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A method for detection of human papillomavirus (HPV) type

一种人乳头瘤病毒、类似物的技术，应用在寡核苷酸序列及其诊断试剂盒领域，能够解决携带污染等问题

Inactive Publication Date: 2012-05-16
ALLUM JENKINS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This includes the risk of carry-over contamination

Method used

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  • A method for detection of human papillomavirus (HPV) type
  • A method for detection of human papillomavirus (HPV) type
  • A method for detection of human papillomavirus (HPV) type

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0094] Example 1- plasmid

[0095] Plasmids comprising the target regions of the L1 genes of HPV6, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 66 were transformed into Escherichia coli DH5-α, and isolated by cesium chloride density gradient centrifugation (Fritsch et al.) or Qiagen HiSpeed ​​Midi kit (Qiagen GmbH, Hilden, Germany). DNA concentration was determined by UV spectrophotometry.

Embodiment 2

[0096] Example 2- clinical samples

[0097] Clinical samples were cervical cytobrush samples sent to our laboratory for routine HPV analysis for secondary screening for low-grade proliferation (LSIL) or suspicious cytology (ASCUS or inadequate samples). Samples were collected and transferred in CYTYC thin prep transfer medium (medium CYTYC, Crawley, UK) according to the manufacturer's recommendations. 10 ml samples were centrifuged at 3000 rpm for 10 minutes and the pellet was resuspended in 100 μl of phosphate buffered saline, after which DNA was extracted using the MagNAPure Automated DNA Extractor and the MagNAPure LC DNA Extraction Kit (Roche Diagnostics, Penzberg, Germany). 100 μl of eluted DNA. The test was evaluated using clinical samples previously tested with the HCII assay and typed using universal PCR and reverse line blot.

Embodiment 3

[0098] Example 3- hybrid capture test

[0099] The Digene hybrid capture assay (Digene, Gaithersburg, MD) was performed using 25 ml samples according to the manufacturer's instructions.

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Abstract

The present invention describes a method for detection of human papillomavirus (HPV) types and a kit for detection of said HPV types.

Description

technical field [0001] The invention relates to the detection, typing and quantification of human papillomavirus (HPV) in clinical samples. More specifically, it relates to oligonucleotide sequences capable of being used in amplification reactions and used for simultaneous detection, typing and quantification of multiple HPV genomes and diagnostic kits thereof. Background technique [0002] In the 1980s, human papillomavirus (HPV) infection was reported as the cause of cervical cancer. The relationship between cancer malignancy and HPV genotype was found to be significant. More than 120 HPV types have since been identified. Thirteen HPV types have been identified as tumorigenic in mucosal epithelium. These types include HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 66. [0003] Human papillomavirus infection is confined to the epithelium, limiting its access to immune organs and it is poorly immunogenic. Therefore, serology is not a valid test for HPV. Likewis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/70C12N15/11
CPCC12Q1/6851C12Q1/708
Inventor 安德鲁·詹金斯安妮-格瑞·阿勒姆琳达·斯特兰德
Owner ALLUM JENKINS
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