EV71 virus IgA antibody detection test strip and application thereof

An antibody detection and test strip technology, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of false negatives, long detection time, and high requirements for experimental conditions, and achieves the goal of reducing false negatives, improving reliability, and simple operation. Effect

Active Publication Date: 2017-05-10
GUANGZHOU RHFAY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Cell isolation and culture is the gold standard method for determining EV71 virus infection, but its clinical diagnostic application is limited by the shortcomings of long detection time (more than 5 days), high requirements for experimental conditions, and high technical requirements for operators.
[0005] RT-PCR nucleic acid molecular detection is the most sensitive method among the early detection methods of EV71 virus. To judge the infection status by detecting EV71 virus in throat swabs and anal swabs of patients, professional technicians and expensive fluorescent PCR amplification instruments are required. It is difficult for primary medical departments to carry out this diagnostic method
In addition, when collecting cells from the patient's throat, it is more likely to cause the patient's discomfort, regurgitation or vomiting, which makes the work of medical staff more difficult, and also easily increases the risk of virus contamination in the sampling

Method used

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  • EV71 virus IgA antibody detection test strip and application thereof
  • EV71 virus IgA antibody detection test strip and application thereof
  • EV71 virus IgA antibody detection test strip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] [Example 1] EV71 virus IgA antibody detection test strip preparation (capture method)

[0029] 1. Source of raw materials: Goat anti-human IgA antibody was purchased from Sigma Company, and human IgA antibody was purchased from Guangzhou Nuosi Biotechnology Co., Ltd. The sequence of the polypeptide fragment of the EV71 virus fusion protein antigen is shown in Seq ID NO.1, a total of 58 amino acids, entrusted to Shanghai Gil Biochemical Company to be responsible for the synthesis, with a purity greater than 98%, and freeze-dried for storage. Colloidal gold solution (40nm) was purchased from Shanghai Jieyi Biotechnology Co., Ltd.

[0030] 2. Pretreatment of sample pad and marker pad

[0031] Both the sample pad and the marker pad are made of polyester film, which need to be soaked with pretreatment solution before use. The pretreatment solution is composed of 0.01-0.02mol / L phosphate buffer (pH7.0-7.5), 3g / 100ml-6g / 100ml non-ionic surfactant, 1ml / 100ml-4ml / 100ml Tween a...

Embodiment 2

[0039] [Example 2] EV71 virus IgA antibody detection test strip preparation (indirect method)

[0040] 1. Source of raw materials: Goat anti-human IgA antibody was purchased from Sigma Company, and EV71 virus VP1 protein antibody was purchased from Guangzhou Nuosi Biotechnology Co., Ltd. The sequence of the polypeptide fragment of the EV71 virus fusion protein antigen is Seq ID NO.1, a total of 58 amino acids, entrusted to Shanghai Gil Biochemical Company to be responsible for the synthesis, with a purity greater than 98%, and freeze-dried for storage. Colloidal gold solution (40nm) was purchased from Shanghai Jieyi Biotechnology Co., Ltd.

[0041] 2. Pretreatment of sample pad and marker pad

[0042] Both the sample pad and the marker pad are made of polyester film, which need to be soaked with pretreatment solution before use. The pretreatment solution is composed of 0.01-0.02mol / L phosphate buffer (pH7.0-7.5), 3g / 100ml-6g / 100ml non-ionic surfactant, 1ml / 100ml-4ml / 100ml Tw...

Embodiment 3

[0050] [Example 3] Detection of EV71 virus IgA antibody (capture method)

[0051] Please refer to the attached figure 2 , with figure 2 Schematic diagram of the method for releasing a saliva sample from a saliva collection stick. The present invention absorbs saliva samples in the patient's mouth through the saliva collection stick 22, and the aforementioned collection stick is placed in a sample diluent 23 containing 1-2ml (containing 0.5% bovine serum albumin, 0.2% casein, 1% tryptone and 0.02 % sodium azide PBS buffer solution) in the sample tube 21, fully stir, squeeze the cotton swab head liquid of the collection stick, and discard the collection stick.

[0052] Please refer to the attached image 3 , image 3 It is a schematic diagram of the detection method of the EV71 virus IgA antibody detection test strip of the present invention. Get the EV71 virus IgA antibody detection test strip prepared by [Example 1] and put it into the above-mentioned sample tube 21 equ...

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Abstract

The invention relates to an EV71 virus IgA antibody detection test strip which comprises a sample pad, a label pad, a nitrocellulose membrane, absorbing paper and a back plate. The nitrocellulose membrane is provided with an EV71 virus VP1 protein antigen peptide fragment detection line and a human IgA antibody control line. The label pad is provided with an an-human IgA anti-body-colloidal gold marker. The EV71 virus IgA antibody detection test strip is used for saliva EV71 virus detection; by detecting an IgA anti-body in a saliva sample, EV71 viruses can be quickly screened, reliability of detection results can be improved, and false-negative conditions are reduced. A detection method is simple to operate, the results are quick, the collection of a detected sample has no invasiveness, and discomfort and cross infection caused by conventional blood sampling and collection of throat swab, anus swab samples and cerebrospinal fluid are avoided.

Description

technical field [0001] The invention relates to a virus detection test strip and its application, in particular to a EV71 virus IgA antibody detection test strip and its application. Background technique [0002] Enterovirus 71 (referred to as EV71 virus) belongs to the family Picornaviridae Enterovirus, a non-enveloped single-stranded positive-sense RNA virus, and is one of the main pathogens causing hand-foot-mouth disease (HFMD). one. The EV71 virus can be transmitted through air droplets and other channels, and is highly pathogenic. It often causes severe central nervous system complications such as aseptic meningitis, encephalitis, poliomyelitis-like paralysis, and neurogenic pulmonary edema. Cause death, currently there is no effective treatment, mainly through early diagnosis and symptomatic treatment. EV71 virus mainly infects children under the age of 6, especially children aged 2-5. Most children in this age group attend kindergartens. Therefore, kindergartens ar...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56916G01N33/577
Inventor 游爱萍李有生刘轶锴向怀勇
Owner GUANGZHOU RHFAY BIOTECH CO LTD
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