National and Regional Joint Engineering Laboratory For Medicament of Zoonoses Prevention and Control, Guangzhou 510642, China; Key Laboratory of Zoonoses, Ministry of Agriculture, Guangzhou 510642, China; Key Laboratory of Zoonoses Prevention and Control of Guangdong Province, Guangzhou 510642, China; Key Laboratory of Animal Vaccine Development, Ministry of Agriculture, Guangzhou 510642, China; Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, China; College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.
Poult Sci. 2021 Feb;100(2):1059-1067. doi: 10.1016/j.psj.2020.11.007. Epub 2020 Nov 18.
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/μL of copy number and 4.53 pg/μL 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.
鸡白痢沙门氏菌血清型鸡白痢亚种（S. Pullorum）是家禽业中一种传染性细菌性病原体，可引起全身性鸡白痢病。这种疾病在养殖场的临床生产和鸡肉产品质量方面造成了巨大损失。然而，目前尚未报道针对其特异性鉴定的可用快速检测方法，并且通常难以将其与由鸡白痢沙门氏菌血清型鸡白痢亚种引起的禽伤寒区分开来。因此，需要开发针对这种病原体的特异性和快速检测方法。在本研究中，我们针对 S. Pullorum rfbS 基因的单核苷酸突变位置 237 开发了一种酶激活的阻断探针，用于其临床快速检测。该方法显示出强大的特异性和重现性，其检测限最小可达 21 拷贝/μL 的拷贝数和 4.53 pg/μL 的基因组 DNA。与传统的鉴定和 PCR 方法相比，该方法在检测 100 份临床实际样本时表现更好，且没有假阴性结果。整个过程可以在 1 步封闭管操作中完成，克服了当前 S. Pullorum 检测所面临的困难，并为 SNP 检测提供了一种具有广泛应用潜力的特异性和快速方法。
