41 results about "Circulating miRNA" patented technology

The invention relates to the field of biomedicine, in particular to a method for directly detecting real-time fluorescent quantitative RT-qPCR of circulating microRNA (miRNA) in a blood serum or blood plasma without the need of extracting nucleic acid. The method comprises the steps that S1, exosome and a miRNA protein complex in the blood serum or the blood plasma are pyrolyzed and centrifuged, and then a circulating miRNA crude extract is obtained; S2, the miRNA is subjected to tailing and reverse transcription; and S3, the RT-qPCR is quantitatively detected. According to the amplification method for the direct fluorescent quantitative RT-qPCR of the miRNA [Direct S-Poly(T)Plus, for short, DSPP], the nucleic acid does not need to be extracted, Poly(A) tailing and reverse transcription of the miRNA are synchronously completed in a reaction system, operation is easy and convenient, the time is shortened, and cDNA can be prepared within 95 minutes. Compared with a stem-loop method, the sensitivity of the technical system is increased by dozens of times and even hundreds of times, and the technical system which is used for miRNA detection and is simple, convenient, sensitive, efficient, rapid and inexpensive is built. The technical system is especially suitable for clinic application and popularization, and detection of the miRNA in a biological fluid sample with the low natural abundance of the miRNA.

The present invention discloses a circulating miRNA marker related to auxiliary diagnosis of papillary carcinoma of thyroid and application thereof. The marker is one or more selected from plasma markers miR-346, miR-10a-5p and miR-34a-5p and serum markers miR-25-3p, miR-296-5p and miR-92a-3p. As a novel biomarker, circulating miRNA has the characteristics of good stability, easy preparation through minimally invasive operation, high sensitivity and high specificity, and development and utilization of molecular markers will provide new directions for the diagnosis and further treatment of various diseases including tumors. The circulating miRNA marker having a clinical diagnosis potential for papillary carcinoma of thyroid is provided by the invention in a more targeted manner. Researchesconfirm reliability and reproducibility of this group of miRNAs as non-invasive markers for diagnosis of papillary carcinoma of thyroid.

Provided is a novel method for the early detection and/or discrimination of cancer and anti-cancer agents for the prevention or early treatment of cancer. In particular, the method relates to the determination of levels of circulating miRNAs in breast cancer patients, both primary and metastatic. Furthermore, kits, devices, pharmaceutical compositions as well as methods related thereto are described.

The invention relates to a miRNA expression model for diagnosing hepatic diseases independently. In the invention, an expression model of a miR-885-5p which is one of the circulating miRNAs associated with hepatic diseases is built by analyzing the types and relative expression levels of potential circulating miRNAs in a serum sample of a patient with cirrhosis or hepatocellular carcinoma and comparing the expression levels of the potential circulating miRNAs with the expression abundance of the circulating miRNAs in healthy reference serum. The model is formed by the expression indexes of the miR-885-5p, wherein the expression level of the miR-885-5p is higher than that of a healthy reference and is less tan 0.0001. ROC curve analysis shows that the hepatic disease HCC, LC and CHB identification efficacy AUC of the miR-885-5p and the healthy reference is 0.904, and the sensitivity and specificity of the miR-885-5p and the healthy reference are 90.5 percent and 79.2 percent respectively.

The invention relates to application of circulating miRNA in serving as age-related macular degeneration diagnostic markers. Expression differences of miRNA between patients with neovascular AMD and patients with cataract are detected through a miRNA expression chip, ten pieces of miRNA are selected from all miRNA subjected to differential expression for verification, a sample is enlarged, special miRNA expression is conducted through a fluorescent quantitative PCR method, comparison is conducted on the content of different miRNA in the patients with AMD and a control group, and the feasibility of serving as the diagnostic markers is verified. It is displayed through ROC curve analysis that AUC of miR-27a-3p, AUC of miR-29b-3p and AUC of miR-195-5p are 79.1%, 72.7% and 70.7% respectively. It is shown that miR-27a-3p, miR-29b-3p and miR-195-5p can serve as the age-related macular degeneration diagnostic markers.

The invention provides a microchip used for early diagnosis of the lung cancer and based on a semiconductor quantum dot, and a method of applying the microchip in preparation of a DNA probe used for detecting circulating miRNAs extracted from peripheral blood serum of a high risk group. The semiconductor quantum dot is used as a fluorescence detection signal of the microchip and is a multi-shell-structured Cd/Ge semiconductor quantum dot with a particle size of 3 to 6 nm; and the shell layer of the semiconductor quantum dot is coated by an inorganic layer selected from a group consisting of a CdS inorganic layer and a ZnS inorganic layer. According to the invention, directed at 7 to 38 circulating miRNAs in an early stage of the lung cancer, the method employs improved hybridization process and a DNA-quantum dot fluorescence signal to detect low-abundance miRNAs in serum, so a detection system applicable to clinical diagnosis and according with national in-vitro diagnostic reagent and quality control standards is formed; quantum dot nanometer technology and microchip technology are combined together, the miRNA detection system applicable to early serological diagnosis of the lung cancer is provided; and the detection system can distinguish benignancy and malignancy of lesion in the early stage of the lung cancer, enables patients with the lung cancer to be treated on time and prolongs the lifetime of the patients.

