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Compositions, kits, methods and systems for nucleic acid sample amplification

A nucleic acid sample and composition technology, applied in the field of nucleic acid sample amplification composition, can solve the problems of difficulty in implementing multiple PCR capture technology, primer coverage, uniformity, etc.

Pending Publication Date: 2022-07-01
GUANGDONG FAPON BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the more target regions to be captured by multiplex PCR, the coverage and uniformity of primer amplification will be affected, and the more difficult it is to realize the multiplex PCR capture technology

Method used

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  • Compositions, kits, methods and systems for nucleic acid sample amplification
  • Compositions, kits, methods and systems for nucleic acid sample amplification
  • Compositions, kits, methods and systems for nucleic acid sample amplification

Examples

Experimental program
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Effect test

Embodiment 1

[0064] Compositions for amplification of nucleic acid samples, including primer set 9. Among them, phenylketonuria (Phenylketonuria) is a disease caused by PAH gene defect. Primers are creatively designed for 212 pathogenic loci on PAH to avoid primer dimers and specific amplification, taking into account high homogeneity and High coverage, and optimized to obtain primer set 9 (nucleotide sequences shown in SEQ ID NO: 163-SEQ ID NO: 190).

Embodiment 2

[0066] A composition for nucleic acid sample amplification, including primer set 9 and primer set 2. Among them, 21-hydroxylase deficient congenital adrenal hyperplasia (Adrenal hyperplasia, congenital, due to 21-hydroxylasedeficiency) is a disease caused by CYP21A2 gene defect, and primers are creatively designed for 10 pathogenic loci on CYP21A2 , avoiding primer-dimer and specific amplification, taking into account high uniformity and high coverage, and optimized to obtain primer set 2 (nucleotide sequence shown in SEQ ID NO:47-SEQ ID NO:60); primers Group 9 is the same as Example 1.

Embodiment 3

[0068] A composition for nucleic acid sample amplification, comprising primer set 9 and primer set 5. Among them, Glycogen Storage Disease II (Glycogen Storage Disease II) is a disease caused by GAA gene defect. Primers are creatively designed for 75 pathogenic loci on GAA to avoid primer dimerization and specific amplification. Taking into account high homogeneity and high coverage, primer set 5 (nucleotide sequences shown in SEQ ID NO: 97-SEQ ID NO: 138) was obtained by optimization; primer set 9 was the same as Example 1.

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Abstract

The invention relates to a composition, a kit, a nucleic acid amplification method, a library building method and a system for nucleic acid sample amplification, which can be used for detection of single gene disease related variation sites, and high homogeneity and / or high coverage can be obtained while primer dimer and non-specific amplification are avoided.

Description

technical field [0001] The present invention relates to the field of molecular detection, in particular, to compositions, kits, methods and systems for nucleic acid sample amplification. Background technique [0002] Monogenic genetic disease (abbreviation: monogenic disease) is a disease caused by a single gene defect, in line with Mendelian inheritance, so it is also called Mendelian genetic disease. It is a type of disease that causes serious damage to human health (death, disability or teratogenicity) and lacks effective diagnosis and treatment methods, and is also one of the main causes of birth defects. So far, more than 10,000 kinds of monogenic diseases have been discovered. According to the statistics of the World Health Organization (WHO), the comprehensive incidence rate is as high as 1 / 100. Monogenic diseases often do not show structural deformities during fetal development, and are difficult to detect in routine obstetric examinations, and many recessive geneti...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 李洪洲马晓冰章瑞程关媛妹张钰王晓丽
Owner GUANGDONG FAPON BIOTECH CO LTD
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