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Survival motor neuron gene 1 (SMN1) detection primer set, kit and method capable of eliminating SMN2 interference

A gene detection and kit technology, applied in the field of molecular biology, can solve the problems of high cost, high dependence on equipment, and long time-consuming detection of gene mutation detection technology, so as to improve recognition ability, high sensitivity, reduce detection cost and popularize effect of difficulty

Pending Publication Date: 2020-05-22
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the above technical problems, the present invention provides a SMA gene detection primer set, kit and detection method that can eliminate SMN2 interference, and solve the problems of long time-consuming detection, high cost and high equipment dependence of existing gene mutation detection technology

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  • Survival motor neuron gene 1 (SMN1) detection primer set, kit and method capable of eliminating SMN2 interference
  • Survival motor neuron gene 1 (SMN1) detection primer set, kit and method capable of eliminating SMN2 interference
  • Survival motor neuron gene 1 (SMN1) detection primer set, kit and method capable of eliminating SMN2 interference

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Embodiment 1

[0050] The kit disclosed in this embodiment includes: Bst DNA Polymerase, dNTP, primer set, buffer, indicator and additive; wherein, Bst DNA Polymerase is 0.32U / μl, dNTP is 1.4mmol / L, and buffer consists of 20mmol / LTris -HCl, 10mmol / L (NH 4 ) 2 SO 4 , 50mmol / L KCl, 0.1% Tween-20, 8mmol / LMgSO in parts by mass 4 Composition, the indicator is 2×SYBR Green I, and the additive is composed of 0.1mol / L trehalose and 0.5mg / ml BSA. The primer set includes the following 7 primers:

[0051] F3: 5'-GCTATCTATGTCTATATAGCTAT-3';

[0052] B3: 5'-GTTTTGGCATCAAAATTCTTTAAT-3';

[0053] FIP-W: 5'-TGCTGGCAGACTTACTCCTTAACTTTATTTTTCCTTACAGGGTTVC-3';

[0054] FIP-M: 5'-TGCTGGCAGACTTACTCCTTAACTTTATTTTTCCTTACAGGGTTVT-3';

[0055] BIP-W: 5'-TGAAAGTGAATCTTACTTTTGTAAAAGTTTTACATTAACCTTTCAACTTVT-3';

[0056] LF: 5'-GTGAGCACCTTCCTTCTTTTTT-3';

[0057] LB: 5'-GTGGAAAACAAATGTTTTTGAAC-3'.

[0058] In the primer set of this embodiment, the concentrations of F3 and B3 are the same, both being 0.4 μmol / L...

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Abstract

The invention discloses a survival motor neuron gene 1 (SMN1) detection primer set, kit and method capable of eliminating SMN2 interference. Two different bases of an SMN1 and an SMN2 are respectivelydesigned at a 3' end of a forward inner primer (FIP) and a 3' end of a backward inner primer (BIP), and auxiliary mutation bases are respectively designed at the position of a second last base of the3' end of the FIP and the position of a second last base of the 3' end of the BIP, so that the specificity of loop-mediated isothermal amplification (LAMP) is enhanced. The influence of the SMN2 on amplification is eliminated by BIP-W, and FIP-W and FIP-M distinguish whether SMN1 mutates or not, and thereby, a genotype of the SMN1 of a to-be-detected sample is accurately judged. According to thedetection method disclosed by the invention, the loop-mediated isothermal amplification technology is employed, and has very high sensitivity, and accordingly, a buccal swab of a patient can be used as a template for amplification only by being boiled to release DNA without collecting a blood sample.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to an SMA gene detection primer set, a kit and a detection method capable of excluding SMN2 interference. Background technique [0002] Spinal muscular atrophy (SMA) is a relatively common autosomal recessive genetic disease characterized by muscle weakness and atrophy caused by degeneration and loss of motor neurons in the anterior horn of the spinal cord. The incidence rate of SMA is 1 / 10 000-1 / 6000, and the carrier rate is 1 / 50-1 / 40. The pathogenesis of SMA is mainly due to the mutation of the sixth base of exon 7 from C to T on the motor neuron survival gene 1 (survival motor neuron gene 1, SMN1) located on 5q13.2, resulting in functional defects of SMN protein hereditary neuromuscular disease. SMN is a widely expressed housekeeping protein that can bind to Sm protein as a subunit and recruit Sm nucleoproteins and small nuclear ribonucleic acids (snRNAs) ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6844C12N15/11
CPCC12Q1/6883C12Q1/6844C12Q2531/119Y02A50/30
Inventor 闫亚平张鑫
Owner SHAANXI NORMAL UNIV
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