Rapid, specific, and sensitive detection of spoilage molds in orange juice using a real-time Taqman PCR assay.
The outgrowth of spoilage organisms, including molds and yeasts, results in significant financial loss to the food industry and wastes natural resources. The objective of this study was to develop a rapid, specific, and sensitive real-time PCR method for detecting spoilage molds during screening of raw materials and final product quality control analysis. The 18S rRNA gene was used to develop PCR primers and probe. With this set of primers and probe, less than 1,000 mold cells per milliliter of orange juice (10 cells per reaction) were detected with the real-time PCR system within 6 to 7 h. No cross-reactivity was found with other common foodborne bacteria, yeasts, or food ingredients. This technique is significantly faster than current detection and identification procedures, which take from days to weeks.
Wan
Kai
K
Department of Food Science and Technology, Ohio State University, Columbus 43210, USA.
Yousef
Ahmed E
AE
Schwartz
Steve J
SJ
Wang
Hua H
HH
D001483:Base Sequence; D001628:Beverages / Q000382:microbiology*; D032084:Citrus sinensis / Q000382:microbiology; D003429:Cross Reactions; D004271:DNA, Fungal / Q000032:analysis; D005506:Food Contamination / Q000032:analysis*; D005658:Fungi / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011786:Quality Control; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity; D013997:Time Factors; D015003:Yeasts / Q000302:isolation & purification*
D004271:DNA, Fungal; D012337:RNA, Ribosomal, 18S