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Method and device for detecting microorganisms in blood

A microbial and blood technology, applied in the field of microbial detection, can solve the problems of cumbersome and time-consuming culture techniques, and achieve the effect of relieving pain, alleviating pain, and benefiting healthy blood transfusion

Pending Publication Date: 2016-04-27
深圳华大因源医药科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These techniques have played a huge role in clinical diagnosis, but there are still some shortcomings, such as cumbersome and time-consuming culture techniques, and histological biopsy and surgical exploration may bring pain and burden to patients. Apply new technology to compensate

Method used

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  • Method and device for detecting microorganisms in blood
  • Method and device for detecting microorganisms in blood
  • Method and device for detecting microorganisms in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Determination of the free nucleic acid sequence of the plasma sample

[0036] The blood / plasma samples of patients with fever of unknown origin came from the Inner Mongolia Maternal and Child Health Hospital. After the nucleic acid was extracted, the library construction and sequencing were carried out according to the library construction and sequencing instructions of BGI’s microchip semiconductor sequencing platform BGISEQ100.

[0037] This detection technology uses a new generation of high-throughput sequencing technology to detect pathogens in the plasma of patients with fever of unknown origin. The specific implementation steps are as follows:

[0038] Extraction of nucleic acid from plasma samples

[0039] Use Tiangen's Micro Sample Genome Extraction Kit to extract nucleic acid from plasma samples according to the instructions in the kit.

[0040] 2. DNA library construction

[0041] 2.1 End Repair

[0042] 2.1.1 Take out 10×PolynucleotideKinase (PNK...

Embodiment 2

[0064] Example 2 Microorganism / Pathogen Detection

[0065] The overall detection and analysis process is as follows: image 3 shown.

[0066] According to steps 1-4 of the method in Embodiment 1, the sequence determination data of the plasma sample nucleic acid is obtained, and the sequence data consists of multiple reads of different lengths.

[0067] 5. Data preprocessing

[0068] 5.1 Filter raw data, for BGISEQ100 sequencing data, remove reads less than 50bp.

[0069] 5.2 Filter host human genetic material. Most of the plasma samples are cell-free DNA of the host, the data shows that it is as high as more than 95%, and the content of the remaining microorganisms is less than 1%. In order to avoid the influence of the host’s genetic material when detecting pathogenic microorganisms, TMAP software and human reference genome reference sequences are used Perform comparison, mask out the sequence on the comparison, and obtain non-human sequence data.

[0070] 6. Pathogen da...

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PUM

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Abstract

The invention provides a method for detecting microorganisms in plasma as well as a method and a device for detecting host infection pathogens. The method for detecting the microorganisms in the plasma includes extracting host plasma sample nucleic acid and subjecting at least part of the extracted nucleic acid to sequencing so as to obtain sequencing data; removing host sequencing data from the sequencing data; comparing the rest of sequencing data to a microorganism reference database so as to obtain comparison results, wherein the microorganism reference database includes reference sequences of multiple kinds of bacteria and multiple kinds of viruses; determining microorganism species in the plasma and richness of the microorganism species according to the comparison results. By the methods or the device, the microorganisms in the plasma can be detected rapidly. The methods and the device can be used for assisting clinical diagnosis.

Description

technical field [0001] The invention relates to microorganism detection, in particular to a method for detecting microorganisms in blood, a method for detecting host infection pathogens and a device thereof. Background technique [0002] Based on the relationship between the host and the microorganisms, the microorganisms in the host can be divided into many types, including normal flora, symbiotic microorganisms including parasitic microorganisms with the host, microorganisms that cause infection and pathogenicity of the body-pathogens, etc. The plasma of the host contains cell-free DNA, including cell-free DNA fragments derived from the host and derived from microorganisms. [0003] Fever of unknown origin (fever of unknown origin, fever of unknown origin, FUO) refers to a disease that has recurrent fever exceeding 38.3°C and lasts for more than 3 weeks, and cannot be clearly diagnosed after 1 week of detailed examination. According to the potential etiology, FUO can be d...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04C12M1/00
Inventor 栗东芳宫艳萍张印新路惠萍韩秋莲
Owner 深圳华大因源医药科技有限公司
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