|本期目录/Table of Contents|

[1]郇 娟,戈红雨,张成龙,等.双重PCR检测沙门氏菌和变形杆菌方法的建立[J].天津医科大学学报,2015,21(03):84-86.
 HUAN Juan,GE Hong-yu,ZHANG Cheng-long,et al.Establishment of double PCR method for detection of Salmonella and Proteus[J].Journal of Tianjin Medical University,2015,21(03):84-86.
点击复制

双重PCR检测沙门氏菌和变形杆菌方法的建立(PDF)
分享到:

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
21卷
期数:
2015年03期
页码:
84-86
栏目:
技术与方法
出版日期:
2015-05-20

文章信息/Info

Title:
Establishment of double PCR method for detection of Salmonella and Proteus
文章编号:
1006-8147(2015)01-0084-03
作者:
 郇 娟1戈红雨2张成龙1韩双羽1师 伟1王玉宝1
( 1.天津医科大学第二医院感染性疾病研究所,天津 300211;2天津医科大学总医院感染科,天津300052)
Author(s):

HUAN Juan1GE Hong-yu2ZHANG Cheng-long1HAN Shuang-yu1SHI Wei1WANG Yu-bao2

(1 .Institute of Infection Disease,The Second Hospital,Tianjin Medical University, Tianjin 300211,China;2. Department of Infection Disease, General Hospital, Tianjin Medical University ,Tianjin 300052,China)

关键词:
沙门氏菌变形杆菌双重PCR生化反应
Keywords:
SalmonellaProteusdouble PCRbiochemical tests
分类号:
R372
DOI:
-
文献标志码:
A
摘要:
目的:建立快速鉴定腹泻患者粪便标本中和变形杆菌的双重聚合酶链式反应(PCR)方法。方法:针对沙门氏菌属特异基因invA和变形杆菌属特异基因tuf分别设计特异性引物,2013年5月-10月天津某医院肠道门诊腹泻患者粪便标本经SS培养基分离培养后,对可疑菌株同时进行invA-tuf基因双重PCR鉴定和生化鉴定,比较两种方法结果的一致性;对沙门氏菌进一步行血清学分型。结果:双重PCR和生化反应都各自鉴定得到沙门氏菌31株和变形杆菌62株,而且二者鉴定结果完全一致;31株沙门氏菌包括鼠伤寒、肠炎等常见血清型和格兰扁、菲尔摩雷等罕见血清型,均能扩增出283 bp长度片段,62株变形杆菌包括48株奇异变形杆菌、8株普通变形杆菌、2株潘氏变形杆菌、4株产黏液变形杆菌,均能扩增出541bp长度片段。 结论invA-tuf基因双重PCR方法能够快速可靠地鉴定SS培养基上生长的沙门氏菌和变形杆菌。
Abstract:

Objective: To develop a double PCR assay for simultaneous identification of Salmonella and Proteus isolated from stool samples of patients with diarrhea. Methods: Based on the invA gene in Salmonella and the tuf gene in Proteus, 2 pairs of primers were designed. The stool samples of diarrhea patients in the diarrhea outpatient clinic of a hospital in Tianjin from May to October 2013 were collected for the culture on the Salmonella Shigella agar after which double PCR and biochemical identification were carried out simultaneously with the suspected colonies. The consistency of the two methods was evaluated. The serological typing was carried out for Salmonella. ResultsThirty-one strains of Salmonella and 62 strains of Proteus were detected by Double PCR and biochemical identification respectively, and the results were identical for the two methods. The 31 strains of Salmonella included both the common serotypes(e.g. S.Typhimurim and S.Enteritidis)and rare serotypes(e.g. S.Grampian and S.Fillmore). 283bp and 541bp fragments were obtained for all Salmonella and 62 strains of Proteus (48 P.mirabilis,8 P.vulgaris,2 P.penneri and 4 P.myxofaciens) ,respectively. ConclusionThe invA-tuf double PCR assay could make rapid and reliable identification of the Salmonella.spp and Proteus.spp isolated on the Salmonella Shigella agar .

参考文献/References:

[1]Barton Behravesh C, Jones T F, Vugia D J, et al. Deaths associated with bacterial pathogens transmitted commonly through food: foodborne diseases active surveillance network (FoodNet), 1996-2005[J]. J Infect Dis, 2011, 204(2): 263

[2]毛雪丹,胡俊峰,刘秀梅.2003-2007年中国1060起细菌性食源性疾病流行病学特征分析[J].中国食品卫生杂志,2010,22(3):224

[3]王锦彤,钟广辉,熊定凯,等.珠江水中检出与沙门氏菌具有共同抗原的奇异变形杆菌[J].口岸卫生控制,2008,13(2):23

[4]Dione M M, Ikumapayi U, Saha D, et al. Antimicrobial resistance and virulence genes of non-typhoidal Salmonella isolates in The Gambia and Senegal[J]. J Infect Dev Ctries, 2011, 5(11): 765

[5]毕水莲,唐书泽,陈守义,等.食源性变形杆菌属PCR检测方法[J].食品研究与开发,2010,31(2):155

[6]Suez J, Porwollik S, Dagan A, et al. Virulence gene profiling and pathogenicity characterization of non-typhoidal Salmonella accounted for invasive disease in humans[J]. PLoS One, 2013, 8(3): e58449

[7]Rahn K, De Grandis S A, Clarke R C, et al. Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella[J]. Mol Cell Probes, 1992, 6(4): 271

[8]Paradis S, Boissinot M, Paquette N, et al. Phylogeny of the enterobacteriaceae based on genes encoding elongation factor Tu and F-ATPase beta-subunit[J]. Int J Syst Evol Microbiol, 2005, 55(Pt 5): 2013

[9]王慧,朱瑞良,谭燕玲,等.多重PCR检测三种重要食源性致病菌方法的建立及应用[J].中国农业科学,2011,44(11):2334

相似文献/References:

[1]郇 娟,戈红雨,张成龙,等.双重PCR检测沙门氏菌和变形杆菌方法的建立[J].天津医科大学学报,2015,21(01):84.
 HUAN Juan,GE Hong-yu,ZHANG Cheng-long,et al.Establishment of double PCR method for detection of Salmonella and Proteus[J].Journal of Tianjin Medical University,2015,21(03):84.

备注/Memo

备注/Memo:

基金项目  天津市卫生局科研基金项目(2013KZ112)

作者简介 郇娟(1988-),女,硕士在读,研究方向:细菌耐药及致病机制;通信作者:王玉宝,E-mail:wyb2046@163.com

更新日期/Last Update: 2015-01-22