[Application and evaluation of loop-mediated isothermal amplification method for detecting of Listeria monocytogenes in food].
OBJECTIVE
The loop-mediated isothermal amplification (LAMP) detection method was applied to detect Listeria monocytogenes in food. The specificity and sensitivity of this method were evaluated through comparing it with Real-time PCR and conventional detection method.


METHODS
The LAMP primers were designed based on hly gene of Listeria monocytogenes. The LAMP method was applied to detect 88 Listeria monocytogenes, 1 reference strain ATCC 15313 of Listeria monocytogenes and 33 non-targets bacteria strains; base-material addition test and practical food samples detection were also conducted. Both of Real-time PCR and ISO 11290-1 methods were used as parallel detection method in addition to LAMP. The three kinds of methods were compared by specificity, sensitivity, detection limit and the detection result of practical food samples.


RESULTS
Both detection results of LAMP and Real-time PCR for 89 Listeria monocytogenes were positive (100%, 89/89), 33 non-targets bacteria strains were negative (100%, 33/33). The sensitivity of LAMP was 2 × 10² CFU/ml, which was consistent with Real-time PCR method (2 × 10² CFU/ml) and better than ISO 11290-1 method (2 × 10² CFU/ml). Base-material addition test result showed that the detection limit of the three kinds of methods were 3 CFU/25 g samples. And the result of practical food samples displayed the same detection rate of 4% in the three methods (2/45).


CONCLUSIONS
The LAMP method of Listeria monocytogenes established in this study has good specificity and sensitivity, which can be applied to the rapid detection of Listeria monocytogenes.
Microbiology Laboratory of Inspection and Quarantine Technology Center, Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing 100026, China.
Lei
L
Zhang
Jing
J
Zeng
Dan
D
Ma
Jinxia
J
Cheng
Email:zhysph@sina.com.
Haiyu
H
Zhang
D005506:Food Contamination / Q000032:analysis*; D005513:Food Inspection / Q000379:methods*; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*