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Micro ribonucleic acid (RNA) molecular mark for diagnosing gestational diabetes and detection kit thereof

A technology of gestational diabetes and kits, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc.

Inactive Publication Date: 2014-11-05
周玲 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the diagnosis of gestational diabetes can only rely on the "glucose tolerance" test. This test method needs to draw venous blood from pregnant women four times in a row, which brings some pain to pregnant women.

Method used

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  • Micro ribonucleic acid (RNA) molecular mark for diagnosing gestational diabetes and detection kit thereof
  • Micro ribonucleic acid (RNA) molecular mark for diagnosing gestational diabetes and detection kit thereof
  • Micro ribonucleic acid (RNA) molecular mark for diagnosing gestational diabetes and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1. Microarray analysis of miRNA expression profile changes in the placenta of pregnant women with gestational diabetes.

[0016] Take 20 placentas from patients with gestational diabetes and 20 placentas from normal pregnant women as controls. Tissue RNA was extracted as follows.

[0017] ①Take fresh placenta samples, mix 20 samples, freeze in liquid nitrogen, grind with a mortar until the tissue becomes powdery (use liquid nitrogen to keep the sample at a low temperature during the grinding process), add an appropriate amount of sample powder to 1ml of TRIZOL, and store at room temperature Leave it for 1h.

[0018] ②Add 200 μl of chloroform to TRIZOL, shake vigorously, let stand for 2 minutes, centrifuge at 4°C, 12000 rpm for 15 minutes.

[0019] ③ Remove the supernatant, add an equal volume of isopropanol to the supernatant, shake vigorously, let stand at room temperature for 10 minutes, centrifuge at 12000 rpm at 4°C for 10 minutes.

[0020] ④ Discard the ...

Embodiment 2

[0025] Embodiment 2 uses real-time PCR technology to verify the result of the chip

[0026] Take 18 placentas from patients with gestational diabetes and 18 placentas from normal pregnant women as controls. Tissue RNA was extracted as follows.

[0027] ① Take a fresh placenta sample, freeze it in liquid nitrogen, grind it with a mortar until the tissue becomes powdery (use liquid nitrogen to keep the sample at a low temperature during the grinding process), add an appropriate amount of sample powder to 1 ml of TRIZOL, and place it at room temperature for 1 hour.

[0028] ②Add 200 μl of chloroform to TRIZOL, shake vigorously, let stand for 2 minutes, centrifuge at 4°C, 12000 rpm for 15 minutes.

[0029] ③ Remove the supernatant, add an equal volume of isopropanol to the supernatant, shake vigorously, let stand at room temperature for 10 minutes, centrifuge at 12000 rpm at 4°C for 10 minutes.

[0030] ④ Discard the supernatant, and place the precipitate in a ventilated place t...

Embodiment 3

[0040] Example 3 Using real-time PCR technology to detect the expression level of miR-508 in the serum of pregnant women

[0041] The serum of 18 cases of gestational diabetes patients and 18 cases of normal pregnant women were taken as controls. RNA was extracted as follows.

[0042] ①Take 500ul of fresh serum sample, add 500ul of TRIZOL to the serum, shake vigorously for 30s, and place at room temperature for 5min.

[0043] ②Add 200ul of isopropanol to TRIZOL, invert and mix well, shake vigorously for 2 minutes until the liquid is transparent, and let stand at room temperature for 5 minutes.

[0044] ③ 4°C, 12000rpm, centrifuge for 10min, and transfer the supernatant to a new eppendorf tube.

[0045] ④Add 500ul chloroform to the supernatant, mix by inverting, shake vigorously for 1min, and let stand at room temperature for 5min.

[0046] ⑤ 4°C, 12000rpm, centrifuge for 10min, and transfer the supernatant to a new eppendorf tube.

[0047] ⑥ Add 0.75 times the volume of is...

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Abstract

The invention provides a micro ribonucleic acid (RNA) molecular mark for diagnosing gestational diabetes, which is hsa-miR-508-3p. The sequence is represented by SEQ ID NO:1; and the miRNA molecular mark is abnormally high in expression in a placenta tissue of a pregnant woman suffering from the gestational diabetes and is secreted to the blood serum in an active secreting process of the tissue. The invention also provides a kit for diagnosing the gestational diabetes. The kit is characterized by comprising a reagent for detecting the level of the hsa-miR-508-3p in the blood serum; and the nucleotide sequence of the hsa-miR-508-3p is represented by SEQ ID NO:1.

Description

Technical field: [0001] The invention relates to a microRNA molecular marker for diagnosing gestational diabetes and a kit for diagnosing gestational diabetes. Background technique: [0002] In the second and third trimesters of pregnancy, with the increase of gestational age, the secretion of anti-insulin-like substances (such as: placental lactogen, estrogen, progesterone, etc.) in the pregnant woman's body is also increasing, which directly leads to the sensitivity of pregnant women to insulin. Decrease accordingly, in order to maintain the normal level of glucose metabolism, insulin secretion must increase accordingly, for those pregnant women with limited insulin secretion, due to the physiological change of not being able to metabolize normally, the blood sugar will eventually rise. We call this kind of clinical symptoms of hyperglycemia that occurred during pregnancy without diabetes before pregnancy as Gestational diabetes mellitus (GDM), which is a special type of d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/113C12N15/11
Inventor 周玲刘彦君张妍
Owner 周玲