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口服攻毒后肠炎沙门氏菌在雏鸭内脏器官中的复制动力学:一项使用实时PCR的定量时间进程研究

Replication kinetics of Salmonella enteritidis in internal organs of ducklings after oral challenge: a quantitative time-course study using real-time PCR.

作者信息

Deng S X, Cheng A C, Wang M S, Li X R, Yan B

机构信息

Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Yaan, 625014, Sichuan Province, China.

出版信息

Vet Res Commun. 2009 Mar;33(3):273-80. doi: 10.1007/s11259-008-9175-2. Epub 2008 Sep 9.

Abstract

This research was undertaken to understand the replication kinetics of Salmonella enteritidis (S. enteritidis) in the internal organs of ducklings after oral challenge over a 2 wk period. A serovar-specific real-time, fluorescence-based quantitative polymerase chain reaction (FQ-PCR) assay was used to detect genomic DNA of S. enteritidis in the blood and the internal organs at different time points respectively. The results showed that the spleen was positive at 12 h post inoculation (PI) and the blood was at 14 h PI. The organism was detected in the liver and heart at 16 h PI, the pancreas and kidney were positive at 20 h PI, and the final organ to show a positive results was the gallbladder at 22 h PI. The copy number of S. enteritidis DNA in each tissue reached a peak at 24 h-36 h PI, with the liver and spleen containing the highest concentration of S. enteritidis. The blood, heart, kidney, pancreas, and gallbladder had low concentrations. S. enteritidis populations began to decrease and were not detectable at 3 d PI, but were still present up to 2 wk for the spleen without causing apparent symptoms. To make the results meaningful, a side-by-side bacteriology method (IFA) was performed. The results of IFA were similar to the FQ-PCR assay. This research provided a significant data for understanding the life cycle of S. enteritidis in the internal organs, and may help to understand the pathogenesis of S.entertidis in the future.

摘要

本研究旨在了解口服攻击后2周内肠炎沙门氏菌(肠炎沙门菌)在雏鸭内脏器官中的复制动力学。采用血清型特异性实时荧光定量聚合酶链反应(FQ-PCR)检测不同时间点雏鸭血液和内脏器官中肠炎沙门菌的基因组DNA。结果显示,接种后12小时脾脏呈阳性,14小时血液呈阳性。接种后16小时在肝脏和心脏中检测到该菌,20小时胰腺和肾脏呈阳性,最后显示阳性结果的器官是胆囊,时间为接种后22小时。各组织中肠炎沙门菌DNA拷贝数在接种后24小时至36小时达到峰值,肝脏和脾脏中肠炎沙门菌浓度最高。血液、心脏、肾脏、胰腺和胆囊中的浓度较低。肠炎沙门菌数量在接种后3天开始减少,接种后3天无法检测到,但脾脏中在2周内仍有存在,且未引起明显症状。为使结果有意义,采用了并行细菌学方法(IFA)。IFA结果与FQ-PCR检测结果相似。本研究为了解肠炎沙门菌在内脏器官中的生命周期提供了重要数据,并可能有助于未来了解肠炎沙门菌的发病机制。

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