TY - JOUR
T1 - Rapid and simple estimation of microbiological quality of raw milk using chromogenic Limulus amoebocyte lysate endpoint assay
AU - Rhee, Min Suk
AU - Kang, Dong Hyun
PY - 2002/9
Y1 - 2002/9
N2 - A rapid chromogenic Limulus amoebocyte lysate (LAL) endpoint assay for the enumeration of total mesophilic microbial loads and coliforms was investigated as a means to assess the microbiological quality of raw milk. For experiment 1, raw milk samples (n = 25) were stored in a refrigerator (2 ± 2°C) and then analyzed at regular intervals (1, 5, 10, and 15 days). For experiment 2, fresh raw milk samples (n = 50) were tested to determine the utility of the LAL assay for fresh raw milk. The sample was diluted threefold in a 96-well microtiter plate with pyrogen-free water and assayed with a chromogenic LAL kit to find a final reaction point. The LAL results were compared with standard plate counts (SPC) and coliform counts determined by conventional plating methods. The results of the LAL assay were strongly correlated to conventional SPC (r2 = 0.93; n = 100) and were highly correlated to coliforms (r2 = 0.74; n = 100). A highly significant linear relationship (r2 = 0.82; n = 50) was also observed between the predicted SPC based on the LAL value and the actual SPC. The results of LAL testing were classified into one of seven contamination groups. The data set for SPC was effectively differentiated using the LAL technique (P < 0.01). The chromogenic LAL assay was found to be a rapid (within 16 min) and simple (not requiring specific instruments) method for monitoring microbial levels in raw milk. This method may be successfully implemented to rapidly determine highly microbial contaminated raw milk (>3.0 log10 CFU/ml of SPC).
AB - A rapid chromogenic Limulus amoebocyte lysate (LAL) endpoint assay for the enumeration of total mesophilic microbial loads and coliforms was investigated as a means to assess the microbiological quality of raw milk. For experiment 1, raw milk samples (n = 25) were stored in a refrigerator (2 ± 2°C) and then analyzed at regular intervals (1, 5, 10, and 15 days). For experiment 2, fresh raw milk samples (n = 50) were tested to determine the utility of the LAL assay for fresh raw milk. The sample was diluted threefold in a 96-well microtiter plate with pyrogen-free water and assayed with a chromogenic LAL kit to find a final reaction point. The LAL results were compared with standard plate counts (SPC) and coliform counts determined by conventional plating methods. The results of the LAL assay were strongly correlated to conventional SPC (r2 = 0.93; n = 100) and were highly correlated to coliforms (r2 = 0.74; n = 100). A highly significant linear relationship (r2 = 0.82; n = 50) was also observed between the predicted SPC based on the LAL value and the actual SPC. The results of LAL testing were classified into one of seven contamination groups. The data set for SPC was effectively differentiated using the LAL technique (P < 0.01). The chromogenic LAL assay was found to be a rapid (within 16 min) and simple (not requiring specific instruments) method for monitoring microbial levels in raw milk. This method may be successfully implemented to rapidly determine highly microbial contaminated raw milk (>3.0 log10 CFU/ml of SPC).
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U2 - 10.4315/0362-028X-65.9.1447
DO - 10.4315/0362-028X-65.9.1447
M3 - Article
C2 - 12233856
AN - SCOPUS:0036715279
VL - 65
SP - 1447
EP - 1451
JO - Journal of Food Protection
JF - Journal of Food Protection
SN - 0362-028X
IS - 9
ER -