| Autor: |
SUN Xueqi, YIN Weilu, JIANG Jiaxi, ZHANG Mingming, LIANG Meidan, ZENG Shanmei, XIAO Jian |
| Jazyk: |
čínština |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
Zhongguo shipin weisheng zazhi, Vol 35, Iss 11, Pp 1571-1578 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
1004-8456 |
| DOI: |
10.13590/j.cjfh.2023.11.004 |
| Popis: |
ObjectiveThis study aimed to establish a quantitative 3-plex droplet digital polymerase chain reaction （ddPCR） method for simultaneously detecting the copy numbers of Salmonella， Bacillus cereus， and Listeria monocytogenes in instant food.MethodsThree pairs of primers and probes corresponding to three single-copy-genes were selected as target genes. The genes were the essC gene in Bacillus cereus， ttrA/ttrC gene in Salmonella， and invasion-associated endopeptidase gene in Listeria monocytogenes. The specificity of the primers and probes were verified by real-time fluorescence quantitative PCR separately. A 3-plex ddPCR method was constructed to detect the copy numbers of three pathogenic bacteria simultaneously.ResultsThe linear ranges were： 25-22 687 copies/20 µL for Salmonella， 19-15 620 copies/20 µL for Bacillus cereus， and 18-23 373 copies/20 µL for Listeria monocytogenes. The three linear correlation coefficients were r≥0.999. relative standard deviation （RSD）≤12% at six concentrations and repeated thrice， indicating favorable repeatability. The minimum detection limits were six copies/20 µL for Salmonella， three copies/20 µL for Bacillus cereus， and seven copies/20 µL for Listeria monocytogenes. When a simulated sample of contaminated rice noodles was detected by 3-plex ddPCR and the plate counting method， the deviation between these two methods was |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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