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Primer and probe for detection of human papillomavirus, and detection method of human papillomavirus using same

A technology for virus detection and papilloma, applied in biochemical equipment and methods, microbiological measurement/testing, DNA/RNA fragments, etc., can solve problems such as difficulty in accurate confirmation, reduced sensitivity, and clinical use restrictions, and achieve effective diagnosis , the effect of reducing time and cost

Inactive Publication Date: 2012-04-25
THE CATHOLIC UNIV OF KOREA IND ACADEMIC COOPERATION FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] However, when the above-mentioned traditional techniques are compared with the various HPV types disclosed so far, they can only diagnose extremely limited types of HPV, and there are problems such as a significant reduction in the sensitivity of detection based on HPV types, which are limited in actual clinical use. larger limit
In particular, even if the amplification product is obtained by PCR reaction using universal primers, it is difficult to accurately confirm whether the newly identified and classified HPV type is a high-risk group or a low-risk group. In the case of using type-specific primers, in order to confirm whether In the presence of HPV, there is the trouble of using various primers at the same time, which limits the wide use in clinical practice

Method used

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  • Primer and probe for detection of human papillomavirus, and detection method of human papillomavirus using same
  • Primer and probe for detection of human papillomavirus, and detection method of human papillomavirus using same
  • Primer and probe for detection of human papillomavirus, and detection method of human papillomavirus using same

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Effect test

Embodiment 1

[0086] Design and synthesis of primers for amplification of 6 subtype-specific DNA base sequences

[0087] The inventors of the present invention designed and synthesized PCR primers for detection as specific base sequences for each type of HPV gene (intergenic sequence) as shown in Table 1, which can prevent HPV- 1. The mutual cross-reaction of HPV-2, HPV-3, HPV-4, HPV-27 and HPV-57 can be detected at one time.

[0088] 【Table 1】

[0089]

Embodiment 2

[0091] Preparation of kits for detection of human papillomavirus

[0092] The inventors of the present invention prepared a multiplex polymerase chain reaction mixture comprising 6 pairs of PCR primers (sequence number (1-12)) synthesized in Example 1, and prepared a detection kit. Among them, the detection The kit consists of 10X reaction buffer, multiple primer mixture (sequence number 1-12) and Taq DNA polymerase (5U / μl).

Embodiment 3

[0094] Multiplex PCR using primer pairs for each of the 6 subtypes

[0095] The inventors of the present invention performed a multiplex polymerase chain reaction using standard HPV DNAs of six subtypes and amplification primer pairs for each subtype.

[0096] First, wart skin tissue is collected, treated with proteinase K, and DNA is extracted from the tissue, which is then used as template DNA to perform multiple polymerase chain reaction using PCR primer mixture. If the multiplex polymerase chain reaction is over, the amplified DNA is electrophoresed on a 1.5% agarose gel, and the subtype is confirmed and determined according to the presence or absence of specific DNA base sequence amplification and its size. The results are as follows figure 1 shown.

[0097]From the results, it can be confirmed that the first primer pair consisting of the nucleotide sequences of SEQ ID NO: 1 and 2 amplifies HPV-1 DNA, and the second primer pair consisting of the nucleotide sequences of...

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Abstract

The present invention relates to a primer and a probe for detecting human papillomavirus, and a detection method of human papillomavirus using same. More specifically, the present invention relates to a primer for specifically detecting human papillomavirus which comprises at least one primer pair selected from the group consisting of a first primer pair composed of the base sequence of sequence numbers 1 and 2; a second primer pair composed of the base sequence of sequence numbers 3 and 4; a third primer pair composed of the base sequence of sequence numbers 5 and 6; a fourth primer pair composed of the base sequence of sequence numbers 7 and 8; a fifth primer pair composed of the base sequence of sequence numbers 9 and 10; and a sixth primer pair composed of the base sequence of sequence numbers 11 and 12, a probe, a microarray for detecting human papillomavirus containing the same, a kit for detecting human papillomavirus, and the detection method of human papillomavirus using the primer or the probe. The primer for detecting human papillomavirus according to the present invention and the detection method of human papillomavirus using the same are very useful for specifically, rapidly and precisely detecting the presence or absence of and type of HPV-1, 2, 3, 4, 27 and 57 which are the human papillomaviruses that cause mucocutaneous warts.

Description

technical field [0001] The present invention relates to a primer and a probe for detecting human papillomavirus and a method for detecting human papillomavirus using the same. Background technique [0002] Human papilloma virus (HPV) is a circular double-helix DNA virus belonging to the Papovaviridae family, which can infect human and mammalian epithelial cells, and cause various malignant tumors such as warts. Symptoms are closely related and are known to be important etiological factors of cervical cancer. [0003] Among the more than 100 known HPV species so far, more than 40 species have been found in reproductive organs, and it has been confirmed that the development speed of the high-risk groups HPV-16 and HPV-18 is about 40% faster than other HPV types, and other HPV types Significance accounts for approximately 70% of all HPVs found in cervical cancer. In addition, HPV-26, HPV-30, HPV-31, HPV-33, HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, HPV-53, HPV-56, HPV-58, HPV-...

Claims

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Application Information

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IPC IPC(8): C12N15/11
CPCC12Q1/686C12Q1/708
Inventor 崔顺荣朴荣珉金范俊
Owner THE CATHOLIC UNIV OF KOREA IND ACADEMIC COOPERATION FOUND
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