| 双重PCR检测沙门氏菌和变形杆菌方法的建立 |
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| 引用本文： | 郇 娟郇 娟1,戈红雨2,张成龙1,韩双羽1,师 伟1,王玉宝1. 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, 0(3): 84-86 |
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| 作者姓名： | 郇 娟郇 娟1 戈红雨2 张成龙1 韩双羽1 师 伟1 王玉宝1 |
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| 作者单位： | ( 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) |
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| 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。
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| 关 键 词： | 沙门氏菌 变形杆菌 双重PCR 生化反应 |
Establishment of double PCR method for detection of Salmonella and Proteus |
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| HUAN Juan' target="_blank" rel="external">HUAN Juan1,GE Hong-yu2,ZHANG Cheng-long1,HAN Shuang-yu1,SHI Wei1,WANG Yu-bao2. Establishment of double PCR method for detection of Salmonella and Proteus[J]. Journal of Tianjin Medical University, 2015, 0(3): 84-86 |
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| Authors: | HUAN Juan' target=" _blank" rel=" external" >HUAN Juan1,GE Hong-yu2,ZHANG Cheng-long1,HAN Shuang-yu1,SHI Wei1,WANG Yu-bao2 |
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| Affiliation: | （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） |
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| 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. Results:Thirty-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. Conclusion:The invA-tuf double PCR assay could make rapid and reliable identification of the Salmonella.spp and Proteus.spp isolated on the Salmonella Shigella agar . |
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| Keywords: | Salmonella Proteus double PCR biochemical tests |
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