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Molecular marker used for diagnosing extremely severe case of hand-foot-and-mouth disease and testing method as well as kit

A molecular marker, hand, foot and mouth disease technology, applied in the field of biological detection technology and medical diagnosis, can solve the problems of distinguishing hand, foot and mouth disease, poor prognosis of children, etc.

Active Publication Date: 2014-10-01
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, because HFMD is highly contagious to infants and young children, HFMD is generally diagnosed through clinical symptoms and virus identification, but there is no clear molecular marker to distinguish mild and severe HFMD, causing many children poor prognosis

Method used

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  • Molecular marker used for diagnosing extremely severe case of hand-foot-and-mouth disease and testing method as well as kit
  • Molecular marker used for diagnosing extremely severe case of hand-foot-and-mouth disease and testing method as well as kit
  • Molecular marker used for diagnosing extremely severe case of hand-foot-and-mouth disease and testing method as well as kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The method for rapidly and accurately diagnosing children with extremely severe hand, foot and mouth disease comprises the following steps:

[0043] 1. Collect blood from 5 healthy children, 5 children with mild HFMD and 5 children with severe HFMD, centrifuge at 1000×g for 10 minutes, and then take out the serum;

[0044] 2. According to the instructions of the exosome extraction kit, extract the exosome in the serum;

[0045] 3. Observe the shape of the exosome under the transmission electron microscope, which is nearly round, with a diameter between 30-100nm. In addition, the expression of two biomarker proteins CD63 and TSG101 of exosome was detected by western Blot. The results show that CD63 and TSG101 in exosome are significantly enriched than those in serum. The above results show that we have successfully extracted exosome, see figure 1 .

[0046] 4. Extract the RNA in the exosome with Trizol reagent, and measure its concentration and purity;

[0047] 5. Pe...

Embodiment 2

[0069] In order to further confirm whether these differential miRNAs (miR-671-5p, miR-16-5p and miR-150-3p) are closely related to the rapid and accurate diagnosis of severe HFMD, 18 healthy children, 18 The RNA of exosome in the serum of children with mild HFMD and 18 children with extremely severe HFMD, the RNA of exosome in the serum of these groups was verified by the fluorescence quantitative PCR evaluation method described in Example 1, and the serum of healthy children Exosome RNA was used as a control, and the results were as follows image 3 and Figure 4 As shown, the experimental results showed that miR-671-5p, miR-16-5p and miR-150-3p in children with extremely severe HFMD were significantly changed compared with healthy children and children with mild HFMD. In particular, miR-671-5p is expressed in the serum exosome of normal children and children with mild HFMD, but is almost not expressed in the serum exosome of children with extremely severe HFMD. miR-671-5p c...

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Abstract

The invention discloses a molecular marker used for diagnosing the extremely severe case of a hand-foot-and-mouth disease. The molecular marker comprises miR-671-5p, miR-16-5p and miR-150-3p. A biomarker related by the invention and used for diagnosing the extremely severe case of the hand-foot-and-mouth disease has the advantages of specificity distinguishing capacity, good stability, and high efficiency; besides, the biomarker has a better property of diagnosing indexes; moreover, the testing kit which is related by the invention and used for diagnosing the extremely severe case is low in cost. Therefore, the kit can be popularized on a large-scale.

Description

technical field [0001] The invention relates to the fields of biological detection technology and medical diagnosis, in particular to a molecular marker for diagnosing extremely severe hand, foot and mouth disease, a detection method and a kit. Background technique [0002] Hand, foot, and mouth disease (HFMD) is a common infectious disease, mainly affecting infants and young children. The pathogens causing hand, foot and mouth disease are mainly Coxsackie virus (Coxasckie virus) A group 16, 4, 5, 7, 9, 10 types of Enterovirus, B group 2, 5, 13 types; viruses) and enterovirus 71 (EV71), of which EV71 and CA16 are the most common, and the proportion of severe cases caused by EV71 infection is relatively large. Most patients with HFMD have mild symptoms, manifested as fever and rashes or herpes on the hands, feet, and mouth. The course of the disease is self-limiting, and the prognosis is generally good, but a small number of very severe patients may be complicated by asepti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 邓力贾红玲肖传乐刘超武
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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