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Kit for PCR (Polymerase Chain Reaction) detection of high-risk human papillomavirus (HPV)

A technology of human papillomavirus and detection kit, which is applied in the determination/inspection of microorganisms, methods based on microorganisms, microorganisms, etc. It can solve the problems of low-load virus detection results that are not very good, and achieve high sensitivity and specificity , the effect of convenient operation

Active Publication Date: 2013-07-03
福州泰普生物科学有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the detection results of low-load viruses are not very good, and special detection and analysis equipment is required

Method used

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  • Kit for PCR (Polymerase Chain Reaction) detection of high-risk human papillomavirus (HPV)
  • Kit for PCR (Polymerase Chain Reaction) detection of high-risk human papillomavirus (HPV)
  • Kit for PCR (Polymerase Chain Reaction) detection of high-risk human papillomavirus (HPV)

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Experimental program
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Embodiment

[0041] One, the composition of kit of the present invention

[0042] The composition of the kit of the present invention is shown in Table 3 and Table 4.

[0043] table 3

[0044]

[0045]

[0046] Preparation before dispensing

[0047] (1)dN(U)TP

[0048] Mix the purchased 100mM dATP, dUTP, dGTP, and dCTP according to 1:2:1:1 (volume ratio). Make 100μL dN(U)TP master solution per tube.

[0049] (2) Primers

[0050] Centrifuge the synthesized primers at 13000 rpm for 1 min, add an appropriate amount of pure water (calculated according to the amount of synthesis) to dissolve the primers to 100 μM, take 0.5 mL centrifuge tubes, and aliquot them into 100 μL primer mother liquors per tube. Before production, H1 and H2 need to add appropriate amount of pure water to make 10μM use solution.

[0051] (3) Probe

[0052] Centrifuge the synthesized probe at 13,000 rpm for 1 min, add an appropriate amount of pure water (calculated based on the amount of synthesis) to dissolv...

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Abstract

The invention aims to design special primers and fluorescent marking probes aiming at gene L1 regions of five high-risk subtypes of high-risk human papillomavirus (HPV) by utilizing a real-time fluorescent PCR (Polymerase Chain Reaction) technology so as to specially amplify and detect gene segments of subtypes of the HPV. A kit for PCR detection of the HPV is provided in the technical scheme of the invention; the kit comprises a DNA (Deoxyribose Nucleic Acid) extraction solution, a PCR reagent, positive quality control serum and negative quality control serum. The PCR reagent contains two amplification primers and five Taman probes. The kit has the beneficial effect that the blank of a fluorescent PCR kit for clinically detecting five high-risk subtypes of the HPV in China is filled up. The sensitivity in detecting samples is up to 1.0*10<3>copies / ml, and when the kit is used for detecting an HPV negative sample and other HPV subtype samples, the detection result is negative, and no cross reaction is caused. The kit has the advantages of high sensitivity and specificity, stability, timeliness, convenience in operation and the like.

Description

technical field [0001] The invention relates to a disease pathogen gene detection technology, in particular to a PCR detection kit for high-risk human papillomavirus. Background technique [0002] The existence of human papillomavirus (human papillomavirus, HPV) is closely related to the benign and malignant hyperplasia of various skin and mucous membranes, especially the occurrence and development of cervical cancer. There is a high correlation between the various gene sequences of the virus and its biological behavior. Different genotypes of HPV have different pathogenic risks. The detection and typing of HPV DNA are of great value for understanding related diseases, judging prognosis and guiding treatment, especially for cancer prediction of female genital tract tumors. Due to the lack of in vitro culture systems and reliable serum reactants, the classification of HPV is almost entirely dependent on its molecular characteristics. By definition, if there is a 10% differe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 林希瑾秦伟胡守旺谢佐福周晓强姚铭锋
Owner 福州泰普生物科学有限公司