摘要
目的建立一种检测奇异变形杆菌(Proteus mirabilis)快速且特异的PCR方法。方法针对编码奇异变形杆菌脲酶调节子(ureR)的基因序列设计2条PCR引物,扩增片段长度为225bp;通过对2株奇异变形杆菌和14株非奇异变形杆菌进行PCR检测、利用该PCR方法和传统分离培养法同时对60份食物中毒送检标本进行检测来评价其特异性;将奇异变形杆菌菌液做一系列10倍稀释后进行PCR检测来对其进行敏感性分析。结果2株奇异变形杆菌出现PCR扩增反应,扩增产物经测序鉴定正确,14株非奇异变形杆菌没有出现PCR扩增反应,且PCR检测奇异变形杆菌的结果与传统培养方法一致,PCR方法相比传统培养方法更快速,在4h内即可完成扩增反应;PCR方法敏感性为30cfu/ml。结论建立了一种快速、准确检测奇异变形杆菌的PCR方法,适合奇异变形杆菌日常监测和快速诊断的需要。
Objective To develop a POR assay for the rapid and specific detection of Proteus mirabilis. Methods According to the sequences of ureR gene of Proteus mirabilis, 2 primers were designed and used for PCR. The length of amplicon was 225 bp. To evaluate the specificity of POR assay, two strains of Proteus mirabilis and 14 strains of non - Proteus mirabilis were tested by PCR, then 60 specimens of foodborne disease were tested by POR and Plate - culture method simultaneously. One strain of Proteus mirabilis was 10 -fold serially diluted and was amplified by PCR to verify detection limit. Results With 2 strains of Proteus mirabilis, the results of PCR assay were observed and the POR products were also confirmed by DNA sequence analysis. Amplification was not observed when 14 strains of non Proteus mirabilis were tested. The results of the PCR and Plate --- culture method were the same, so the specificity of PCR was confirmed. In addition, PCR detected Proteus mirabilis within 4 h and thus POR is superior to Plate culture method in terms of rapidity, and the detection limit of the PCR assay was 30 cfu/ml of Proteus mirabilis. Conclusions These results indicate that POR assay is a rapid, specific and sensitive detection method for Proteus mirabilis, and it is suitable for daily monitoring and rapid diagnosis of Proteus mirabilis.
出处
《实用预防医学》
CAS
2009年第2期560-562,共3页
Practical Preventive Medicine
作者简介
林红乐，女，主管检验师，主要从事微生物检验工作。
