficult to distinguish from fowl typhoid caused by Salmonella enterica serovar Gallinarum biovars Gallinarum. The development of a specific and rapid detection method for this pathogen is therefore needed. In the present study, we targeted the single-nucleotide mutation position 237 of the S. Pullorum rfbS gene to develop an enzyme activated blocked probe for its clinical rapid detection. The method displayed robust specificity and reproducibility, and it achieved minimal detection limits of 21 copies/mL of copy number and 4.53 pg/mL of genomic DNA. Compared with traditional identification and PCR methods, this method performed better for the detection of 100 clinical actual samples and without false negative results. The entire process can be accomplished in a 1-step closed-tube operation, overcomes the difficulties currently associated with S. Pullorum detection, and provides a specific and rapid method with broad application potential for SNP detection.

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