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Primer combination, probe combination and human papilloma virus nucleic acid detection kit

A technology of human papillomavirus and primer probe, which is applied in the field of probe combination, human papillomavirus nucleic acid detection kit and primer combination, which can solve the problems of high storage temperature requirements, low sensitivity and complicated operation, and achieve storage temperature The effect of low requirements, simple operation and low cost

Pending Publication Date: 2021-03-30
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] 2) High storage temperature requirements
[0022] 3) Complicated operation
[0023] 4) Low sensitivity

Method used

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  • Primer combination, probe combination and human papilloma virus nucleic acid detection kit
  • Primer combination, probe combination and human papilloma virus nucleic acid detection kit
  • Primer combination, probe combination and human papilloma virus nucleic acid detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] 1) Primer probe sequence

[0102] Table 1

[0103]

[0104]

[0105]

[0106] 2) Kit components: ① PCR reaction solution 1: various types of probes and combined probes, various types of primers, Trinice, KOAc, Tween20, glycerol, NaN3, UDG enzyme, rTth, dNTPs, DMSO, ultrapure water. ②PCR reaction solution 2: manganese acetate, magnesium acetate, NaN3, ultrapure water.

[0107] 3) Concentration of kit components: ①PCR reaction solution 1: 0.1-0.5 μM for each type of probe and combined probe, 0.1-0.5 μM for each type of primer, 30-100 mM for Trinice, 100-150 mM for KOAc, 0.005% for Tween 0.1%, glycerol 0.1%~1%, NaN3 0.1%~5%, UDG enzyme 1~10μL, rTth 1~5μL, dNTPs 0.11~0.6mM, DMSO5~50μL / L, ultrapure water. ②PCR reaction solution 2: manganese acetate 1-6mM, magnesium acetate 2-10mM, NaN30.01%-0.1%, ultrapure water.

[0108] Table 2 is the amount of each type of primer probe used in the specific implementation process:

[0109] Table 2

[0110]

[0111]

[0...

Embodiment 2

[0121] Embodiment 2 specificity experiment:

[0122] Use TE solution to dilute the HPV types that are not within the detection range in the human papillomavirus whole genome type reference product to 1×10 4 -1×10 5 Copies / response, for detection, the results are as follows:

[0123] Table 6

[0124]

[0125] Result analysis: There is no cross between this kit and HPV types that are not within the detection range.

Embodiment 3

[0126] Embodiment 3 sensitivity experiment:

[0127] Use TE solution to dilute the HPV types within the detection range in the human papillomavirus whole genome typing reference product to 1×10 3 -1×10 4 Copies / reactions were tested, and the lack of type was detected using a human papillomavirus L1 typing reference product, and the results were as follows:

[0128] Table 7

[0129]

[0130] Result analysis: All HPV types within the detection range were detected, and the Ct value was relatively high, indicating a strong detection ability.

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PUM

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Abstract

The invention discloses a primer combination, a probe combination and a human papilloma virus nucleic acid detection kit. The invention relates to the field of biological detection, in particular to aPCR fluorescent probe detection method for diagnosing high-risk human papilloma virus infection, which comprises the following steps: extracting human papilloma virus nucleic acid in cervical exfoliated cells by using a paramagnetic particle method, and carrying out real-time fluorescence polymerase chain reaction to obtain the high-risk human papilloma virus detection kit to detect 14 HPV DNA types in the sample and typing the HPV 16 / 18; and meanwhile, carrying out female cervical cancer screening in combination with cytological examination. The kit can be used for qualitative detection of high-risk human papilloma viruses in cervical cell samples of patients, and typing identification of HPV16 and HPV18 types is realized while HPV31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68 are detected. The kit is used for qualitatively detecting high-risk HPV DNA in cervical cells of a patient and determining whether the patient needs colposcope examination or not.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a primer combination, a probe combination and a human papillomavirus nucleic acid detection kit. Background technique [0002] Human papillomavirus (HPV) is a kind of non-enveloped double-stranded circular DNA virus with small molecular weight, which obligately infects and parasitizes the epithelial cells of human reproductive organs and other tissues and organs. At present, there are about 150 subtypes of HPV identified, which are divided into low-risk types and high-risk types according to their pathogenicity. Low-risk HPV mainly causes exophytic wart-like lesions on the anus skin, male external genitalia, female labia, urethral meatus, lower vagina, and low-grade cervical intraepithelial neoplasia. High-risk HPV can not only cause external genital warts, but more importantly, external genital cancer, cervical cancer and high-grade cervical intraepithelial neoplasia. The vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/708C12Q1/686C12Q2600/112C12Q2561/101
Inventor 曹雅倩李静静李振红付光宇
Owner AUTOBIO DIAGNOSTICS CO LTD
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