Microflora marker for predicting gastric cancer lymph node metastasis and application thereof
A technology for lymph node metastasis and markers, which is used in applications, medical preparations containing active ingredients, microorganisms, etc., to achieve the effect of good detection specificity and high sensitivity
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Embodiment 1
[0028] Example 1. Micro ecological analysis of gastric cancer tissue
[0029] (1) Gastric cancer tissue microorganisms DNA extraction:
[0030] During the surgery of gastric cancer patients, the surgical doctor will separate the gastric specimens, properly clean the gastric specimen, and use sterile equipment to dissect it.0.1 % ethylene pyrogenesis iodine disinfecting the surface was cleaned and disinfected, and then the frozen storage tube was loaded, and the liquid nitrogen was transferred to transfer.
[0031] The thawing gastric cancer tissue sample was fully grinded using the grinding pearl average (BIOSPEC).
[0032] (2) Swalm of gastric cancer tissue microorganisms 16S RRNA amplification subquitm:
[0033] The extracted gastric cancer tissue microorganisms DNA use 341F 5′-CCTACGGGGCWGCWGCAG-3 ′ (SEQ ID NO: 1) and 806r 5′-GGACTACHVGGTWTAAT-3 ′ (SEQ ID NO: 2), and use UCPMultiplex PCR Kit (qiagenn) The 16S RRNA gene V3-V4 area amplification is made, and the Qiasequltralow I...
Embodiment 2
[0037] Example 2. Screening the characteristic gastric flora of patients with gastric cancer lymph nodes
[0038] (1) Establishment of the research queue
[0039]In this study, 9 cases of gastric cancer patients were collected. The primary stove tissue samples were performed. After surgery, all patients were diagnosed to determine whether there were lymph node metastasis. Four patients had no lymph nodes and 5 patients had lymph nodes metastasis.
[0040] (2) Features of gastric cancer tissue groups of gastric cancer lymph node metastasis
[0041] Micro ecological analysis of gastric cancer tissue groups is performed according to the method of Example 1.Based on gastric cancer patients with no lymph nodes metastasis, linear discrimination analysis effect (Linear Discriminant Analysis EffectSize (Lefse) method analyzes the characteristic flora of gastric cancer lymphoskin metastasis.
[0042] First of all, use non-parameter test KRUSKAL-WALLIS rank and test to identify the abundan...
Embodiment 3
[0044] Examples 3. Evaluation of gastric cancer tissue microbial markers predict the diagnostic effectiveness evaluation of gastric cancer lymph nodes metastasis experiment
[0045] (1) Revolution and testing of gastric cancer patients verifying the queue
[0046] A sample of primary stove tissue was collected in accordance with the scheme of the embodiment 2, 4 of which had lymph nodes metastasis and 4 cases of no lymph nodes were transferred.The micro -ecological testing scheme of the gastric cancer tissue is based on Example 1.
[0047] (2) Evaluation of the diagnostic effectiveness of gastric cancer tissue microbial tags predict gastric cancer lymph nodes metastasis.
[0048] The test results reminded that the relative abundance of 4 patients with lymph nodes metastasis gastric cancer patients (Enterobacteriales) Yersinia (2.250 % vs 0.133 %, which is significantly higher than that of Yersinia.P = 0.021).The Receiver OperationCharacteristic Curve (ROC) found that the relativ...
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