Funes-Huacca Maribel, Regitano Luciana Correia de Almeida, Mueller Odilo, Carrilho Emanuel
Instituto de Química de São Carlos, Universidade de São Paulo, Brazil.
Electrophoresis. 2004 Nov;25(21-22):3860-4. doi: 10.1002/elps.200406105.
The semiquantitative detection of Alicyclobacillus acidoterrrestris in orange juice by reverse-transcriptase polymerase chain reaction (RT-PCR) with a linear dynamic range of 2 x 10(5)-2 colony forming units (CFU)/mL in terms of cell count is described. Separation, detection, and quantification of the RT-PCR products were accomplished using the Agilent 2100 bioanalyzer in conjunction with the DNA 1000 LabChip kit. After 0 and 12 h of enrichment, it was possible to generate a linear standard curve between the amount of cells and amplicon concentration of RT-PCR and PCR products. Using this method, cell diminution was verified in samples of orange juice treated with a natural inhibitor (Sapindus saponaria), determining the persistence of viable cells. Semiquantitative RT-PCR using the Agilent 2100 bioanalyzer is a potentially useful approach for rapid in vitro determination of A. acidoterrestris and monitoring of inhibitor susceptibility for the orange juice-producing industry.
描述了通过逆转录聚合酶链反应（RT-PCR）对橙汁中嗜酸耐热脂肪芽孢杆菌进行半定量检测的方法，其细胞计数的线性动态范围为2×10⁵ - 2菌落形成单位（CFU）/mL。使用安捷伦2100生物分析仪结合DNA 1000 LabChip试剂盒完成RT-PCR产物的分离、检测和定量。在富集0小时和12小时后，能够在细胞数量与RT-PCR和PCR产物的扩增子浓度之间生成线性标准曲线。使用该方法，在经天然抑制剂（无患子）处理的橙汁样品中验证了细胞减少情况，确定了活细胞的持久性。使用安捷伦2100生物分析仪进行半定量RT-PCR是一种潜在有用的方法，可用于快速体外测定嗜酸耐热脂肪芽孢杆菌以及监测橙汁生产行业中抑制剂的敏感性。
