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Microorganism detection method

A technology of microorganisms and pathogenic microorganisms, applied in the direction of microbial measurement/inspection, biochemical equipment and methods, resistance to vector-borne diseases, etc., can solve problems such as inability to obtain measurement results

Pending Publication Date: 2021-05-14
公益财团法人筑波医疗中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since biological samples contain various substances derived from living organisms in addition to nucleic acids, if nucleic acid amplification is performed without purification of biological samples, amplification inhibition or detection inhibition occurs, and in large In many cases, correct measurement results cannot be obtained

Method used

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preparation example Construction

[0120] [Method for preparing sample solution for nucleic acid amplification]

[0121] One embodiment of the present invention is a method for preparing a sample solution for nucleic acid amplification, the method comprising the following step (A):

[0122] (A) A step of preparing a mixed solution having an alkali concentration of 1 to 50 mM by mixing a biological sample or its suspension or solution with an alkaline solution.

[0123] In another embodiment, the method for preparing a sample solution for nucleic acid amplification includes the following step (B) in addition to the aforementioned step (A):

[0124] (B) A step of filtering the mixed solution prepared in the aforementioned step (A) and recovering the filtrate.

[0125] The preparation method is suitable for the preparation of a sample solution for nucleic acid amplification. The details of the steps (A), (B) and the like in this production method are the same as those of the steps (A), (B) and the like in the me...

Embodiment

[0131] The present invention will be specifically described below with reference to test examples, but the present invention is not limited to the test examples. The reagents used in the test examples are as follows.

[0132] KOD Mix: PCR reagent containing α-type polymerase

[0133] SCT Mix: Reagents containing primers and probes for Chlamydia trachomatis detection

[0134] MPN Mix: Reagents containing primers and probes for M. pneumoniae detection

[0135] IC Mix: Reagent containing internal control (IC)

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Abstract

The present invention addresses the problem of providing a method for detecting microorganisms in a biological sample without the need of nucleic acid purification which has disadvantages in that the process is complicated, takes a long time, uses an organic solvent, and needs expensive reagents. This method for detecting microorganisms in a biological sample comprises the steps (A) and (C): (A) preparing a liquid mixture having an alkali concentration of 1 to 50 mM by mixing a biological sample or a suspension or solution thereof with an alkaline solution; and (C) amplifying a nucleic acid of a microorganism in the liquid mixture prepared in the step (A).

Description

technical field [0001] The present invention relates to a method for detecting microorganisms contained in a biological sample. Background technique [0002] In clinical diagnosis, etc., it is important to detect microorganisms contained in biological samples, and the following method is performed: using PCR or the like to amplify the nucleic acid of the microorganisms to be detected in the biological sample, and detecting the amplified nucleic acid to identify the presence or absence of microorganism. However, since biological samples contain various substances derived from living organisms in addition to nucleic acids, if nucleic acid amplification is performed without purification of biological samples, amplification inhibition or detection inhibition occurs, and in large In many cases, correct measurement results cannot be obtained. Therefore, when detecting microorganisms contained in a biological sample, the biological sample is usually pretreated instead of being di...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/6844
CPCC12Q1/04C12Q1/6844Y02A50/30
Inventor 上仓佳子杉本道俊杉本泰康铃木广道
Owner 公益财团法人筑波医疗中心