Rapid detection of Salmonella enterica serovar Typhimurium DT104 strains by the polymerase chain reaction.
Rapid detection of Salmonella enterica serovar Typhimurium DT104 strains by the polymerase chain reaction.
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DOI：
10.1186/s13028-015-0143-x
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发表时间：
2015-09-25
影响因子：
2.1
通讯作者：
Uchida I
中科院分区：
文献类型：
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作者：
Yukawa S;Tamura Y;Tanaka K;Uchida I
肠道沙门氏菌感染是发达国家的一个主要公共卫生问题，多重耐药菌株已变得越来越普遍。S.肠道血清型鼠伤寒沙门氏菌DT 104（DT 104）菌株在日本的家畜中普遍存在，并且包括多种多重耐药的沙门氏菌菌株。肠对这些菌株的流行病学分析对农业和公共卫生都至关重要;然而，对这些菌株的诊断测试产生了不一致的结果。我们开发了一种快速，简单，廉价的聚合酶链反应检测多药耐药DT 104菌株。我们设计了针对DT 104菌株编码的原噬菌体ST 104序列的特异性引物，并通过测定一组50株肠道沙门氏菌分离株来评估这些引物的特异性。从每个DT 104菌株的基因组中产生预期大小的扩增产物;然而，ST 104引物未能扩增来自非DT 104菌株的肠道沙门氏菌鼠伤寒血清型或其他沙门氏菌的产物。肠道血清型。此外，使用ST 104引物产生的探针通过Southern杂交检测编码DT 104的ST 104区的限制性片段。ST 104引物对DT 104菌株具有特异性，适用于流行病学应用。
Infection with Salmonella enterica is a major public health concern in developed countries, and multidrug-resistant strains have become increasingly prevalent. S. enterica serovar Typhimurium DT104 (DT104) strains are prevalent in livestock in Japan and include numerous strains of multidrug-resistant S. enterica. Epidemiological analysis of these strains is critical for both agriculture and public health; however, diagnostic tests for these strains have yielded inconsistent results. We developed a rapid, simple, and inexpensive polymerase chain reaction test to detect multi-drug resistant DT104 strains. We designed primers specific to the prophage ST104 sequence encoded by DT104 strains and assessed the specificity of these primers by assaying a panel of 50 S.enterica isolates. Amplification products of the expected size were generated from the genomes of each of the DT104 strains; however, the ST104 primers failed to amplify products from non-DT104 strains of S.enterica serovar Typhimurium or other S. enterica serovars. Furthermore, a probe generated using the ST104 primers detected a restriction fragment encoding the ST104 region of DT104 by Southern hybridization. The ST104 primers exhibit specificity to DT104 strains and are suitable for epidemiological applications.