Serum lncRNA markers, primer sets, applications and kits for diagnosing unexplained recurrent miscarriage

A technology of recurrent miscarriage and primer set, applied in the field of genetic engineering, to achieve the effect of improving sensitivity and specificity, quantitative accuracy, fast and accurate disease state and disease severity

Active Publication Date: 2021-10-22
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no clinical report on the application of lncRNA in the auxiliary diagnosis of recurrent miscarriage. If the lncRNA related to recurrent miscarriage can be screened out as a biomarker for early diagnosis, and the corresponding diagnostic kits can be developed, it will be beneficial to the diagnosis of recurrent miscarriage. The present situation of diagnosis will definitely be a powerful impetus, and it will also open up a new way for its drug screening, drug efficacy evaluation and targeted therapy.

Method used

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  • Serum lncRNA markers, primer sets, applications and kits for diagnosing unexplained recurrent miscarriage
  • Serum lncRNA markers, primer sets, applications and kits for diagnosing unexplained recurrent miscarriage
  • Serum lncRNA markers, primer sets, applications and kits for diagnosing unexplained recurrent miscarriage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: lncRNA screening of decidua tissue

[0046] 1 material

[0047] 1.1 Selection of research objects and sample collection

[0048] (1) According to the diagnostic criteria: pregnant women with 2 or more consecutive pregnancy losses before 20 weeks of pregnancy or fetal weight less than 500g are the case group; according to age, gestational weeks, BMI and other indicators, the frequency matching method is used to select children Pregnant women with normal ability and non-medical reasons for miscarriage (without signs and signs of miscarriage, no history of miscarriage, and at least one normal child born) were used as the control group.

[0049] (2) B-ultrasound shows no germ or fetal heartbeat, and abortion caused by factors such as monogenic genetic diseases, polygenic genetic diseases, chromosomal abnormalities, and gene mutations must be excluded.

[0050] (3) Grouping of research objects:

[0051] Group A: the control group of pregnant women who chose abo...

Embodiment 2

[0061] Example 2: Detection of the expression levels of lnc-CHAC1-1, lnc-FMN1-1, lnc-TAX1BP1-4, lnc-C2CD4A-3, lnc-CES1-1, lnc-ATF3-3 in decidua tissue

[0062] 1 material

[0063] 1.1 Selection of research objects and sample collection

[0064] The selection of research objects and sample collection in this embodiment are the same as those in Embodiment 1.

[0065] 1.2 Reagents

[0066] The reagents used in qPCR are the same as in Example 1.

[0067] 2 methods

[0068] 2.1 Extraction of total RNA

[0069] The decidual tissues of 63 cases of normal control group and 63 cases of recurrent abortion cases were taken as objects.

[0070] (1) Take 50-100 mg of decidual tissue and grind it in liquid nitrogen, add 1 mL of TRIzol, homogenize it with a homogenate ultrasonic instrument, and place it at room temperature (15-30°C) for 5 minutes to completely separate the nucleoproteinase. (2) Add 200 μl chloroform for every 1 mL of TRIzol used, shake vigorously for 30 s, and place on...

Embodiment 3

[0085] Example 3: Detection of expression levels of lnc-CHAC1-1, lnc-FMN1-1, lnc-TAX1BP1-4, lnc-C2CD4A-3, lnc-CES1-1, lnc-ATF3-3 in serum

[0086] 1 material

[0087] 1.1 Selection of research objects and sample collection

[0088] The selection of research objects and sample collection in this embodiment are the same as those in Embodiment 1.

[0089] 1.2 Reagents

[0090] The reagents used in qPCR are the same as in Example 1.

[0091] 2 methods

[0092] 2.1 Extraction of total RNA

[0093] The serum samples of 63 cases of normal control group and 63 cases of recurrent miscarriage case group were taken as objects.

[0094] (1) Take 5ml of fresh heparin anticoagulant blood and centrifuge at 3000rpm for 5min in a centrifuge, take the supernatant and put it into clean 1.5ml EP tubes, 100μl per tube. (2) Add 900 μl TRIzol to the EP tube, shake and mix well, centrifuge at 12,000 rpm for 15 min at 4°C, and immediately transfer the supernatant to a clean 1.5ml EP tube. (3) A...

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Abstract

The invention belongs to the field of genetic engineering, and relates to a serum lncRNA marker, a primer set, an application and a kit for diagnosing unexplained recurrent miscarriage. The serum lncRNA markers include: the first lncRNA having more than 90% homology with the nucleotide sequence shown in SEQ ID NO: 1, and having more than 90% homology with the nucleotide sequence shown in SEQ ID NO: 2 The second lncRNA of sex, the third lncRNA having more than 90% homology with the nucleotide sequence shown in SEQ ID NO: 3, and the third lncRNA having more than 90% homology with the nucleotide sequence shown in SEQ ID NO: 4 The fourth lncRNA, the fifth lncRNA having more than 90% homology with the nucleotide sequence shown in SEQ ID NO: 5, and the sixth lncRNA having more than 90% homology with the nucleotide sequence shown in SEQ ID NO: 6 lncRNA. Serum lncRNA is a novel biomarker that can greatly improve the sensitivity and specificity of recurrent miscarriage diagnosis.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and more specifically relates to a serum lncRNA marker, a primer set, an application and a kit for diagnosing unexplained recurrent miscarriage. Background technique [0002] Recurrent spontaneous abortion (RSA) refers to clinically two or more consecutive pregnancy losses before 20W or fetal weight less than 500g, and its incidence rate accounts for about 1% to 3% of pregnancy outcomes. The etiology of RSA is extremely complex, and the etiology that has been identified so far includes chromosomal abnormalities, abnormal reproductive anatomical structures, endocrine disorders, reproductive system infections, autoimmunity, and environmental factors, but there are still 50% to 60% of RSA cases that cannot be identified clinically. It is called unexplained recurrent spontaneous abortion (URSA). Studies have shown that abnormal embryonic development is closely related to URSA. As a pathological p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 陆春城范赟黄振遥于明明杜桂珍傅广波夏彦恺王心如
Owner NANJING MEDICAL UNIV
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