Test paper strip/test paper card for rapidly detecting HPV (Human Papilloma Virus) antibody by using double-antibody sandwich method and preparation method thereof

A double-antibody sandwich and antibody technology, applied in biological testing, material testing products, etc., can solve the problem of not seeing high-risk type HPVL1IgG antibody test strips, etc., and achieve improved detection sensitivity and accuracy, low cost, and safety. The effect of high sensitivity

Pending Publication Date: 2018-11-06
陕西医药控股医药研究院有限公司
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Problems solved by technology

[0006] At present, there is no report on the detection of human high-risk HPV L1IgG antibodies, and no test strips (cards) for detecting high-risk HPV L1IgG antibodies by the double-antibody sandwich method based on immunochromatography

Method used

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  • Test paper strip/test paper card for rapidly detecting HPV (Human Papilloma Virus) antibody by using double-antibody sandwich method and preparation method thereof
  • Test paper strip/test paper card for rapidly detecting HPV (Human Papilloma Virus) antibody by using double-antibody sandwich method and preparation method thereof
  • Test paper strip/test paper card for rapidly detecting HPV (Human Papilloma Virus) antibody by using double-antibody sandwich method and preparation method thereof

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Embodiment Construction

[0034] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0035] (1) Preparation of HPV L1 protein and its antibody

[0036] (1) Preparation of HPV L1 protein: use gene cloning technology to construct HPV16L1 whole gene and HPV18L1 whole gene in expression vectors, and use Escherichia coli to express

[0037] 1.1. Amplification primers

[0038]According to the HPV16L1 gene reference sequence (Gene ID: 1489082) and the HPV18L1 gene reference sequence (GeneID: 1489090), HPV L1 gene-specific primers were designed, and the HPV16L1 and HPV18L1 genes were amplified by PCR (the sequence design was completed in May 2016, including restriction endonuclease recognition site):

[0039] HPV16L1 Forward Primer-SphI:5`-CATGCATGC CAGGTGACTTTTATTTACATC-3`

[0040] HPV16L1 Reverse Primer-SacI:5`-CGAGCTC CAGCTTACGTTTTTTGCGTTT-3`

[0041] HPV18L1 Forward Primer-SphI:5`-CATGCATGC GCTTTGTGGCGGCCTAGT-3`

[0042] HPV1...

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Abstract

The invention discloses a test paper strip/test paper card for rapidly detecting an HPV (Human Papilloma Virus) antibody by using a double-antibody sandwich method and a preparation method thereof. The test paper strip (test paper card) comprises a sample pad, a conjugate pad, an analysis membrane and a water sucking pad which are sequentially arranged in a chromatography direction, wherein a nanoparticle marker is arranged on the conjugate pad; a detection zone and a quality control line are arranged on the analysis membrane; the nanoparticle marker is selected from colloidal gold or latex marked anti-human IgG antibody; the detection zone comprises a main HPV capsid protein L1; or the nanoparticle marker is selected from the colloidal gold or latex marked main HPV capsid protein L1; thedetection zone comprises the anti-human IgG antibody. The test paper strip (test paper card) disclosed by the invention is not only convenient and rapid to use and low in cost, but also high in detection accuracy, security and sensitivity, is capable of carrying out on-site rapid detection on HPV16/18 Li IgG antibodies in human bodies, is applicable to detection in hospitals, disease control institutes, basis health units and families, and is applicable to popularization.

Description

technical field [0001] The invention belongs to the field of immune analysis and detection, and in particular relates to a method for rapidly detecting HPV antibodies based on colloidal gold immunochromatography technology and a detection test strip (card). Background technique [0002] Cervical cancer is a common gynecological malignancy. It ranks second in the incidence of female tumors. There are about 470,000 new cases in the world every year and 300,000 deaths. It is second only to breast cancer in women. important cause of death. The etiology of cervical cancer is related to many factors, among which human papillomavirus (human papillomavirus, HPV) infection is the most important factor. According to reports, 99.8% of cervical cancers are combined with HPV infection, while HPV-negative patients hardly occur cervical cancer. At present, there is no effective treatment for HPV infection. Prevention of HPV infection and regular detection of infected people are the main ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 秦湫红高湘肖志强李铮
Owner 陕西医药控股医药研究院有限公司
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