Fluorescence PCR method for diagnosis of human papilloma viral infection

A technology of human papillomavirus and fluorescence, which is applied in the direction of material analysis, measuring devices, instruments, etc. through optical means, and can solve problems such as missed detection

Pending Publication Date: 2008-08-27
GENETEL PHARMA SHENZHEN
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, the fluorescent PCR diagnostic products currently on the market to detect low-risk types of HPV can only detect two types (HPV6 / 11, as many as 6 common types), and it is very likely to miss detection clinically

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  • Fluorescence PCR method for diagnosis of human papilloma viral infection
  • Fluorescence PCR method for diagnosis of human papilloma viral infection
  • Fluorescence PCR method for diagnosis of human papilloma viral infection

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Embodiment Construction

[0016] 1. Primer and probe design (see Table 1 and Table 2) and synthesis

[0017] The primers and probes are all commissioned by professional companies to synthesize. The primers are purified by PAGE and the probes are purified by HPLC. The 5'end of the probe is labeled with FAM fluorescent group, and the 3'end is bound with MGB and NFQ quenching groups.

[0018] Table 1. Sequence list of primers and TaqMan-MGB probe oligonucleotides for HPV detection

[0019] Serial number

Sequence name

Oligonucleotide sequence (5′-3′)

Base length (bp)

SEQ ID NO.1

HPV6 probe

ATATCGCCAT CCTGTATAAC A

21

SEQ ID NO.2

HPV11 probe

TGTCCCCATC CTGTATAA

18

SEQ ID NO.3

HPV40 probe

TrCAGGATGG CGACATG

17

SEQ ID NO.4

HPV42 probe

ATATCCCCAT CCTGAATAA

19

SEQ ID NO.5

HPV43 probe

CATGTCACCA TCCTGTATAA

20

SEQ ID NO.6

HPV44 probe

CACCATCCTC AATTACACTA G

21

SEQ ID...

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Abstract

The present invention relates to a fluorescent PCR (polymerase chain reaction) detection method for diagnosing low risk type human papilloma virus (HPV) infection, belonging to the in vitro nucleic acid diagnosis field. The present invention comprises a multiple polymerase chain reaction (PCR) system based on fluorescent PCR technique, includes a forward primer and a reverse primer which aim at a plurality of HPV types, as well as a fluorescent probe, and can simultaneously detect DNA comprising six HPV types at most, namely HPV6, 11, 40, 42, 43 and 44, in one reaction tube under proper PCR conditions. The detection method can conveniently and rapidly diagnoss HPV infection in clinical samples, is high in specificity, and has great clinical value for preventing and controlling condyloma acuminatum in an early stage, blocking infection sources and reducing HPV infection.

Description

Technical field [0001] The invention belongs to the field of in vitro nucleic acid diagnosis, and is a fluorescent polymerase chain reaction (PCR) method for simultaneously detecting one or more human papillomavirus (HPV) infections in clinical samples. Background technique [0002] Condyloma acuminata (Condyloma acuminata; Condyloma acuminatum; Genital warts) is an epidermal tumor-like hyperplasia caused by human papillomavirus (HPV) infection. The virus replicates on the basal layer of the infected skin and mucosal tissues, and induces epithelial proliferation, destroys cell growth regulation, and causes local proliferative lesions. [0003] Papillomavirus belongs to the papillomavirus genus of the Papillomavirus family, which includes a variety of animal papillomaviruses and human papillomavirus (Human Papillomavirus, HPV). Among them, human papillomavirus (HPV) is a common DNA virus that specifically infects human epidermis and mucosal squamous epithelium. It can be transmitt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/00C12Q1/68
Inventor 杨梦甦黄鹤黄明辉柏干荣
Owner GENETEL PHARMA SHENZHEN
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