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piRNA (piwi-interacting ribonucleic acid) biomarkers for noninvasive early-stage diagnosis on colon cancer and rectal cancer and detection kit

A biomarker, early diagnosis technology, applied in the determination/test of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc. The effect of saving operation time

Pending Publication Date: 2020-05-29
内蒙古医科大学附属人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, serum tumor markers for colon and rectal cancer commonly used in clinical practice, such as CEA and CA-199, do not increase in the serum of many early-stage patients, and cannot meet the needs of early screening. Currently, they are mainly used for curative effect observation and monitoring. recurrence monitoring

Method used

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  • piRNA (piwi-interacting ribonucleic acid) biomarkers for noninvasive early-stage diagnosis on colon cancer and rectal cancer and detection kit
  • piRNA (piwi-interacting ribonucleic acid) biomarkers for noninvasive early-stage diagnosis on colon cancer and rectal cancer and detection kit
  • piRNA (piwi-interacting ribonucleic acid) biomarkers for noninvasive early-stage diagnosis on colon cancer and rectal cancer and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Preliminary screening of abnormally expressed piRNAs in the serum of patients with colon and rectal cancer

[0023] 1. Collection of serum samples

[0024] A total of 86 patients with early colon and rectal cancer (stage 0-II) diagnosed between July 2015 and May 2018 in the Affiliated Hospital of Inner Mongolia Medical University were selected as the patient group, and 66 healthy individuals confirmed by physical examination were selected as the control group. All patients with colon and rectal cancer were diagnosed for the first time, and had not undergone surgery, radiotherapy or chemotherapy before blood collection. Individuals in the control group did not suffer from malignant tumors or other diseases, and were matched with the individuals in the patient group in terms of age and sex.

[0025] In the morning on an empty stomach, draw 5 ml of peripheral blood into a coagulation blood collection tube, let it stand for 30 minutes, centrifuge at 4 ℃, 3900rmp...

Embodiment 2

[0028] Example 2: Verification of differentially expressed serum piRNA and its value as a diagnostic marker for colon and rectal cancer

[0029] 1. RNA extraction

[0030] Serum samples from 80 patients and 60 age- and gender-matched healthy people were collected, and serum total RNA was extracted by the TRIzol method: for every 200 μL serum, add 1 mL TRIzol, vortex and mix, and place at room temperature for 5 min; add 0.2 mL chloroform, Vigorously shake for 15 s, let stand for 3 minutes; centrifuge at 4°C, 10,000 rpm / min for 10 minutes, transfer the upper aqueous phase to a new tube; add an equal volume of isopropanol, mix well, let stand for 20 minutes; 4°C, Centrifuge at 10,000 rpm / min for 10 min, remove the supernatant; wash the pellet with 1 mL of 75% ethanol; centrifuge at 5,000 r / min at 4°C for 5 min, discard the liquid; after drying at room temperature, add 20 μL of enzyme-free water to dissolve the RNA , Take 2 μL of RNA and measure the RNA concentration and A260 / 280...

Embodiment 3

[0039] Embodiment 3: receiver operating characteristic curve (ROC) analysis

[0040] A ROC curve was constructed to analyze the diagnostic ability of two serum piRNAs to distinguish early colon and rectal cancer patients from healthy people. The area under the curve of piRNA020619 was 0.883 (95% confidence interval: 0.822-0.944), and at the best cutoff value, its sensitivity and specificity were 80.6% and 82.4%, respectively. The area under the curve of piRNA020450 was 0.774 (95% confidence interval: 0.694-0.855); at the best cutoff value, its sensitivity and specificity were 71.2% and 74.0%, respectively.

[0041] Further, the relative expression of these two piRNAs in serum was introduced into the logistic regression equation, and the logit (P) discriminant formula was established: logit (P)=-3.332+0.703×piRNA020619+0.327×piRNA020450. Based on the verification of the discriminant formula, it was found that the combined use of piRNA020619 and piRNA020450 has high diagnostic ...

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Abstract

The invention discloses piRNA (piwi-interacting ribonucleic acid) biomarkers for noninvasive early-stage diagnosis on colon cancer and rectal cancer and a detection kit. The piRNA biomarkers comprisepiRNA020619 and piRNA020450. The expression amounts of the two piRNA biomarkers in serum of patients suffering from colon cancer and rectal cancer are remarkably higher than those in serum of healthypeople, and the piRNA biomarkers have very high sensitivity and specificity when being adopted to diagnose colon cancer and rectal cancer. By adopting the kit disclosed by the invention, serum samplesof patients suffering from colon cancer and rectal cancer and serum samples of healthy people can be rapidly and accurately distinguished.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a piRNA biomarker for the diagnosis of human colon and rectal cancer and a detection kit based on the marker. Background technique [0002] Colon and rectal cancer is one of the most common clinical malignant tumors. There are more than 1.4 million new cases and more than 700,000 deaths worldwide each year. In my country, the morbidity and mortality of colon and rectal cancer are increasing year by year, with more than 250,000 new cases and more than 140,000 deaths each year, which seriously threatens the health and life of the people and has become an urgent public health problem. [0003] The prognosis of patients with colon and rectal cancer is closely related to tumor stage. The 5-year survival rate of early stage patients is more than 80%, while the 5-year survival rate of advanced stage patients is less than 20%. Unfortunately, the onset of colon and rectal cancer is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 王振飞贾永峰马大光郝钦冯立王玲杨永雁陈永霞张慕尧谭晓慧
Owner 内蒙古医科大学附属人民医院
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