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使用聚合酶链反应和属特异性寡核苷酸引物检测马粪便中的肠炎沙门氏菌。

Detection of Salmonella enteritidis in equine feces using the polymerase chain reaction and genus-specific oligonucleotide primers.

作者信息

Cohen N D, Wallis D E, Neibergs H L, Hargis B M

机构信息

Department of Large Animal Medicine & Surgery, College of Veterinary Medicine, Texas A&M University, College Station 77843, USA.

出版信息

J Vet Diagn Invest. 1995 Apr;7(2):219-22. doi: 10.1177/104063879500700209.

Abstract

Salmonella was identified in feces from horses, using the polymerase chain reaction (PCR) and genus-specific oligonucleotide primers. Feces from healthy horses were determined to be culture negative and PCR negative for Salmonella. Fecal samples were inoculated with known numbers of colony-forming units (CFU) of S. enteritidis. The fecal samples were enriched overnight in tetrathionate broth, and then DNA was extracted and amplified by PCR using genus-specific primers. Sensitivity of the assay extended to 10 degrees CFU Salmonella enteritidis/g feces; sensitivity of microbiologic culture with enrichment extended to 10 degrees CFU Salmonella enteritidis/g feces. Feces that were not inoculated with S. enteritidis were negative by the PCR. Detection of salmonellae in feces was possible using the PCR within 24 hours from the time of submission of samples. Because samples were enriched, isolates were available for determining antibiograms and serologic grouping or typing.

摘要

使用聚合酶链反应(PCR)和属特异性寡核苷酸引物,在马的粪便中鉴定出了沙门氏菌。经测定,健康马的粪便沙门氏菌培养阴性且PCR阴性。向粪便样本接种已知数量的肠炎沙门氏菌菌落形成单位(CFU)。粪便样本在四硫磺酸盐肉汤中富集过夜,然后提取DNA并使用属特异性引物通过PCR进行扩增。该检测方法的灵敏度可达到每克粪便10个CFU肠炎沙门氏菌;富集后的微生物培养灵敏度可达到每克粪便10个CFU肠炎沙门氏菌。未接种肠炎沙门氏菌的粪便PCR检测呈阴性。从提交样本之时起,在24小时内使用PCR能够检测出粪便中的沙门氏菌。由于样本经过富集,可获得用于确定抗菌谱以及血清学分型或分组的分离株。

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