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Dual-amplification method of template linear amplification and multi-biotin signal amplification

A linear amplification and double amplification technology, applied in the field of detection of vital signs substances, can solve the problems of non-linear amplification, false positive, and clinical inappropriateness, etc., and achieve the effect of low cost and reliable data

Inactive Publication Date: 2012-06-06
武汉中帜生物科技股份有限公司
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Problems solved by technology

However, due to some defects in the application process of PCR, such as nonlinear amplification, false positive and high cost of real-time PCR detection, it is not the most ideal way of application
[0004] The diagnosis of respiratory diseases can be achieved selectively through template amplification, but after template amplification, overnight hybridization detection is usually required, which is not a very suitable method for clinical practice

Method used

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Embodiment Construction

[0018] Further details will be given below in conjunction with the preferred embodiments shown in the accompanying drawings.

[0019] The double amplification method of template linear amplification and multi-biotin signal of the present invention is a double amplification method combining T7 amplification and multi-biotin signal amplification (MBSA). Wherein, the target nucleic acid is first linearly amplified by T7, and the amplified product is analyzed by MBSA. Different from PCR, T7 RNA linear amplification does not change the ratio of nucleic acid template, and the final product of the amplification is single-stranded RNA, which is then quantitatively detected by MBSA. MBSA is a highly sensitive detection method that can detect 100 target molecules by overnight hybridization. However, after the target is amplified by T7 RNA polymerase, the hybridization can be shortened to less than 2 hours.

[0020] The double amplification method includes the following steps:

[0021...

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Abstract

The invention relates to a dual-amplification method of template linear amplification and multi-biotin signal amplification. The method comprises steps that: nucleic acid is separated from a specimen; T7RNA polymerase linear amplification is carried out, such that an RNA product is obtained; the RNA product obtained through linear amplification is captured onto a microtiter plate, and signal amplification is carried out. The method is advantaged in low cost and more reliable data. With the method, the sensitivity and the detection time are similar to those of a PCR method.

Description

technical field [0001] The invention relates to the detection of vital substances, in particular to the detection of vital sign substances. Background technique [0002] On the one hand, respiratory diseases are caused by RNA (ribonucleic acid) viruses, DNA (deoxyribonucleic acid) viruses or bacteria. Determining the type of pathogen is critical to the selection of targeted drugs. Due to the small amount of pathogen nucleic acid in the specimen, template amplification such as PCR (polymerase chain reaction) or signal amplification such as bDNA technology (branched nucleic acid signal amplification technology) is required for detection. [0003] PCR is the method most commonly used for diagnosis. However, due to some defects in the application process of PCR, such as non-linear amplification, false positive and high cost of real-time PCR detection, it is not the most ideal way of application. [0004] The diagnosis of respiratory diseases can be achieved selectively throug...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李先强姜昕
Owner 武汉中帜生物科技股份有限公司
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