New molecular targets specific to Listeria and 6 common Listeria species and their rapid detection methods
A technology for Listeria and Listeria gasseri, applied in the field of identification of Listeria, Listeria and 6 common Listeria specific new molecular targets and its rapid detection, which can solve missing and increase false positive errors detection and false negative missed detection, mutations and other issues
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0047] Example 1 Mining of Listeria and 6 Common Listeria Specific New Molecular Targets
[0048] According to the GenBank database and the whole genome DNA sequence of Listeria self-tested by our team, bioinformatics analysis was carried out; Listeria and 6 common Listeria species (Listeria monocytogenes, Listeria innoctae, Specific gene fragments of Listeria ovis, Listeria welsii, Listeria westernis and Listeria gasseri), the nucleotide sequences of said gene fragments are as shown in SEQ ID NO.1-SEQ ID NO.16 shown.
[0049] Wherein, the sequence SEQ ID NO.1 is used to identify Listeria; the sequence SEQ ID NO.2 is used to identify Listeria monocytogenes; the sequence SEQ ID NO.3-5 is used to identify English Listeria knauci; said sequence SEQID NO.6-8 is used to identify Listeria ovis; said sequence SEQ ID NO.9-13 is used to identify Western-style Listeria; said sequence SEQ ID NO.14 is used for identifying Listeria welsii; said sequences SEQ ID NO.15 and 16 are used for ...
Embodiment 2
[0050] Example 2 Rapid Detection of Primer Effectiveness
[0051] 1) Primer design
[0052] According to the sequence SEQ ID NO.1-16 in Example 1, a specific PCR amplification primer set (including a forward primer and a reverse primer) was designed. The sequence of the primer set is shown in Table 1.
[0053] Table 1 Specific PCR detection primer set
[0054]
[0055]
[0056] 2) The method for identifying Listeria, the steps are as follows:
[0057] S1 DNA template preparation: the bacterial strains to be tested were enriched and cultured in LB liquid medium, and the bacterial genomic DNA was extracted using a commercial bacterial genomic DNA extraction kit as the template to be tested;
[0058] S2 PCR amplification: use one of the primer sets 1 to 16 to perform PCR amplification on the DNA of the sample to be tested;
[0059] ①PCR detection system:
[0060]
[0061]
[0062] in:
[0063] When the template DNA is used to identify whether there is Listeria, t...
Embodiment 3
[0074] The specificity evaluation of embodiment 3 Listeria PCR detection method
[0075] A total of 94 strains of Listeria monocytogenes, 2 strains of Listeria innocua, 1 strain of Listeria ovis, 1 strain of Western-style Listeria, 2 strains of Listeria welsii, and 1 strain of Listeria gasseri were collected. 101 strains of Listeria and 16 strains of non-Listeria (including Enterobacter sakazakii, Staphylococcus aureus, etc.) were detected by PCR according to the method in Example 2. Wherein, the S1 DNA template is prepared by extracting the genomic DNA of each bacterium respectively; during the S2 PCR amplification, the primers used are the primers in primer set 1.
[0076] The gel results of PCR amplification products are as follows: figure 1 , M is DL2000 DNA standard marker, 1-101 are Listeria strains, 102-117 are non-Listeria strains, C is negative control.
[0077] The strains and test results are shown in Table 2 below; in the table, "+" in the test result column indi...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More