Primer combination for detecting microdeletion or/and microduplication of recurrent abortion, MLPA probe, gene chip and kit

A technique of recurrent miscarriage and primer combination, applied in the field of MLPA probes, can solve the problem of low detection rate of recurrent miscarriage, and achieve the effects of enriching the causative gene map, low cost and good specificity

Active Publication Date: 2020-03-03
国家卫生健康委科学技术研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of low detection rate of recurrent miscarriage caused by micro-deletion / and micro-duplication in the prior art and to determine whether the biological sample to be tested is a recurrent miscarriage biological sample The detection of recurrent miscarriage is more complete and accurate. The present invention provides a MLPA probe combination, kit, gene chip and system that can quickly screen and detect whether the biological sample is susceptible to recurrent miscarriage. The kit, MLPA probe provided by the present invention Needles, gene chips and systems have low requirements for biological samples to be tested, simple methods, low cost, and high clinical application value

Method used

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  • Primer combination for detecting microdeletion or/and microduplication of recurrent abortion, MLPA probe, gene chip and kit
  • Primer combination for detecting microdeletion or/and microduplication of recurrent abortion, MLPA probe, gene chip and kit
  • Primer combination for detecting microdeletion or/and microduplication of recurrent abortion, MLPA probe, gene chip and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1 Primer, probe and test kit

[0058] 1. Primer combination

[0059] According to the sequence information of the internal reference gene GAPDH and the target genes AKT1 and PDZD2, primers were designed, and the designed primers were evaluated for secondary structure and Tm value, and finally obtained good specificity, high sensitivity, and detection under the same reaction conditions. The primer sequences of the nucleotide sequences shown in SEQ ID NO.1-21 of the present invention for the recurrent abortion microdeletion or / and microduplication of the biological sample to be tested are shown in Table 1 in turn:

[0060] Table 1 primer combination sequence provided by the present invention

[0061]

[0062] Secondary structure evaluation and Tm value evaluation can be carried out by any method commonly used in the art. For example, in one embodiment of the present invention, DNA folding form is used to evaluate its secondary structure, specifically (http:...

Embodiment 2

[0077] Example 2 System for rapid screening of biological samples susceptible to recurrent miscarriage

[0078] Such as figure 2 As shown, the system for the rapid screening of biological samples prone to recurrent miscarriage provided by the invention comprises:

[0079] 101: whole genome DNA extraction unit, used to extract the whole genome DNA sample in the biological sample; 102: whole genome DNA sample processing unit, used to denature the whole genome DNA sample in the whole genome DNA extraction unit, and combine with the whole genome DNA sample The MLPA probe combination made by the primer combination or the MLPA probe combination or the MLPA probe combination in the kit is hybridized, and finally the hybridization product is amplified to obtain an analysis sample; 103: sample analysis unit, used for judgment analysis Whether the sample is a biological sample susceptible to recurrent miscarriage, and give the judgment result.

[0080] Specifically, the 102: whole-ge...

Embodiment 3

[0082] Example 3 Screening and detection of biological samples with normal DNA

[0083] The kit in Example 1 is used to detect blood biological samples with normal DNA. In order to reflect the high sensitivity and high specificity of the MLPA probe combination provided by the present invention, the application also designed non-target genes TBX6, LHX1, HNF1B, HFE2 , GSTT1, CRKT1, CRKL, DUSPP22 and WNT3 MLPA probes, the MLPA probe sequences of the above genes are shown in Table 1, and added to the above MLPA probe combinations, the specific operations of screening and detection are as follows:

[0084] Table 2 MLPA probes for other genes

[0085]

[0086]

[0087]

[0088]

[0089] 1. Sample acquisition and processing

[0090] Peripheral blood was collected by conventional methods. The peripheral blood is an isolated biological sample taken from the human body, and the whole genome DNA in the sample was extracted using a conventional DNA extraction kit to obtain a ...

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Abstract

The invention provides a primer combination for detecting microdeletion or / and microduplication of recurrent abortion, an MLPA probe, a gene chip and a kit, and relates to the field of biotechnology.The primer combination has a nucleotide sequence as shown in SEQ ID NO.1-21, can be made into MLPA probes, gene chips, kits or constructs, and recombinant cells, and can also be used as a main component of a system for rapidly screening biological samples susceptible to recurrent abortion. The primer combination provided by the invention can help simultaneously detect recurrent abortion microdeletion or / and microduplication at multiple sites, has good specificity, sensitive response, short detection time and simple method, can detect from point mutation to large chromosome deletion / insertion,has low cost and high clinical application value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer combination for detecting biological samples susceptible to recurrent miscarriage and the MLPA probe, gene chip, kit, system and method prepared therefrom. Background technique [0002] Recurrent Spontaneous Abortion (RSA) is a common pathological pregnancy in gynecology, which refers to 2 or more consecutive spontaneous abortions. Studies have shown that there are many reasons for recurrent miscarriage, among which genetic factors are closely related to the occurrence of recurrent miscarriage, and chromosomal microdeletions and microduplications are also common types of genetic factors. Chromosomal microdeletion / microduplication refers to the deletion or duplication of 1.5kb-10Mb in length on the chromosome. Although the incidence of each microdeletion or / and microduplication syndrome is very low, due to the limitation of clinical detection technology, a large number of pa...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6844C12N15/11C12N5/10
CPCC12Q1/6883C12Q1/6844C12Q2600/16C12Q2537/143C12Q2561/125
Inventor 王彬彬王晶
Owner 国家卫生健康委科学技术研究所
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