Primer, probe and kit for detecting type-16 HPV (human papillomavirus)

A technology for HPV virus and detection primers, which is applied in the direction of microbial determination/inspection, microorganisms, and microorganism-based methods, and can solve problems such as the inability to detect multiple types of HPV viruses at the same time.

Active Publication Date: 2015-11-25
北京鑫诺美迪基因检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In order to solve the problem that multiple types of HPV viruses cannot be detected at the same time in the prior art, the present invention provides detection primers and probes for 16 types of HPV viruses based on fluorescent quantitative PCR technology to achieve rapid, effective and accurate detection of HPV Virus type, used for comprehensive detection of high and low risk types of HPV virus, prediction of HPV virus-related diseases and treatment monitoring

Method used

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  • Primer, probe and kit for detecting type-16 HPV (human papillomavirus)
  • Primer, probe and kit for detecting type-16 HPV (human papillomavirus)
  • Primer, probe and kit for detecting type-16 HPV (human papillomavirus)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0163] Example 1 Construction of HPV quality control

[0164] (1) The 16 types of HPV fragments were ligated to prepare a pseudovirus.

[0165] The 16 types of HPV viruses were respectively amplified with corresponding specific primers (for example, HPV6 type virus template, using the HPV6 type upstream primer HPV6-F and downstream primer HPV6-R designed by the present invention to amplify, collect Product), the amplified products of 16 types of HPV virus were confirmed to be correct by gene sequencing and then packaged into pseudoviruses. The pseudoviruses were prepared and provided by Beijing Boheng Kechuang Biotechnology Co., Ltd.

[0166] (3) Dilute the HPV pseudovirus according to the concentration provided by Beijing Boheng Kechuang Biotechnology Co., Ltd., and dilute it to 1×10 5 copies / mL and 5×10 3 copy / mL, and then detected by ABI7500 fluorescent quantitative PCR instrument.

[0167] The result is as figure 1 Shown: the left is 1×10 detected by 16 kinds of HPV...

Embodiment 2

[0169] Example 2 The composition and use of the detection kit

[0170] 1. Kit composition

[0171] Reagent Element Dosage HPV lysate Anhydrous sodium carbonate, Tris-HCl, EDTA 50μL PCR reaction solution DNA polymerase, dNTPs, Mg 2+ 14.5μL negative control sterile saline

positive control Pseudovirus (1×10 5 copies / mL)

Weak Positive Control Pseudovirus (1×10 3 copies / mL)

Group A Primer Probe Mixture HPV6-F, HPV6-R, HPV6-P; HPV11-F, HPV11-R, HPV11-P 5.5μL Group B Primer Probe Mixture HPV16-F, HPV16-R, HPV16-P; HPV18-F, HPV18-R, HPV18-P 5.5μL Group C Primer Probe Mixture HPV31-F, HPV31-R, HPV31-P; HPV52-F, HPV52-R, HPV52-P 5.5μL Group D Primer Probe Mixture HPV33-F, HPV33-R, HPV33-P; HPV45-F, HPV45-R, HPV45-P 5.5μL Group E Primer Probe Mixture HPV35-F, HPV35-R, HPV35-P; HPV51-F, HPV51-R, HPV51-P 5.5μL Group F Primer Probe Mixture HPV39-F, HPV39-R, HPV39-P; HPV5...

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Abstract

The invention discloses a primer, probe and kit for detecting type-16 HPV (human papillomavirus). The primer and the probe are suitable for screening the HPV related diseases quickly, accurately and simply, do not cross-react with other viruses, are capable of detecting common high and low risk types of HPVs, two types of HPVs and 14 types of HPVs in total, cover a wide range of high risk types of HPVs and are more accurate and complete in detection than kits in the present market. The kit further comprises negative, positive and weakly positive quality controls and helps further increase detection accurateness.

Description

technical field [0001] The invention relates to detection primers, probes and kits for 16 types of HPV viruses. Background technique [0002] Human papillomavirus (HPV) is a small non-enveloped DNA virus belonging to the papillomavirus family. Virus particles are composed of single copy DNA and protein, and have a double-stranded closed-circle DNA genome with a size of about 8000bp. The late region of the late open reading frame (LateRegion, L) and the non-coding long control region (LongControlRegion, LCR). The early region encodes proteins related to HPV replication, transcription, and cell transformation; the late region encodes HPV structural proteins, such as the major capsid protein L1 and the minor capsid protein L2 that make up the viral capsid. The essential difference between the various types of HPV lies in the difference in the nucleotide sequence of the L1 region. [0003] There are more than 100 known HPV types, about 40 of which are related to reproductive ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/708C12Q2531/113C12Q2561/101
Inventor 朱柳王佳刘沛
Owner 北京鑫诺美迪基因检测技术有限公司
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