Papillomavirus detection and parting method as well as liquid phase chip thereof

A virus detection, liquid phase chip technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of poor signal-to-noise ratio of detection results, increase coating steps, low fluorescence signal, etc., and achieve improved sensitivity , Increased fluorescence signal value, overcome the effect of low sensitivity

Inactive Publication Date: 2008-08-27
SUREXAM BIO TECH
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  • Abstract
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AI Technical Summary

Problems solved by technology

Due to the need to coat specific oligonucleotide probes on the microspheres, the coating step is increased, and the coated microspheres can only be used for papillomavirus HPV typing
In addition, the existing HPV detection liquid chip technology adopts the method of labeling biotin at the 5' end during PCR amplification, so the fluorescence signal is r

Method used

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  • Papillomavirus detection and parting method as well as liquid phase chip thereof

Examples

Experimental program
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Example Embodiment

[0026] Example 1 The human papillomavirus HPV detection and typing liquid-phase chip kit mainly includes:

[0027] 1. Specific primer sequence (ASPE primer)

[0028]For 22 types of HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 70, 73, 82, 06, 11, 42, 43, 44, respectively Design specific primer sequences (18-22 bp), and add a 24 bp specific tag tag sequence to the 5'end of the primer. These tag tag sequences can respectively complement and pair with the anti-tag sequence on the corresponding microspheres. The ASPE sequence is shown in the following table:

[0029] Table 1 ASPE primer sequence

[0030] SEQ NO.

[0031] 15

[0032] Each ASPE primer consists of two parts, the 5'end is the specific tag sequence for the anti-tag sequence on the corresponding microsphere (as shown in Table 2), and the 3'end is the HPV type-specific primer fragment (as shown in Table 1 above) Shown). All ASPE primers were synthesized by Shanghai Shenggong Biological Eng...

Example Embodiment

[0045] Example 2 Use of HPV detection and typing liquid chip to detect and type clinical samples

[0046] The formulations of the various solutions are as follows:

[0047] 50mM MES buffer (pH5.0) formula (250ml):

[0048] Reagent

[0049] 2×Tm hybridization buffer

[0050] Reagent

[0051] 1MTris-HCl, pH8.0

[0052] Store at 4℃ after filtration

[0053] The biotin-labeled dNTP was purchased from Shanghai Shenggong Biological Engineering Technology Service Co., Ltd.

[0054] 1. Sample selection and sample DNA extraction

[0055] Select 20 cervical samples that were tested and typed for HPV by sequencing. Among them, 8 were low-risk HPV monotype infections, including 3 HPV06 infections and 5 HPV11 infections. Ten were high-risk HPV monotype infections, including 4 HPV16 infections, 4 HPV18 infections, 1 HPV52 infection, and 1 HPV58 infection. The other 2 were mixed infections, one of which was a mixed infection of HPV06 and HPV18, and the other was a m...

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Abstract

The invention discloses papillomavirus detection and a typing liquid chip, and a process which is used to the detection and typing papillomavirus. The liquid chip mainly comprises microsphere which is respectively enveloped with specific anti-tag label sequence and 7-10 T spacer arm sequence which is arranged between anti-tag label sequence and microsphere, wherein anti-tag label sequence is chosen from two or more than two sequences in SEQ ID NO. 25- SEQ ID NO.47, specific ASPE primer is respectively designed aiming at each typing HPV, ASPE primer is chosen from two or more than two sequence from SEQ ID NO.1-SEQ ID NO.23, and primer of target sequence which has mutant sites is amplified. The liquid chip improves the current liquid chip technology, which makes ribonucleotide probe microspheres which is prepared be suitable to different detection projects, largely increases fluorescent signal value which is detected, thereby further increasing sensitivity of the detection, strengthening signal-to-noise ratio, and making detection results more accurate and reliable.

Description

technical field [0001] The invention relates to medical in vitro diagnostic technology, in particular to a method for detecting and typing human papillomavirus and a liquid phase chip thereof. Background technique [0002] Human papilloma virus (HPV) is a small-molecule circular double-stranded DNA virus that spreads widely among people through sexual contact, direct or indirect contact with contaminated objects, and specifically infects human skin and mucous membranes. Squamous epithelial cells can cause a variety of benign or malignant lesions, which seriously threaten human health. According to the sequence variation of HPV gene, it can be divided into many types. There are more than 100 types of HPV identified so far, about 40 of which can infect human reproductive organs, and about 20 are related to tumors. According to the risk of different types of HPV and tumor occurrence, it is divided into high-risk type and low-risk type. Low-risk HPV types such as HPV6, 11, 40,...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 许嘉森林一群杨惠夷
Owner SUREXAM BIO TECH
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