Semiquantitative determination of Alicyclobacillus acidoterrestris in orange juice by reverse-transcriptase polymerase chain reaction and capillary electrophoresis--laser induced fluorescence using microchip technology.
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.
Funes-Huacca
Maribel
M
Instituto de Química de São Carlos, Universidade de São Paulo, Brazil.
Regitano
Luciana Correia de Almeida
LC
Mueller
Odilo
O
Carrilho
Emanuel
E
D001407:Bacillus / Q000302:isolation & purification*; D001628:Beverages / Q000382:microbiology*; D032084:Citrus sinensis / Q000382:microbiology*; D046209:Electrophoresis, Microchip / Q000379:methods*; D005453:Fluorescence; D005506:Food Contamination / Q000032:analysis; D007834:Lasers; D012329:RNA, Bacterial / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D012329:RNA, Bacterial