[Show abstract][Hide abstract] ABSTRACT: Objective:
To evaluate the adsorption capacity of magnetic beads for common foodborne pathogens.
Escherichia coli, Staphylococcus aureus, Listeria monocytogenes and Cronobacter sakazakii were used as targets, the suitable conditions were obtained by optimization the incubation time, bead size, reaction temperature and so on. The adsorption efficiency of the magnetic beads in analog samples was also tested.
The optimum conditions were as follows: 2.0 - 3.0 microm diameter magnetic beads 50 microl, incubation for 60 min under 37 degrees C. In analog samples, the adsorption efficiency of the magnetic beads for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes and Cronobacter sakazakii in juice, milk, duck meat were 50% - 84%, 46% - 66%, 54% - 74%, respectively.
Magnetic beads enrichment technique has high adsorption efficiency, simple to operate, which is meet the requirement for PCR detection.
No preview · Article · Jul 2014 · Wei sheng yan jiu = Journal of hygiene research
[Show abstract][Hide abstract] ABSTRACT: A TaqMan probe real-time PCR method was developed for rapid detection of yak component in raw and cooked meat products. Specific primers and TaqMan probes of yak (Bos grunniens) were designed in the cytochrome b gene. The specificity of the method was evaluated using pure meat of eight yak breeds (Jiulong, Qinghai plateau, Maiwa, Gannan, Bazhou, Sibu, Zhongdian, and Jiali) samples and nine non-Bos grunniens animals (sheep, goat, pig, chicken, cattle, water buffalo, donkey, horse, and rabbit). DNA showed no cross-reaction with non-Bos grunniens animal DNA. This method proved to be sensitive in detecting the presence of low levels of target DNA obtained from 0.001% (w/w) component in a mixed meat sample. The method also successfully identified commercial yak meat products. The results showed that some yak meat might be involved in business fraud by using cattle meat (in this paper, cattle meat means meat of Bos taurus) instead of yak meat. In conclusion, real-time PCR assay used in this study was shown to be a rapid and sensitive method for detection of yak DNA in fresh meat and cooked meat products.
No preview · Article · Mar 2013 · Journal of AOAC International
[Show abstract][Hide abstract] ABSTRACT: A total of 48 Listeria monocytogenes isolates of different import food products from 8 provinces between 2005 and 2008 were characterized. The serotype and virulence were confirmed for each strain and molecular subtyping were analyzed by multilocus sequence typing (MLST). Twenty five strains were assigned to serotype 1/2a, and 11 isolates to serotype 1/2b, serotype 4b were found in 7 isolate, and the remaining 5 strains were grouped into serotypes 1/2c, 4a, and 4e. Molecular subtyping schemes found thirty two sequence types (STs) among these isolates and the majority of L. monocytogenes strains belonged to lineage II (56%), followed by lineage I (38%), lineage III (6%). Two molecular subtype clusters, cluster A included all isolates of lineage II, while cluster B contained the isolates of lineages I and lineages III. Two L. monocytogenes strains were not grouped in either of the two clusters. Fifty three isolates were as virulent as L. monocytogenes reference strain EGD in mouse virulence assay, while the isolates 22213 and 22265 had low pathogenicity. These results provide the first molecular insight into the L. monocytogenes strains isolated from import food products of 8 provinces in China and indicate the potential risk to cause human disease if intake by contaminated foods. MLST could be used as a routine subtyping method of L. monocytogenes isolates. In China, inspection and quarantine strategies of imported foods should be strengthened. PRACTICAL APPLICATION: There is a potential risk of listeriosis in China and routine subtyping of L. monocytogenes isolates is important. It is necessary for food hygiene management to strengthen the supervision of imported foods.
No preview · Article · Feb 2012 · Journal of Food Science
[Show abstract][Hide abstract] ABSTRACT: To establish a real-time PCR assay for the rapid detection of Listeria monocytogenes in simulated milk specimens.
Based on part fragments of hlyO gene, a pair of primers and Taq-Man probe were designed for quantitative detection of L. monocytogenes. The specificity of the primers and probe were tested by using different L. monocytogenes strains and other common pathogenic bacteria.
L. monocytogenes strains were positive in the detection and other tested strains were negative. The sensitivity of assay was 9 copies per PCR reaction.
The specificity and sensitivity of Taq Man real-time PCR technology for detecting L. monocytogenes in simulated dairy specimens were high, and the assay could be completed within 1.5 h. This method could be used to detect other food samples contaminated by L. monocytogenes and identify the cause of food-borne Listeriosis outbreaks.
No preview · Article · Nov 2011 · Wei sheng yan jiu = Journal of hygiene research