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Method and kit for evaluation and prediction of placenta-derived diseases

A placental source and kit technology, which is applied in biochemical equipment and methods, microbial determination/examination, medical automated diagnosis, etc., can solve the problems of lack of mature and reliable screening methods

Active Publication Date: 2021-08-13
安康优乐众成医疗器械有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In the prior art, a large number of clinical trials have been carried out to predict the risk of pregnancy complications related to placental dysplasia by using biochemical, biophysical detection and epidemiological statistics alone or in combination, but their sensitivity and specificity are also limited. It has not been possible to effectively assess and predict the risk of the aforementioned diseases at an earlier gestational age, and no mature and reliable screening methods have been applied clinically so far.

Method used

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  • Method and kit for evaluation and prediction of placenta-derived diseases
  • Method and kit for evaluation and prediction of placenta-derived diseases
  • Method and kit for evaluation and prediction of placenta-derived diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0249] Example 1: Using Qiagen kit to extract gDNA from placental tissue of pregnant women

[0250] Experimental Materials

[0251]Frozen placenta tissue (collected from Guangdong Provincial Maternal and Child Health Hospital, clinical research approval number: Yilun No. [201701044]);

[0252] Absolute ethanol (Beijing Chemical Industry, 00500);

[0253] Qiagen animal tissue extraction gDNA kit (Qiagen, 69504);

[0254] Qubit TM dsDNA HS Assay Kit (Thermo Fisher Scientific, Q32851);

[0255] λDNA HindIII digest (Takara, 3403)

[0256] experimental method

[0257] Placental tissue digestion: Take about 25 mg of placental tissue and prepare a digestion solution (180 μL buffer ALT + 20 μL proteinase K), mix it and incubate at 65°C for 4 hours until it is completely digested.

[0258] Cell lysis: vortex the digested product, add lysis buffer AL, absolute ethanol and RNaseA and incubate at room temperature for 10 minutes.

[0259] gDNA binding: Transfer all the samples af...

Embodiment 2

[0264] Example 2: Using MagMAX Cell-Free DNA Extraction Kit to Extract cfDNA from Maternal Plasma

[0265] Experimental Materials

[0266] Stored in Nanke Zhengtu MiniMax TM Peripheral blood from blood collection tubes such as cell-free DNA (cfDNA) noninvasive tubes (Apostle, Inc. Silicon Valley, California.), Cell-Free DNA BCT CE blood collection tubes (Streck, Inc., La Vista, NE) (collected from Maternal and Child Health Care of Guangdong Province Hospital, clinical research approval number: Yilun No. [201701044]);

[0267] Proteinase K (Qiagen, 19133);

[0268] SDS (Sigma, 74255-250G);

[0269] Dynabeads TM MyOne TM Silane (Thermo Fisher Scientific, 37002D);

[0270] MagMAX TM Cell Free DNA Lysis / Binding Solution (Thermo Fisher Scientific, A33600);

[0271] MagMAX TM Cell Free DNA Wash Solution (Thermo Fisher Scientific, A33601);

[0272] Absolute ethanol (Beijing Chemical Industry, 00500);

[0273] Qubit TM dsDNA HS Assay Kit (Thermo Fisher Scientific, Q3...

Embodiment 3

[0286] Example 3: Using the TequilaV3 DNA methylation method to build a library for gDNA or cfDNA

[0287] Experimental Materials

[0288] 7.5-10ng cfDNA or 200ng gDNA;

[0289] EZ DNA Methylation-Gold TM Kit(ZymoReserch, D5006);

[0290] Tequila V3 DNA Methylation Library Construction Kit (Ule Fusheng, EU-TQ-Methy);

[0291] Agencourt AMPure XP magnetic beads (Beckman, A63882);

[0292] EB buffer (Qiagen, 19086);

[0293] DEPC water (Solabao, R1600-500);

[0294] Absolute ethanol (Beijing Chemical Industry, 00500).

[0295] experimental method

[0296] Sulfite Conversion: According to EZ DNA Methylation-Gold TM The kit kit instruction manual prepares the CT conversion reagent and performs CT conversion treatment on the above-mentioned starting amount of cfDNA or gDNA. Wherein, the aforementioned cfDNA or gDNA is obtained by the method described in Example 2.

[0297] End repair: Use the Tequila V3 DNA Methylation Library Construction Kit to prepare end repair re...

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Abstract

The invention relates to a method and a kit for evaluation and prediction of placenta-derived diseases. Specifically, the technical scheme of the invention provides a method, a kit and a corresponding detection system for detecting the methylation modification level of at least one DNA methylation modification difference site or at least one DNA methylation haplotype on a given genome position in a sample to be detected, and the method can be used for accurately screening whether a subject suffers from the placenta-derived gestational disease or not in vitro.

Description

technical field [0001] The invention relates to the field of biological detection. Specifically, the present invention relates to risk assessment methods and kits for predicting pregnancy complications associated with placental dysplasia. More specifically, it relates to a method and a kit for assessing and predicting the risk of pregnant women suffering from gestational hypertension syndrome and other types of placental-derived diseases during the early pregnancy week (12-16 weeks of pregnancy). More specifically, the present invention relates to methods and kits for assessing and predicting the risk of preeclampsia. Background technique [0002] The formation of the placenta is a vital guarantee for normal pregnancy and healthy fetal growth in mammals. The placenta is the maternal-fetal interface, which plays an important role in providing necessary nutritional support for the fetus and regulating the immune tolerance of the mother. From the perspective of embryonic dev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G16H50/20
CPCC12Q1/6883G16H50/20C12Q2600/154G16H50/30G16B20/20
Inventor 张翼吴凯常璐媛刘佳金婉
Owner 安康优乐众成医疗器械有限责任公司
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