Detection kit for virulence gene of spinal muscular atrophy and application thereof

A spinal muscular atrophy, detection kit technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of indistinguishable, expensive, time-consuming, etc., and achieves simple operation, low detection cost, Detecting fast effects

Inactive Publication Date: 2017-01-11
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PCR-RFLP, allele-specific amplification, and HRM techniques are only suitable for detecting homozygous deletions of the SMN1 gene, but cannot distinguish SMA pathogenic gene carriers with heterozygous deletions from normal individuals without deletions
MLPA technology is currently the mainstream technology for SMA laboratory diagnosis. This method uses the SMA MLPA detection kit from MRC Company in the Netherlands, which is expensive (>150 yuan / sample), and requires expensive analytical instruments such as capillary electrophoresis, which takes a long time and is not suitable for For screening of large populations

Method used

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  • Detection kit for virulence gene of spinal muscular atrophy and application thereof
  • Detection kit for virulence gene of spinal muscular atrophy and application thereof
  • Detection kit for virulence gene of spinal muscular atrophy and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Spinal Muscular Atrophy Gene Detection Kit

[0047] The spinal muscular atrophy pathogenic gene detection kit of the present invention comprises:

[0048] Primer SMN-Ex7-F, its sequence is:

[0049] 5'-CCGCTATCTATATATAGCTATCTATG-3' (SEQ ID NO.1);

[0050] Primer SMN-Ex7-R, its sequence is:

[0051] 5'-CCTACATTAACCTTTCAACTTTT-3' (SEQ ID NO.2);

[0052] Probe SMN-Ex7-P, its sequence is:

[0053] 5'-FAM-CTTACAGGGTTTCAGACAAAAT-MGB-3' (SEQ ID NO. 3).

[0054] Further, the spinal muscular atrophy pathogenic gene detection kit of the present invention also includes:

[0055] Primer SMN-In7-F, its sequence is:

[0056] 5'-CCTATTTTCCTTACAGGGTTTC-3' (SEQ ID NO.4);

[0057] Primer SMN-In7-R, its sequence is:

[0058] 5'-TTGTGAAAGTATGTTTCTTCCACAT-3' (SEQ ID NO.5);

[0059] Probe SMN-In7-P, its sequence is:

[0060] 5'-FAM-CCTTTCAACTTTTTTAACATCT-MGB-3' (SEQ ID NO. 6).

[0061] Further, the spinal muscular atrophy pathogenic gene detection kit of the present inve...

Embodiment 2

[0071] Example 2 Application of Spinal Muscular Atrophy Gene Detection Kit

[0072] 1. Obtain the genomic DNA of the person to be tested

[0073] From the genomic DNA preserved in the sample resource bank of the genetic center, select 1050 samples (including 50 cases of homozygous deletion SMA patients, 500 cases of heterozygous deletion SMA carriers and 500 cases) that have been tested by MLPA method. normal people).

[0074] Blind analysis: use NanoDrop ND-2000 to measure the concentration of the above-mentioned genomic DNA, take the concentration at 20-150ng / μL, A 260 / A 280 Samples between 1.8 and 1.9 will be blindly numbered as the genomic DNA of the person to be tested for future use; for samples that do not meet the above requirements, they will not be included in this analysis and test.

[0075] 2. Accurate quantification of the genomic DNA of the subject to be tested

[0076] 1. Preparation of working solution (Qubit working solution)

[0077]The DNA-binding dye ...

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Abstract

The invention provides a detection kit for virulence gene of spinal muscular atrophy and application thereof, comprising primer SMN-Ex7-F with such sequences as SEQ ID NO.1; primer SMN-Ex7-R with such sequences as SEQ ID NO.2 and probe SMN-Ex7-P with such sequences as SEQ ID NO.3. The above kit in the invention can be applied to detect the carrier of spinal muscular atrophy virulence gene, comprising the steps of extracting the genome DNA from subject to be detected; precisely quantifying the genome DNA from subject to be detected; and performing primary fluorescent quantitation PCR based on genome DNA from subject to be detected as the template to obtain the primary fluorescent quantitation curve. The kit in the invention can visually, precisely, quickly and succinctly detect the SMN1 gene, so as to realize the distinguishing of SMA virulence gene carrier and normal people.

Description

technical field [0001] The invention relates to a detection kit, in particular to a spinal muscular atrophy pathogenic gene detection kit and application thereof. Background technique [0002] Spinal muscular atrophy (SMA) is an autosomal recessive genetic disease. It is a neuromuscular genetic disease caused by genetic mutations that lead to degeneration of the anterior horn cells of the spinal cord. The clinical manifestations are progressive, symmetrical proximal limb and trunk muscle weakness. , atrophied, and the patient eventually died of respiratory failure and severe lung infection. SMA is the second most fatal autosomal recessive genetic disease with no effective treatment. The incidence of SMA in live births is about 1 / 6000-1 / 10000, and the carrier rate in the population is 1 / 35-1 / 80. In view of the high carrier rate and morbidity rate of SMA in the Chinese population, marriage and fertility guidance through SMA carrier screening is the most effective way to bloc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q2531/113C12Q2537/16C12Q2561/101C12Q2545/101
Inventor 梅世月白楠赵振华孔祥东
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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