Englen M D, Kelley L C
Poultry Microbiological Safety Research Unit, Richard B. Russell Agricultural Research Center, USDA Agricultural Research Service, Athens, GA 30604-5677, USA.
Lett Appl Microbiol. 2000 Dec;31(6):421-6. doi: 10.1046/j.1365-2672.2000.00841.x.
We have developed an efficient process for rapidly isolating campylobacter DNA using mechanical disruption combined with the guanidine-based reagent DNAzol. Template DNA was isolated by this method from cultures of Campylobacter jejuni resistant to lysis by boiling or enzymes and identified following polymerase chain reaction (PCR) amplification using primers specific for the hippuricase gene. Direct detection of campylobacters in poultry-processing samples by PCR is demonstrated in chicken carcass rinses spiked with lysis-resistant C. jejuni. Our results indicate that this method of DNA isolation may be ideal for direct PCR detection of pathogenic bacteria in complex samples of widely varied origin, especially when the target organisms are difficult to lyse by other means.
我们开发了一种高效的方法,通过机械破碎结合基于胍的试剂DNAzol快速分离弯曲杆菌DNA。通过这种方法从对煮沸或酶解具有抗性的空肠弯曲菌培养物中分离模板DNA,并使用针对马尿酸酶基因的引物进行聚合酶链反应(PCR)扩增后进行鉴定。在接种了抗裂解空肠弯曲菌的鸡胴体冲洗液中,通过PCR直接检测家禽加工样品中的弯曲杆菌。我们的结果表明,这种DNA分离方法可能是直接PCR检测来自广泛不同来源的复杂样品中病原菌的理想方法,特别是当目标生物体难以通过其他方法裂解时。