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A qualitative and quantitative detection method for non-diagnostic microorganisms

A technology for target microorganisms and microorganisms, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of inaccurate qualitative and quantitative detection of microorganisms, and achieve the effect of fine detection results, simple detection process, and probability guarantee

Active Publication Date: 2019-11-15
JIANGHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem of inaccurate qualitative and quantitative detection of microorganisms in the prior art, the embodiment of the present invention provides a qualitative and quantitative detection method for non-diagnostic microorganisms

Method used

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  • A qualitative and quantitative detection method for non-diagnostic microorganisms
  • A qualitative and quantitative detection method for non-diagnostic microorganisms
  • A qualitative and quantitative detection method for non-diagnostic microorganisms

Examples

Experimental program
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Embodiment 1

[0039] Embodiment 1. Identification of rice bacterial blight

[0040] Example 1 of the present invention From the Lingshui Experimental Base of Hainan, Chinese Academy of Sciences, rice leaves naturally infected with bacterial blight were taken as samples to be tested. Bacteria were qualitatively and quantitatively detected.

[0041] Step 1: Determine the target microbial groups, target microorganisms and non-target organisms in the sample to be tested, and reference microorganisms that do not exist in the sample to be tested. The specific methods are as follows:

[0042] The number of target microbial groups is ≥ 1, and each target microbial group includes ≥ 0 target microorganisms; target microorganisms can be bacteria, viruses, fungi, actinomycetes, rickettsia, mycoplasma, chlamydia, spirochetes, algae and protists at least one of animals. The purpose of the embodiment of the present invention is to identify rice bacterial blight, therefore, only concerned about the bacte...

Embodiment 2

[0088] Embodiment 2. Identification of Human Fecal Microorganisms

[0089] The sample to be tested in the second embodiment of the present invention is human feces, which is taken from a patient diagnosed by a doctor with intestinal diseases, and the detection of microorganisms in the feces is to provide a basis for a treatment plan. This implementation is similar to the method in the first embodiment, and the methods, parameters and results that are not mentioned are the same as those in the first embodiment, so they will not be repeated.

[0090] Step 1: Determine target microbial groups, target microorganisms and non-target organisms in the sample to be tested, and reference microorganisms that do not exist in the sample to be tested.

[0091] The purpose of this example is to identify Salmonella enterica in the sample to be tested, its Latin name is Salmonellaenterica, and on NCBI (National center for biotechnology information, National Center for Biotechnology Information...

Embodiment 3

[0106] Example 3. Identification of microorganisms in triangular lake water

[0107] The sample to be tested in this embodiment is the water body in the Triangle Lake in Wuhan, Hubei Province. The purpose of detecting microorganisms in the water body is to monitor the pollution of the water body by human and animal feces through the amount of microorganisms. This implementation is similar to the method in the first embodiment, and the methods, parameters and results that are not mentioned are the same as those in the first embodiment, so they will not be repeated.

[0108] Step 1: Determine the target microbial groups, target microorganisms and non-target organisms in the sample to be tested, and reference microorganisms that do not exist in the sample to be tested. The specific methods and results are the same as those in the second embodiment.

[0109] Step 2: According to the reference genome sequence of the target microorganism group, the reference genome sequence of the t...

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Abstract

The invention discloses a qualitative and quantitative detection method for non-diagnostic microorganisms, which belongs to the field of biotechnology. The method comprises the following steps: determining target microbial groups, target microorganisms and non-target organisms, and reference microorganisms that do not exist in the test sample; designing target microbial groups and target microbial characteristic regions; designing Multiple amplification primers for the characteristic region; after adding reference microorganisms and exogenous nucleic acids to the sample to be tested, extract the nucleic acid of the microorganisms in the sample to be tested; use the designed multiple primers to amplify the microbial nucleic acid, and obtain a characteristic sequencing fragment; Characteristic sequencing fragments qualitatively and quantitatively analyze the microorganisms in the sample to be tested. The present invention does not need to pre-cultivate and proliferate microorganisms, and can perform high-throughput, high-accuracy, and high-resolution detection of multiple known microorganisms in a sample to be tested at one time, and the detection process is simple, fast and standardized.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a qualitative and quantitative detection method of microorganisms for non-diagnostic purposes. Background technique [0002] Microorganisms are ubiquitous, and their detection applications are also very extensive, including agricultural disease identification and prediction, food safety testing, and medical pathogen detection. [0003] Existing microbial qualitative and quantitative detection technologies include morphological counting, chip detection, 16S rRNA sequencing, metagenomic sequencing and real-time quantitative PCR (Polymerase Chain Reaction, polymerase chain reaction). Morphological counting detection requires pre-cultivation of microorganisms, which is time-consuming, unculturable microorganisms cannot be detected, only one microorganism can be detected at a time, the throughput is low, the sampling amount during counting is limited, and the results are rough, which cann...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869C12Q1/06C12Q1/04C12Q1/70
CPCC12Q1/6869C12Q2531/113C12Q2537/143C12Q2535/122
Inventor 彭海
Owner JIANGHAN UNIVERSITY
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