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Assay kit for detecting human papillomavirus and preparation and use thereof

A human papillomavirus, detection kit technology, applied in biochemical equipment and methods, microorganism-based methods, and microbial determination/examination, etc. expensive, etc.

Inactive Publication Date: 2010-10-20
SHANGHAI CENT FOR BIOINFORMATION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing molecular level detection methods are mainly fluorescent quantitative PCR method or the second-generation hybridization capture analysis method patented by Digene Corporation of the United States. The number of subtypes covered by one detection is small, and the former is prone to false positive pollution, while the latter is Expensive and cannot be used for large-scale clinical screening

Method used

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  • Assay kit for detecting human papillomavirus and preparation and use thereof
  • Assay kit for detecting human papillomavirus and preparation and use thereof
  • Assay kit for detecting human papillomavirus and preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Preparation method of human papillomavirus detection kit

[0058] 1.1 Design of primer pairs

[0059] Such as figure 1 As shown in, by detecting the DNA sequence of various subtypes of human papillomavirus, the conserved and variable regions shared by different subtypes of viruses can be found through sequence comparison. Based on these data, the following primer pairs can be designed, and then synthesized Primer sequence (The primer synthesis company is Shanghai Biological Engineering Technology Service Co., Ltd.). The HPV subtype corresponding to each primer pair and the k-mer sequence corresponding to each subtype are shown in the following table:

[0060] 1. Primer pair 1

[0061]

[0062]

[0063] 2. Primer pair 2

[0064]

[0065] 3. Primer pair 3

[0066]

[0067] 4. Primer pair 4

[0068]

[0069] 5. Primer pair 5

[0070]

[0071]

[0072] 6, primer pair 6

[0073]

[0074] 7. Primer pair 7

[0075]

[0076] 8. Primer pair 8

[0077]

[0078] 9. Primer pair 9

[00...

Embodiment 2

[0098] Example 2 High-throughput use of human papillomavirus detection kit:

[0099] Specimen sources: 7 tissue samples from clinical medical units.

[0100] The kit was prepared using the method of Example 1. The difference is that when synthesizing the primers, different DNA tags are added to the 5'ends of the sequencing primers used for detecting different samples. The relationship between the DNA tags and the samples is listed in the following table:

[0101] Sample number

DNA tag sequence

1

CCAATGCA

2

CAATGCAT

3

AATGCATT

4

ATGCATTG

5

TGCATTGG

6

GCATTGGA

7

CATTGGAA

[0102] 2.1 Processing of test specimens:

[0103] 1. Add PBS or saline to the tissue sample and centrifuge to wash;

[0104] 2. Add 20-50uL of digestion lysis solution (mixed solution of 0.5% NP-40 and 0.5% tween-20), and lyse at 95-98 degrees Celsius for 15-30 minutes;

[0105] 3. After lysis, centrifuge at 12000r / min for 5-10min, and take the supernatant for use.

[0106] 2.2 PCR amplification...

Embodiment 3

[0117] Example 3 Clinical sensitivity and specificity test of human papillomavirus detection kit

[0118] 1 Specimen: 50 specimens that have been clinically classified for HPV

[0119] 2 Kit: prepared by the method of Example 1

[0120] 3 Specimen processing, reagent configuration, sample addition and PCR amplification are the same as in Example 2

[0121] 4 Test results:

[0122] The sensitivity of the human papillomavirus detection kit is 94%; the specificity of the human papillomavirus detection kit is 100%; the accuracy of the human papillomavirus detection kit is 96%. The specific test results are shown in the table below

[0123]

[0124]

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Abstract

The invention discloses an assay kit used for detecting human papillomavirus, comprising human papillomavirus sequencing primer solution. The invention also discloses a preparation method and use method of the kit used for detecting high-risk type human papillomavirus. The kit of the invention can determine whether a sample to be detected contains HPV virus and concrete virus subtype. Identity label used for discriminating sample source is added on a DNA sample to be detected, thus meeting high throughout visitation requirement of detecting a plurality of persons at once. The kit of the invention exceeds the traditional detection means, sequence differential is positioned to basic group level, detection is more sensitive and accurate, and meanwhile probability of false positive pollution is avoided.

Description

Technical field [0001] The invention relates to the detection of human papillomavirus, in particular to a detection kit for detecting human papillomavirus and its preparation and use. Background technique [0002] Cervical cancer is a common malignant tumor in gynecology. The mortality rate is second only to breast cancer, and ranks second in women's cancer deaths. According to worldwide statistics, there are about 500,000 new cases of cervical cancer each year, accounting for 5% of all new cases of cancer, 80% of which are in developing countries. There are about 131,500 new cases of cervical cancer in my country each year, accounting for 28.8% of the total number of new cases of cervical cancer in the world. In addition, the number of missed diseases due to conditions is far more than this. The study found that 99.7% of cervical cancer patients can find high-risk HPV infection. The International Agency for Research on Cancer (IARC) clearly stated in 2004 that HPV infection i...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 韦朝春蔡锴晔刘雷高军晖李亦学丁国徽李轩
Owner SHANGHAI CENT FOR BIOINFORMATION TECH
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