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沙门氏菌LAMP可视化检测方法的建立 预览 被引量:1

Development of a Visual LAMP Test Method for Samonella
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摘要 为实现沙门氏菌的快速现场检测,根据沙门氏菌invA基因编码区序列,设计合成1套引物,通过对反应物浓度和反应条件优化,建立了沙门氏菌属特异性LAMP检测方法;通过对invA基因8个区域的6条引物配对进行等温扩增,并在反应体系中加入HNB来指示反应结果,实现了反应的快速闭管检测。灵敏度试验显示,建立的方法可以检出稀释至3.6×101CFU/mL菌液所提取的DNA;特异性试验显示,建立的方法与大肠杆菌DNA、志贺氏菌DNA、布鲁氏菌试管凝集抗原DNA、炭疽沉淀抗原DNA以及猪链球菌2型DNA不发生交叉反应,但能检出鸡白痢、禽伤寒沙门氏菌凝集抗原以及马流产试管凝集抗原提取的DNA;重复性试验显示,3个稀释度菌液DNA的3次重复检测均为阳性;应用试验显示,从经增菌培养的200份进境猴粪拭子样品中检出阳性2份,与细菌分离鉴定检测结果一致。结果表明,所建立的方法敏感性和特异性较高,重复性满足实际要求,可用于进境动物样品的沙门氏菌检测。 In searching of rapid field detection methods for salmonella,a set of primers were designed and synthesized–which were 3 pairs of primers that identifying 8 distinct regions of invA gene of Salmonella for isothermal amplification.With optimizing the reactant concentration and reaction conditions,and by adding HNB to the reaction as an indicator,a closed-tube LAMP method for rapid field detection of Salmonella was developed.Sensitivity test showed that the DNAs extracted from 3.6×101 CFU/mL bacterial suspension could be detected.Specificity test confirmed that no cross reaction was found between the method with the DNAs of Golibacillus,Shigella,Brucella agglutinating antigen,anthrax precipitation antigen and type 2 Streptococcus suis,but it could detect DNAs from pullorosis,Salmonella typhi agglutinating antigen and Abortus-equi agglutination antigen.Repetitive test verified that the results of 3 experiments for DNAs in 3 diluted bacteria solutions were all positive.Application test indicated that 2 of 200 swab samples from manures of imported monkeys were found positive,which was identical with the result of bacterial isolation test.All test results proved that the method is featured with higher degree of sensitivity and specificity,and its repeatability allows practical application,hence it would be a better approach to detect Salmonella in animal import quarantine.
作者 高志强 汪琳 张灿 任彤 赵相鹏 尹羿 蒲静 史雅然 刘凤华 Gao Zhiqiang;Wang Lin;Zhang Can;Ren Tong;Zhao Xiangpeng;Yin Yi;Pu Jing;Shi Yaran;Liu Fenghua(Beijing Inspection and Quarantine Testing Center,Beijing 100026,China;Institute for Disease Prevention and Control of Chinese PLA,Beijing 100071,China;Beijing College of Agriculture,Beijing 102206,China)
出处 《中国动物检疫》 CAS 2019年第2期68-72,共5页 China Animal Health Inspection
基金 国家重点研发计划项目(2017YFF0210305) 北京市科技计划课题(Z171100001317011).
关键词 沙门氏菌 LAMP 可视化检测 检疫 Salmonella LAMP visual detection quarantine
作者简介 通信作者:张灿.
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  • 1陈金顶,索青利,廖明,辛朝安.沙门氏菌的invA基因序列分析与分子检测[J].中国人兽共患病杂志,2004,20(10):868-871. 被引量:46
  • 2Anonymous. The community summary report on trends and sources of zoonoses , zoonotic agents , antimicrobial resistance and foodbome outbreaks in the European Union in 2006 [J]. The EFSA Journal, 2006, 130: 1-352. 被引量:1
  • 3Byme D. Zoonoses: Commissioner David Byrne wel- comes new legislation to combat food-borne diseases such as Salmonella (IP/03/1306) [R]. European Commis- sion, 2003. 被引量:1
  • 4Nowak B, von Mtiffiing T, Chaunchom S, et al. Sal- monella contamination in pigs at slaughter and on the farm: A field study using an antibody ELISA test and a PCR technique [J]. International Journal of Food Mi- crobiology. 2007, 115(3) : 259-267. 被引量:1
  • 5McGuinness S, McCabe Evonne, O'Regan Edel, et al.Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella onfresh meat[J]. Meat Science, 2009, 83 (3) : 555-562. 被引量:1
  • 6Ihira Masaru, Ohta Akane, Sugata Ken, et al. Loop- mediated isothermal amplification for discriminating be- tween human herpesvirus 6 A and B [J]. Journal of Vi- rologicalMethods, 2008, 154(1/2): 223-225. 被引量:1
  • 7Curtis K A, Rudolph D L, Owen S M. Rapid detection of HIV-1 by reverse transcription, loop-mediated iso- thermal amplification(RT-LAMP)[J]. Journal of Vi- rologicalMethods, 2008, 151(2) : 264-270. 被引量:1
  • 8Gao Hongwei, Lei Zhiwen, Jia Juntao, et al. Applica- tion of loop-mediated isothermal amplification for detec- tion of Yersinia enterocolitica in pork meat [J]. Journal of Microbiological Methods, 2008, 77 (2) : 198-201. 被引量:1
  • 9Mori Y, Nagamine K, Tomita N, et al. Detection of loop-mediate isothermal amplification reaction by turbid- ity derived from magnesium pyrophosphate formation[J]. Biochem Biophys Res Commun, 2001, 289(1) : 150-154. 被引量:1
  • 10Btilte M, Jakob E The use of a PCR-generated invA probe for the detection of Salmonella spp. in artificially and naturally contaminated foods [J]. International Journal of Food Microbiology, 1995, 26(3) : 335- 344. 被引量:1

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