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Primer combination for detecting infectious diarrhea pathogen and kit thereof

A primer composition and a technology for infectious diarrhea, applied in the field of nucleic acid amplification, can solve the problems of high false negative rate and false positive rate, difficulty in detecting difficult-to-cultivate pathogens, and long detection time (generally about 2-3 days).

Active Publication Date: 2017-07-04
SHANGHAI IGENETEC DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the analysis of routine clinical biochemical indicators cannot accurately identify the pathogens infected by patients, most clinical treatments are still in the stage of empirical medication, and there is a large blindness in the use of advanced antibiotics
The currently popular method of identifying pathogens in clinical practice mainly relies on the bacterial culture detection method, but the bacterial culture detection method has relatively large defects: long detection time (generally about 2-3 days), low accuracy rate (false negative rate and false negative rate) The positive rate is high), it is difficult to detect pathogens that are difficult to cultivate, etc.

Method used

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  • Primer combination for detecting infectious diarrhea pathogen and kit thereof
  • Primer combination for detecting infectious diarrhea pathogen and kit thereof
  • Primer combination for detecting infectious diarrhea pathogen and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] This example describes the microfluidic chip used in the present invention.

[0057] Such as figure 1 As shown, the microfluidic chip used in the present invention is a disc-shaped microfluidic chip, which includes 4 reaction detection areas 11, and each reaction detection area 11 includes a sequentially connected sample pool 12, distribution pool 13, capillary micro The valve 15 and the amplification pool 16, the sampling pool 12 communicates with the distribution pool 13 through an arc-shaped channel, the sampling pool is provided with a sampling hole, and the distribution pool 13 communicates with the waste liquid pool and the exhaust hole through an arc-shaped channel ; Each reaction detection area 11 is equipped with 8 amplification pools 16 .

[0058] Among them, the main function of the sampling pool 12 is to load the reaction solution; the main function of the distribution pool 13 is to evenly distribute the reaction solution to the amplification pool; the ampl...

Embodiment 2

[0061] This example is the primer composition and kit used in the present invention for detecting infectious diarrhea pathogens.

[0062] The primer composition of the present invention includes at least one of six primer sets packaged independently. The infectious diarrhea pathogen includes at least one of the following six pathogens: rotavirus, enteric adenovirus, norovirus, salmonella, shigella, and campylobacter jejuni. The six primer sets corresponding to the above-mentioned pathogens are rotavirus primer set, intestinal adenovirus primer set, norovirus primer set, salmonella primer set, shigella primer set and campylobacter jejuni primer set. Included is an internal control primer set for amplifying the human hemoglobin-beta chain. See Table 1 for details of the primer sequences of the above primer sets.

[0063] The present invention relates to a kit containing the above primer composition, which also includes the microfluidic chip coated with the above primer set as ...

Embodiment 3

[0066] This embodiment is a method for detecting infectious diarrhea pathogens using the primer composition and kit described in Embodiment 2, which includes the following steps:

[0067] 1. Coating of primer composition;

[0068] Such as figure 2 and image 3 As indicated, the rotavirus primer set, intestinal adenovirus primer set, norovirus primer set, salmonella primer set, shigella primer set, campylobacter jejuni primer set, and internal control primer set were mixed with sucrose respectively to prepare into a corresponding mixed solution, and the final concentrations of each primer group and sucrose in the mixed solution were 0.15 μM and 1.0% (mass percentage); get 1 μ L of the mixed solution and put it into the corresponding amplification pool of the microfluidic chip (amplification pool 1, 2, 3, 4, 5, 6, 8), the amplification pool 7 stores RNase-free water, and the microfluidic chip is dried in a 37°C oven, pressed and sealed, and after stamping and vacuuming, the p...

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Abstract

The invention relates to a primer combination for detecting infectious diarrhea pathogen. The primer combination comprises at least one combination of the following combinations: a rotavirus primer combination, an enteric adenovirus primer combination, a Norovirus primer combination, a salmonella primer combination, a Shigella primer combination and a campylobacter jejuni primer combination. The invention also relates to a kit containing the primer combination. The kit also comprises a primer-coating micro-fluidic chip, an isothermal amplification reaction liquid, an isothermal amplification enzyme solution, and a negative control. The invention also relates to a detection method by using the above primer combination. The detection method comprises steps as follows: coating of the primer combination, nucleic acid extraction of a sample to be detected, LAMP reaction and result interpretation. The kit provided by the invention can detect infectious diarrhea pathogen rapidly and accurately within one hour, and is also of great significance for rapid assisted guide treatment and drug use. The multi-index detection also can be used in regional epidemiological investigation and epidemic surveillance so as to study epidemic situation of infectious diarrhea in China.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid amplification, and in particular relates to a primer composition for detecting infectious diarrhea pathogens, a microfluidic chip system-based kit for detecting infectious diarrhea pathogens and a method thereof. Background technique [0002] Infectious diarrhea is the most common type of disease in the world. Every year, an estimated 3.2 million children aged ≤5 die from diarrhea, accounting for 24.8% of the deaths of children of the same age. In developing countries, each child suffers from 1-12 illnesses per year In developed countries, every child gets sick 1-5 times a year. This shows that infectious diarrhea is still an important disease at present. Infectious diarrhea refers to diarrhea caused by intestinal inflammation caused by various acute and chronic bacterial, viral, fungal, and parasitic infections. We also refer to infectious diarrhea other than cholera, bacillary and amoebic ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/10C12Q1/04C12N15/11
CPCC12Q1/6844C12Q1/689C12Q1/701C12Q2600/166C12Q2531/119C12Q2545/101C12Q2565/629Y02A50/30
Inventor 方雪恩李新鑫郭如威李平丁钦孔继烈
Owner SHANGHAI IGENETEC DIAGNOSTICS CO LTD
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