Li Y, Mustapha A
Department of Food Science, University of Missouri, Columbia, MO 65211, USA.
Lett Appl Microbiol. 2002;35(6):508-12. doi: 10.1046/j.1472-765x.2002.01231.x.
To compare procedures for recovering template DNA from ground beef or chicken for polymerase chain reaction (PCR)-based detection of Salmonella.
The primer set of ST11 and ST15 was utilized to amplify a 429-bp product from Salmonella serotype Typhimurium. Boiling and three commercial kits were evaluated for extracting DNA from pure suspensions and artificially contaminated ground beef and chicken. The detection sensitivity of the PCR assay for pure cultures was independent of the template preparation method (P=0.946). Boiling and GeneReleaser failed to detect Salm. Typhimurium at 4 x 106 cfu g(-1) in ground chicken. PrepMan Ultra and the high pure PCR template preparation kit facilitated reliable and sensitive detection of Salm. Typhimurium in two types of food. The sensitivities were approx. 4 x 103 cfu g(-1). When spiked samples were enriched in peptone water for 6 h, an initial inoculum of 1 cfu g(-1) was detectable.
Four template DNA preparation methods differed in performance with respect to the type of samples tested.
Template DNA for the PCR detection of pathogenic bacteria, such as Salmonella in meat and poultry, could be effectively obtained using a simple rapid method such as the commercially available PrepMan Ultra kit.
比较从绞碎牛肉或鸡肉中回收模板DNA的方法，用于基于聚合酶链反应（PCR）检测沙门氏菌。
使用ST11和ST15引物对从鼠伤寒沙门氏菌中扩增出429 bp的产物。对煮沸法和三种商业试剂盒进行评估，以从纯菌悬液、人工污染的绞碎牛肉和鸡肉中提取DNA。PCR检测纯培养物的灵敏度与模板制备方法无关（P = 0.946）。煮沸法和GeneReleaser未能检测到鸡肉中4×10⁶ cfu g⁻¹的鼠伤寒沙门氏菌。PrepMan Ultra和高纯PCR模板制备试剂盒有助于可靠且灵敏地检测两种类型食品中的鼠伤寒沙门氏菌。灵敏度约为4×10³ cfu g⁻¹。当加标样品在蛋白胨水中富集6小时后，初始接种量为1 cfu g⁻¹时即可检测到。
四种模板DNA制备方法在测试样品类型方面的性能有所不同。
使用简单快速的方法，如市售的PrepMan Ultra试剂盒，可以有效地获得用于PCR检测肉类和家禽中病原菌（如沙门氏菌）的模板DNA。
