Method for qualitative and quantitative detection of microorganisms in human body
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A technology of microorganisms and target microorganisms, applied in the biological field, 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
Inactive Publication Date: 2016-06-15
JIANGHAN UNIVERSITY +1
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[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 human microorganisms
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Embodiment 1
[0039] Embodiment 1. Identification of human blood microorganisms
[0040] The samples to be tested are human tissues, body fluids and excreta, among which blood microorganisms are the basis for the diagnosis and treatment of human diseases. The sample to be tested in this embodiment is human blood, which is taken from a patient diagnosed with bacteremia by a doctor, and the detection of microorganisms in the blood is to provide a basis for a treatment plan.
[0041] Step 1. Determining the target microbial group in the sample to be tested, target microorganisms and non-target organisms, and reference microorganisms that do not exist in the sample to be tested, the specific method is as follows:
[0042] The number of target microbial groups ≥ 1, and each target microbial group includes ≥ 0 target microorganisms; target microorganisms can be bacteria, viruses, fungi, actinomycetes, rickettsia, mycoplasma, chlamydia, spirochetes and protozoa at least one of . The purpose of t...
Embodiment 2
[0089] Embodiment two, the identification of human feces microorganism
[0090] The sample to be tested in this embodiment is human feces, which are taken from patients with intestinal diseases diagnosed by doctors, and the detection of microorganisms in their feces is to provide a basis for treatment plans. This implementation is similar to the method in Example 1, and the methods, parameters and results not mentioned are the same as those in Example 1, so they will not be repeated here.
[0091] Step 1. Determining the target microbial groups, target microorganisms and non-target organisms in the test sample, and reference microorganisms that do not exist in the test sample.
[0092] The purpose of this embodiment is to identify Salmonella enterica in the sample to be tested. Its Latin formal name is Salmonella enterica. On NCBI (Nationalcenterforbiotechnologyinformation, National Center for Biotechnology Information), there are 33 physiological races of Salmonella enterica ...
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Abstract
The invention discloses a method for qualitative and quantitative detection of microorganisms in a human body and belongs to the technical field of the biology. The method comprises steps as follows: a target microorganism class group, a target microorganism and a non-target microorganism in a to-be-detected sample as well as reference microorganisms which don't exist in the to-be-detected sample are determined; characteristic regions of the target microorganism class group and the target microorganism are designed; multiple amplification primers of the characteristic regions are designed; the reference microorganisms and exogenous nucleic acid are added to the to-be-detected sample; and nucleic acid of the microorganisms in the to-be-detected sample is extracted; nucleic acid of the microorganisms is amplified through the designed multiple primers, and characteristic sequencing fragments are obtained through amplification; the microorganisms in the to-be-detected sample are analyzed qualitatively and quantitatively through the characteristic sequencing fragments. The microorganisms are not required to be pre-cultured and proliferated, multiple known microorganisms in the to-be-detected sample can be detected once in a high-throughput, high-accuracy and high-resolution manner, the detection process is simple and rapid, and the flow procedure is formal.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a qualitative and quantitative detection method for human microorganisms. Background technique [0002] Human microorganisms are an important basis for the diagnosis of human diseases, and accurate qualitative and quantitative detection of human microorganisms is very necessary. [0003] Existing qualitative and quantitative detection technologies for human microorganisms include morphological counting, chip detection, 16SrRNA sequencing, metagenomic sequencing and real-time quantitative PCR (Polymerase Chain Reaction, polymerase chain reaction). Morphological counting detection requires pre-cultivation of microorganisms, which takes a long time, non-culturable microorganisms cannot be detected, and only one type of microorganism can be detected at a time, the throughput is low, the sampling amount is limited when counting, and the results are rough, and it is impossible to classify ...
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