Unlock instant, AI-driven research and patent intelligence for your innovation.

Kit for detecting five series viruses of infant hand-foot-and-mouth disease and application of kit

A virus detection and kit technology, which is used in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of high reagent price, long detection time, and inability to achieve instant inspection, and achieve good application prospects. , high sensitivity and specificity

Pending Publication Date: 2020-10-09
SHENZHEN PEOPLES HOSPITAL +1
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional fluorescent quantitative PCR method still has high reagent prices, long detection time, and cannot achieve real point-of-care testing (POCT), and cannot meet the requirements of portable, mobile, and on-site rapid inspection.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for detecting five series viruses of infant hand-foot-and-mouth disease and application of kit
  • Kit for detecting five series viruses of infant hand-foot-and-mouth disease and application of kit
  • Kit for detecting five series viruses of infant hand-foot-and-mouth disease and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Example 1 Repeatability Detection

[0108] The positive control of each virus diluted 10 times was used as the sample to be tested for repeated detection, and the detection was repeated 3 times.

[0109] 1. Nucleic acid extraction (used with Hs480 heating centrifuge)

[0110] 1) Take 50 μl of the sample and add 5 μl FastLyseL4, shake and mix;

[0111] 2) Place the mixed sample in the HS480 heating centrifuge machine, heat at 95°C for 2min, and centrifuge at 5000rpm for 2min;

[0112] 3) After centrifugation, take the supernatant as the sample RNA, numbered;

[0113] 4) The positive control, negative control and samples to be tested are processed synchronously.

[0114] 2. Preparation of PCR reagents

[0115] 1) Prepare the PCR reaction solution: pre-mix the ultra-fast PCR buffer1 (68 μl) and the ultra-fast PCR enzyme system (17 μl) (2 more than the actual number of reaction tubes, to avoid loss during sample addition, and to avoid air bubbles during mixing);

[011...

Embodiment 2

[0134] Embodiment 2 specific detection

[0135] Each virus positive control was used as the sample to be tested for specific detection, and the detection method was the same as in Example 1.

[0136] Result: Each virus positive control sample was only positive under the corresponding probe, and the rest were negative, with good specificity. The results were as follows: Figure 1-Figure 5 shown.

Embodiment 3

[0137] Example 3 Sensitivity Detection

[0138] Each virus positive control in the kit (concentration is 1 × 10 5 copies / ml) was serially diluted to 1×10 4 copies / ml, 1×10 3 copies / ml, 1×10 2 copies / ml and 1×10 1 Copies / ml, after preparation, use it as the sample to be tested for sensitivity testing with the corresponding kit.

[0139] Concrete implementation steps are the same as embodiment 1

[0140] Results: The detection limit of CA6 is not higher than 1×10 2 copies / ml, the detection limit of CA10, CA16, EV71 and EVU is not higher than 1×10 1 copies / ml, the result is as Image 6 shown.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of biological detection, and specifically relates to a top-speed detection kit for simultaneously detecting five series viruses of infant hand-foot-and-mouth disease, such as coxsackie virus A6 / A10 / A16 (CA6 / CA10 / CA16), enterovirus 71 (EV 71) and enterovirus universal (EVU), and an application thereof. The kit has high sensitivity and specificity, compared with other fluorescent quantitative PCR, the established method can obtain a result within half an hour, clinical rapid detection can be achieved, real POCT is expected to be achieved, and the kit has good application prospects.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kit for detecting five kinds of children's hand, foot and mouth disease serial viruses and the application thereof. Background technique [0002] Hand-foot-mouth disease is characterized by blisters on the hands, feet, and mouth. The disease is mild and short-lived. It is more common in children and is mainly caused by enterovirus infection. The incubation period is 3 to 5 days, and there are prodromal symptoms such as low fever, general malaise, and abdominal pain. Within 1 to 2 days, painful miliary to mung bean-sized blisters appeared in the oral cavity, pharynx, soft palate, buccal mucosa, tongue, and gums, surrounded by blush, ulcerated into small ulcers, and often salivated and refused to eat due to pain. At the same time, rashes also appeared on the hands and feet. On the dorsal side of the hands and feet, the dorsal edges of the fingers (toes), around the nails, and...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2600/166C12Q2537/143C12Q2563/107C12Q2545/113
Inventor 吴诗品金宁一李体远李昌杜寿文田明尧任琴张星艳王宇航
Owner SHENZHEN PEOPLES HOSPITAL