Merk S, Meyer H, Greiser-Wilke I, Sprague L D, Neubauer H
Institut fuer Medizinische Informatik und Biomathematik, Domagkstrasse 9, 48149 Munster, Germany.
J Vet Med B Infect Dis Vet Public Health. 2006 Aug;53(6):281-5. doi: 10.1111/j.1439-0450.2006.00956.x.
Bacterial DNA (Burkholderia cepacia) was prepared from artificially infected equine ethylenediaminetetraacetic acid (EDTA)-blood and lung tissue by using four standard methods (lysis buffer containing proteinase K, phenol/chloroform/isoamylalcohol-extraction, microwave-treatment, heat treatment) and six commercially available kits (Puregene, High Pure PCR Template Preparation Kit, InstaGene, QiaAmp Tissue Kit, DNAzol and Elu-Quik). After a subsequent polymerase chain reaction (PCR), their efficacy and sensitivity were compared. Concerning the detection limits, the simple lysis with a proteinase K-containing buffer led to the best results for EDTA-blood as well as for artificially infected lung tissue.
通过使用四种标准方法(含蛋白酶K的裂解缓冲液、苯酚/氯仿/异戊醇提取、微波处理、热处理)和六种市售试剂盒(Puregene、高纯PCR模板制备试剂盒、InstaGene、QiaAmp组织试剂盒、DNAzol和Elu-Quik),从人工感染的马乙二胺四乙酸(EDTA)血液和肺组织中制备细菌DNA(洋葱伯克霍尔德菌)。在随后的聚合酶链反应(PCR)之后,比较了它们的功效和灵敏度。关于检测限,用含蛋白酶K的缓冲液进行简单裂解对EDTA血液以及人工感染的肺组织产生了最佳结果。