The invention discloses application of circulating nucleic acid serving as a breast cancer biomarker. Via a large number of literature researches, 96 miRNAs which have differential expression in a breast cancer tissue and body fluid or have functional verification for a level of a breast cancer cell line are screened out and used as candidate genes for researching; particularly, a prognosis relationship between the circulating miRNAs and long-term follow-up people suffering from the breast cancer is researched in the application; for 182 patients, based on bivariate regression analysis of machine training, 2 to 10 miRNA combinations have a distinguishing effect on prognosis of the breast cancer patients; multi-factor Cox regression analysis shows that a combination of 2 to 7 miRNAs is used as an independent risk factor of disease free survival and distant disease free survival of the breast cancer patients; particularly in lymph node-negative patients, a combination of 4 to 5 miRNAs has a better survival forecasting effect on the low and high-risk breast cancer patients. The research provides possibility for the application of the circulating miRNA in serving as a tumor marker.

The invention provides circulating miRNAs (namely microRNAs) for early diagnosis of acute coronary syndromes and an application thereof. The circulating miRNAs comprise miR-3149 and/or miR-122, further comprise miR-2861, and further comprise miR-140-3p and/or miR-720. The circulating miRNAs provided by the invention can be taken as markers to be applied to early diagnosis of the acute coronary syndromes.

A diagnostic kit to detect lung adenocarcinoma, or to stratify patients according to expected prognosis comprising at least one oligonucleotide probe capable of binding to at least a portion of a circulating miRNA selected from the group comprising miR-556, -550, -939, -616*, -146b-3p, -30c-1*, -339-5p and -656.

The invention discloses a circulating miRNA marker and a carcino-embryonic miRNA marker related to pan-tumor auxiliary diagnosis and application of the circulating miRNA marker and the carcino-embryonic miRNA marker. The circulating miRNA marker and the carcino-embryonic miRNA marker are taken as novel biomarkers and have the characteristics of being good in stability, minimally invasive, easy toobtain and high in sensitivity and specificity. Development and utilization of the molecular markers provide a new direction for diagnosis and further treatment of various diseases including tumors. According to the invention, the generic tumor circulating miRNA marker and the carcino-embryonic miRNA marker with clinical diagnosis potential can be obtained in a more targeted manner. Researches prove the reliability and repeatability of the group of miRNAs as noninvasive markers for diagnosing pan-tumors.

The invention provides a visual detection method and a detection probe for simultaneously detecting two circulating miRNAs. The method comprises the following steps: step 1, preparing gold nanoparticles; step 2, preparing a gold nanoparticle-substrate DNA chain HP compound HP (at) AuNPs, and combining the compound HP (at) AuNPs with enzyme chains A and B to form an AuNPs-DNA walker detection probe; step 3, performing signal amplification reaction on the target miRNA and a detection probe; and step 4, carrying out visual detection on the target miRNA based on chemiluminescence. The detection probe provided by the invention can realize simultaneous quantitative detection of a plurality of samples containing two different circulating miRNAs, and detection results are not interfered with each other. According to the detection method provided by the invention, a chemiluminescence signal is recorded in a mobile phone photographing manner, expensive instruments are not needed, the cost is reduced, the detection operation is simplified, and immediate detection of miRNA can be realized.

The invention relates to a method for detecting the expression level of circulating miRNA-4741, and an application of the miRNA-4741 in the diagnosis and prognosis of a liver cancer patient. The miRNA-4741 having a high expression level, used as the target marker of experiments, is found through carrying out high-throughput screening on the kind of the circulating miRNA in the blood plasma of a patient diagnosed with the liver cancer and comparing the miRNA with corresponding miRNA in the blood plasma of a healthy control. The expression level of the miRNA-4741 is higher than that of a controlgroup, and P is 0.001. An ROC curve analysis result shows that the usefulness AUC of the miRNA-4741 in the identification of normal people and a liver cancer patient is 0.765, the sensitivity is 75.4%, and the specificity is 66.7%. A survival curve analysis result shows that the life is short if the expression level is high, and the life is long if the expression level is low.

According to the invention, small RNA sequencing is carried out on serum and blood cells of patients with coronary heart diseases respectively, and based on parameters such as difference change of miRNA and serum abundance in sequencing data and detection of methodological stability, applicable specific candidate miRNA markers are systematically screened out. Furthermore, through cooperation witha prevention and treatment professional team for cardiovascular populations, researches are conducted on application of the candidate miRNA markers to diagnosis and prediction of atherosclerosis-related diseases during large-scale follow-up visit of the population. Research results in the invention provide important theoretical and methodological basis for clinical application of circulating miRNA.

The invention relates to an application of circulating miRNA-4731-3p as a diagnostic marker for primary lung cancer, and method for detecting the miRNA-4731-3p. The miRNA-4731-3p having a high expression level, used as the target marker of experiments, is found through carrying out high-throughput screening on the kind of the circulating miRNA in the blood plasma of a patient diagnosed with the lung cancer and comparing the miRNA with corresponding miRNA in the blood plasma of a healthy control. The expression level of the miRNA-4731-3p is obviously higher than that of the control, and P is less than 0.001. An ROC curve analysis result shows that the usefulness AUC of the miRNA-4731-3p in the identification of normal people and a lung cancer patient is 0.048, the sensitivity is 95.0%, andthe specificity is 57.5%. A survival curve analysis result shows that the life is short if the expression level is high, and the life is long if the expression level is low.