关键词: <, p>, 环介导等温扩增技术, 金黄色葡萄球菌, 实验动物<, /p>

Abstract: Abstract: Objective To establish a sensitive and efficient method for detecting Staphylococcus aureus in laboratory animals． Method A set of four primers targeting the heat-stable nuclease ( nuc) gene( V01281. 1) of S． aureus were designed and the specifity of the Lamp assay was evaluated by common pathogenic bacteria in laboratory animals． The reaction conditions of LAMP were optimized including Mg2 + concentration、dNTPs concentration and time． Using LAMP and PCＲ method to detect S． aureus in pure culture and artificially contaminated feces and compared their sensitivity． Ｒesult No cross reaction was found in non-S． aureus bacteria and the optimal reaction condition of LAMP were 1. 6 μmol /L of each inner primer，0. 2 μmol /L of each outer primer，2. 0 mmol /L of each dNTP，8. 0 mmol /L Mg2 + and incubated at 60 ℃ for 50 min． In pure culture，the detection limit of LAMP was 9CFU /test( 9 × 103 CFU /mL) compared with 9 × 101 CFU /test( 9 × 104 CFU /mL) of PCＲ． In artificially contaminated feces，the detection limit of LAMP was 4. 49 × 104 CFU /0. 1 g feces compared with 4. 49 × 105 CFU /0. 1 g feces of PCＲ． Conclusion The developed LAMP method is simple、sensitive and specific，and is available for rapid detection of S． aureus in laboratory animals．

Key words: <p>LAMP, Staphylococcus aureus, laboratory animal</p>