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Early-stage cancer screening kit based on blood plasma exosome GRP78 mRNA

A kit and exosome technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., to achieve the effect of good specificity, high reliability, and easy clinical promotion

Inactive Publication Date: 2018-05-08
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in practical application work, there is no kit for early cancer screening using exosomes. The determination and selection of specific markers in exosomes is one of the most important key factors. At present, there is no suitable exosome Body markers for early detection of cancer

Method used

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  • Early-stage cancer screening kit based on blood plasma exosome GRP78 mRNA
  • Early-stage cancer screening kit based on blood plasma exosome GRP78 mRNA
  • Early-stage cancer screening kit based on blood plasma exosome GRP78 mRNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A cancer early screening kit based on GRP78 mRNA in plasma exosomes, including exosomal GRP78 mRNA fluorescence quantitative PCR upstream primers and PCR downstream primers as shown in SEQ ID NO: 1-2, EDTA anticoagulant tube, DNaseI , TRIZOL, PrimeScript TM 1st Strand cDNA Synthesis Kit, Premix Ex Taq TM , negative control substance and positive standard substance, wherein the negative control substance is pGEM-7Zf(+) empty vector, and the positive standard substance is pGEM-7Zf(+) plasmid vector containing inserted GRP78 specific nucleic acid sequence.

[0035] 1. The preparation process of the positive standard includes the following steps:

[0036] (1) PCR reaction

[0037] Because the abundance of mRNA in human peripheral blood leukocytes is high, GRP78 (NCBIReference Sequence: NM_005347.4) of mRNA in human peripheral blood leukocytes is used as a template, and reverse-transcribed into cDNA, PCR forward primer: as shown in SEQ ID NO: 4 Reverse primer: AAGCTCTC...

Embodiment 2

[0064] 1. According to the method of Example 1, the peripheral blood samples of healthy adults and patients with primary liver cancer, colorectal cancer, gastric cancer, non-small cell lung cancer, glioma, breast cancer and prostate cancer were detected, and the specific sample information is shown in Table 1 below shown. Provides standard cDNA concentrations of

[0065] Table 1

[0066]

[0067]

[0068] 2. The test results are shown in Table 2:

[0069] Table 2

[0070]

[0071] Note: SD of healthy adult group=12.70 copies / mL

[0072] The mean copy numbers of GRP78 mRNA in exosomes were 28.22 copies / mL (range 0-116) in the healthy adult group, 28.91 copies / mL (range 10-45) in the female group, and 27.90 copies / mL (range 10-45) in the male group. 0-116); hepatocellular carcinoma group 285.63 copies / mL (range 57-582), colorectal cancer group 117.82 copies / mL (range 27-347), gastric cancer group 107.88 copies / mL (range 40-575), non-small cell The lung cancer group...

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Abstract

The invention discloses an early-stage cancer screening kit based on blood plasma exosome GRP78 mRNA. The early-stage cancer screening kit comprises a pair of exosome GRP78 mRNA fluorogenic quantitative PCR primers, and the nucleotide sequences of the primers are shown in SEQ ID NO:1-2. By adopting the kit, early-stage screening of multiple types of cancer can be completed with only 5ml of blood,and an early-stage cancer screening method is good in specificity, high in sensitivity, high in efficiency, good in practicability, good in noninvasiveness and high in reliability, and easy in clinical popularization in medical institutes of different grades, is particularly applicable to physical screening and early-stage warming of susceptible tumor patients, and has great significant values.

Description

technical field [0001] The present invention relates to the technical field of lung cancer screening, and more specifically, to an early cancer screening kit based on plasma exosome GRP78 mRNA. Background technique [0002] Cancer refers to malignant tumors originating from epithelial tissue, and is the most common type of malignant tumors. Compared with benign tumors, malignant tumors grow faster and show invasive growth, are prone to bleeding, necrosis, ulcers, etc., and often have distant metastasis, resulting in emaciation, weakness, anemia, loss of appetite, fever, and severe organ damage. Impairment of function, etc., eventually lead to death of the patient. [0003] A large number of patients die from cancer every year. It is estimated that in the next 25 years, 300 million people in the world will suffer from cancer, of which 100 million will die from this disease. Cancer will become the number one killer of human beings in the new century and become one of the big...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 朱萧罗辉丁红荣
Owner GUANGDONG MEDICAL UNIV
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