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Kit for detecting high-risk human papillomavirus and its preparation and application

A technology of human papillomavirus and kit, which is applied in the field of diagnostic reagents for human papillomavirus, and can solve the problems of high false positive rate and easy alternate contamination

Active Publication Date: 2011-11-30
SHANGHAI ZJ BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to multiple amplifications involved, it is easy to have alternate contamination, resulting in a high false positive rate

Method used

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  • Kit for detecting high-risk human papillomavirus and its preparation and application
  • Kit for detecting high-risk human papillomavirus and its preparation and application
  • Kit for detecting high-risk human papillomavirus and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Example 1 The preparation method of the human papillomavirus (HPV) high-risk type typing detection kit detection mixed solution:

[0104] 1. Synthesize oligonucleotide probes according to the following sequence:

[0105]

[0106]

[0107] 2. Preparation of nucleic acid fluorescent PCR detection mixture:

[0108] (1), preparation of HPV16 type and HPV 56 type nucleic acid fluorescent PCR detection mixture:

[0109] Human papillomavirus (HPV) type 16 upstream primer 1.2 μl / test; downstream primer 1.2 μl / test; probe 0.2 μl / test; human papillomavirus (HPV) type 56 upstream primer 1.2 μl / test; downstream primer 1.2 μl / test; probe 0.2μl / test; PCRMIX-20μl / test; deionized water 10.4μl / test and mix well.

[0110] (2), preparation of HPV18 type and HPV 45 type nucleic acid fluorescent PCR detection mixed solution:

[0111] Human papillomavirus (HPV) type 18 upstream primer 1.2 μl / test; downstream primer 1.2 μl / test; probe 0.2 μl / test; human papillomavirus (HPV) type 45 u...

Embodiment 2

[0124] The use of embodiment 2 human papillomavirus (HPV) high-risk typing detection kit:

[0125] Source of specimens: The specimens used come from clinical medical units.

[0126] 2.1 Processing of test specimens:

[0127] Add 1ml of sterile saline to the specimen, shake well, absorb the liquid and transfer it to a 1.5ml centrifuge tube, and centrifuge at 13,000rpm for 5 minutes. Add 1 ml of sterile saline to the precipitate, mix well, centrifuge at 13,000 rpm for 5 minutes, and repeat the washing once. The precipitate was directly added to 50 μl of nucleic acid extraction solution and mixed thoroughly, bathed in boiling water for 10 minutes, then centrifuged at 13,000 rpm for 5 minutes, and 4 μl of the supernatant was taken as a PCR reaction template.

[0128] 2.2 Reagent preparation:

[0129] (1) Take 36 μl HPV16 and HPV 56 nucleic acid fluorescent PCR detection mixture and 0.4 μl Taq enzyme, shake and mix for a few seconds, and centrifuge at 3000 rpm for a few seconds....

Embodiment 3

[0154] Example 3 Human papillomavirus (HPV) high-risk type typing detection kit clinical sensitivity and specificity test:

[0155] 3.1 Specimen: 50 specimens that have been clinically classified into HPV high-risk types

[0156] 3.2 Sample processing, reagent configuration, sample loading and PCR amplification are the same as Example 1

[0157] 3.3 Test results:

[0158] Among the 50 specimens, the test results of the high-risk HPV typing detection kit and the clinical unit were both positive in 42 cases, and the sensitivity of the high-risk HPV typing detection kit was 97.67% ; Human papillomavirus (HPV) high-risk type detection kit detection results and clinical unit detection results were negative in 7 cases, and the specificity of human papillomavirus (HPV) high-risk type detection kit was 100%; The accuracy of the HPV high-risk typing detection kit is 98%. The specific test results are shown in the table below.

[0159] Test results

[0160] Specimen number Test res...

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Abstract

The invention relates to an auxiliary diagnostic kit for cervical cancer and high-grade intraepithelial lesions of the cervix, and discloses a kit for detecting high-risk human papillomaviruses, including high-risk HPV nucleic acid fluorescent PCR detection mixture and Taq enzyme. The invention also discloses the preparation method and application method of the kit for detecting high-risk human papillomavirus. The kit of the invention can be used to detect high-risk human papillomaviruses, and overcomes the problems of high false positive rate, low specificity, high cost and the like in prior art for detection of high-risk human papillomavirus (HPV) typing.

Description

technical field [0001] The invention relates to a diagnostic reagent for human papillomavirus (HPV), in particular to a kit for detecting high-risk human papillomavirus and its preparation and application. Background technique [0002] HPV is the English abbreviation of human papillomavirus, which is a general term for a group of viruses, forming a family, and their virus shapes are similar. At present, there are more than 80 HPV types that have been identified. According to the risk of different types of HPV and tumor occurrence, they are divided into low-risk types and high-risk HPV types. High-risk HPV types include HPV16, 18, 31, and 33. , 35, 39, 45, 51, 52, 56, 58, 59, 68, a total of 13 types, which are related to the occurrence of cervical cancer and cervical intraepithelial high-grade lesions. A large number of studies on the etiology of cervical cancer have confirmed that high-risk HPV infection is a necessary factor for the occurrence of cervical cancer. The pers...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64C12R1/93
Inventor 朱旭平朱勤玮马丽丽
Owner SHANGHAI ZJ BIO TECH
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