Chen Hongyuan, Zhong Qingping, Wang Li, Sun Yuanming
The Higher Education Key Lab of Guangdong Province Food Quality and Safety, Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
Wei Sheng Yan Jiu. 2010 Sep;39(5):597-600.
Based on the invasive plasmid antigen H gene (ipaH) of S. dysenteriae, one pair of specific primers was designed for PCR assays in this study. The concentrations of dNTP, Mg2+ and primer, dosage of Taq DNA polymerase, annealing temperature and circulating parameter in the PCR amplification system were optimized. In this way, a rapid and stable method of PCR assay for the detection of S. dysenteriae was established. The specificity and sensitivity of PCR were also analyzed. The detection limits of pure culture and genomic DNA in the PCR assay were 1.06 x 10(2) cfu/ml and 106.34 pg/PCR system, respectively. The detection limit for S. dysenteriae in artificially contaminated food samples was 3.21 x 10(4) cfu/ml. These results indicated that the PCR method for S. dysenteriae detection was simple, rapid, high in specificity and sensitivity and suitable for the detection of pathogens in foods caused by Shigella dysenteriae.
基于痢疾志贺氏菌的侵袭性质粒抗原H基因（ipaH），本研究设计了一对特异性引物用于PCR检测。对PCR扩增体系中的dNTP、Mg2+和引物浓度、Taq DNA聚合酶用量、退火温度及循环参数进行了优化。通过这种方式，建立了一种快速、稳定的检测痢疾志贺氏菌的PCR方法。还分析了PCR的特异性和灵敏度。PCR检测中纯培养物和基因组DNA的检测限分别为1.06×10(2) cfu/ml和106.34 pg/PCR体系。人工污染食品样本中痢疾志贺氏菌的检测限为3.21×10(4) cfu/ml。这些结果表明，用于检测痢疾志贺氏菌的PCR方法简单、快速、特异性和灵敏度高，适用于检测由痢疾志贺氏菌引起的食品中的病原体。
