|Author(s)||Jérôme Marc1, Passerini Delphine1, Chevalier Frederique1, Marchand Laetitia1, Leroi Francoise1, Macé Sabrina1|
|Affiliation(s)||1 : IFREMER, BRM, EM3B Laboratory, F-44000 Nantes 3, France|
|Source||International Journal Of Food Microbiology (0168-1605) (Elsevier BV), 2022-02 , Vol. 363 , P. 109504 (8p.)|
|WOS© Times Cited||2|
|Keyword(s)||LAB, Seafood, Real-time PCR, TaqMan TM probe|
Quantification of lactic acid bacteria (LAB) is essential to control quality of seafood products like cold-smoked salmon (CSS). In the present study, we report the design and optimization of a dual-labelled TaqMan ™ probe targeting the V7 region of 16S rRNA gene for the detection of LAB in CSS. This quantitative PCR (qPCR) assays is useful for the simultaneous detection of the ten LAB genera communally encountered in CSS as Aerococcus, Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Macrococcus, Streptococcus, Vagococcus and Weissella. The specificity of this method was demonstrated against 14 genera (44 isolates, 35 species) of Gram-positive bacteria and 19 genera of Gram-negative (40 isolates, 34 species). Calibration of the method was performed in CSS matrix using a mix of equimolar cultured solution of five LAB. Quantification with the qPCR method range from 3.5 to 8.5 Log CFU/g in CSS matrix, covering 5 orders of magnitude. On these artificially contaminated CSS slices, PCR method results correlated successfully (R2 = 0.9945) with the conventional enumeration on Elliker medium. In addition, the new method was successful on commercial CSS from five different origins with a quantification range from 3.7 Log CFU/g to 8.0 Log CFU/g. This one-step quantitative methodology is proposed as a rapid and complementary tool of the cultural methods to investigate the LAB microbiota and biodiversity of CSS.
Jérôme Marc, Passerini Delphine, Chevalier Frederique, Marchand Laetitia, Leroi Francoise, Macé Sabrina (2022). Development of a rapid qPCR method to quantify lactic acid bacteria in cold-smoked salmon. International Journal Of Food Microbiology, 363, 109504 (8p.). Publisher's official version : https://doi.org/10.1016/j.ijfoodmicro.2021.109504 , Open Access version : https://archimer.ifremer.fr/doc/00741/85334/