实时 PCR 检测阪崎肠杆菌。
Detection of Enterobacter sakazakii strains by real-time PCR.
机构信息
Federal Institute for Risk Assessment, Diedersdorfer Weg 1, D-12277 Berlin, Germany.
出版信息
J Food Prot. 2005 Aug;68(8):1623-7. doi: 10.4315/0362-028x-68.8.1623.
A precise 5' nuclease (TaqMan) real-time PCR was developed and validated in house for the specific detection of Enterobacter sakazakii isolates. Specifically designed nonpatented primers and a hydrolysis (TaqMan) probe were used to target the 16S rRNA gene. All 27 E. sakazakii and 141 non-E. sakazakii strains tested with the real-time PCR were identified correctly. To monitor false-negative results, an internal amplification control was coamplified with the same primers used for the E. sakazakii DNA. The detection probability of the assay was 56% when an E. sakazakii cell suspension containing 10(2) CFU/ml was used as template in the PCR (0.5 CFU per reaction) and 100% with a 10(3) CFU/ml suspension. This PCR assay should be very useful for the diagnostic detection of E. sakazakii in foods, especially powdered infant formula, after cultural enrichment.
建立并验证了一种针对阪崎肠杆菌分离株的特异性检测的精确 5' 核酸酶（TaqMan）实时 PCR 方法。该方法使用特异性设计的非专利引物和水解（TaqMan）探针靶向 16S rRNA 基因。用实时 PCR 对 27 株阪崎肠杆菌和 141 株非阪崎肠杆菌进行检测，结果均正确识别。为了监测假阴性结果，与用于阪崎肠杆菌 DNA 的相同引物一起共扩增了内部扩增对照。当将含有 10(2)CFU/ml 的阪崎肠杆菌细胞悬浮液用作 PCR 模板（每个反应 0.5 CFU）时，该检测方法的检测概率为 56%，当使用 10(3)CFU/ml 悬浮液时，检测概率为 100%。该 PCR 检测方法对于食品（尤其是粉状婴儿配方奶粉）中经文化富集后的阪崎肠杆菌的诊断检测非常有用。
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