Kit for human papilloma virus typing detection

A technology of human papillomavirus and kit, which is applied in the direction of microbe-based methods, microbiological measurement/inspection, microbes, etc., which can solve the problems of low HPV sensitivity, high quality requirements for personnel, and low accuracy, and achieve improved coverage , Improve detection specificity, reduce damage effect

Active Publication Date: 2019-12-31
宁波胤瑞生物医学仪器有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Advantages of this method: simple operation, low price, suitable for preliminary screening; disadvantages: hollow cells are the main morphological changes of HPV infection, but due to other viral infections and human factors, vacuoles or hollows may appear in the cells In addition, cytological examination is easily affected by factors such as sampling, staining, and subjective judgment of cytopathologists. Therefore, the application of cytopathological detection of HPV has low sensitivity, poor specificity, high false negative rate and false positive rate, and cannot detect HPV. type
Advantages of this method: simple operation, low price, suitable for preliminary screening; disadvantages: low accuracy, high quality requirements for personnel, severe pain for patients, and unwillingness to cooperate with the inspection due to psychological resistance
Advantages of this method: clear principle and relatively simple operation; disadvantages: the objects of serological detection are antigens and antibodies. Since the human body has a certain hysteresis in the immune response to HPV, serological detection is not suitable for those with no immune response and HPV latent infection. Will produce missed detection
However, there are no kits and detection methods that apply digital PCR technology to HPV16 / 18 / 52 / 5 clinically.

Method used

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  • Kit for human papilloma virus typing detection
  • Kit for human papilloma virus typing detection
  • Kit for human papilloma virus typing detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] 1 Main test equipment:

[0067] Qubit 3.0 Fluorescence Quantitative Instrument: Thermo Fisher;

[0068] Centrifuge: Thermo Fisher;

[0069] PCR amplification instrument: our company's independent research and development company;

[0070] Water bath: Changzhou Zhiborui Instrument Manufacturing Co., Ltd.;

[0071] Biological safety cabinet: Haier Group.

[0072] A kit for detecting human papillomavirus typing, which includes a primer probe premix, a PCR reaction premix, water, a positive quality control, and a negative quality control; the primer probe premix includes detection of 16 , the upstream and downstream primers of HPV types 18, 52, and 58, the fluorescent probes for detecting HPV types 16, 18, 52, and 58, the upstream and downstream primers for detecting human actb gene, and the fluorescent probe for detecting human actb gene; The sequences of each fluorescent probe and upstream and downstream primers in the box are shown in Table 1.

[0073] Table 1 Nucle...

experiment example 1

[0125] Refer to the above detection steps for the standard product, use the sample DNA instead of the plasmid standard product DNA, and detect the HPV infection in the clinical sample; wherein the sample DNA is extracted according to the instructions of the DNA extraction kit used.

[0126] The above kits were used to detect cervical swab samples and urine samples of 50 HPV positive patients. All cervical swab samples had the test results of the qPCR kit as a reference. The test results and analysis results are shown in Table 9 below.

[0127] Table 9

[0128]

[0129]

[0130] The result shows that the kits for qPCR detection of HPV currently used clinically do not provide quantitative detection, but only qualitative typing detection. This is because for cervical swab samples, various factors such as the doctor's sampling method, sampling location, and main infection location will greatly affect the HPV content in the sample. That is to say, the severity of HPV infectio...

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Abstract

The invention belongs to the technical field of diagnostic reagents for human papilloma and particularly relates to a kit for human papilloma virus typing detection and a detection method. The kit comprises a primer probe premixed solution; the primer probe premixed solution comprises upstream and downstream primers for detecting HPV type 16, 18, 52 and 58, fluorescent probes for the HPV type 16,18, 52 and 58 respectively, upstream and downstream primers for detecting human actb genes and fluorescent probes for detecting the human actb genes. For the kit provided by the invention, by adding degenerate bases in a universal primer, the coverage degree of the HPV of different types is increased, and no missed detection is effectively achieved; by adding a design of locked nucleic acid in each fluorescent probe, the detection specificity is improved; on the premise that the reaction is guaranteed to be efficient and accurate, the pore inlet efficiency of a chip is greatly improved; and byusing hot started DNA polymerase and UDG enzyme, the reaction false positive is reduced.

Description

technical field [0001] The invention belongs to the technical field of diagnostic reagents for human papilloma, and in particular relates to a kit and a detection method for typing and detecting human papilloma virus. Background technique [0002] Human papillomavirus (Human Papilloma Virus, HPV) belongs to the papillomavirus genus, is a virus that specifically infects the skin or mucosal stratified epithelium. HPV is a double-stranded DNA virus with a genome size of about 8000 base pairs. [0003] More than 120 types of HPV have been identified, clinically divided into high-risk types and low-risk types according to their cancer-causing risks. Low-risk types are related to sexually transmitted warts or condyloma acuminatum, and generally do not induce cancer. Common low-risk types are 6, 11, 44, 81, etc.; high-risk types are necessary for cervical cancer and uterine intraepithelial neoplasia. Common high-risk types are 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 6...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/6851C12Q1/708C12Q2531/113C12Q2563/107C12Q2545/113
Inventor 刘一博金鑫浩隋硕任鲁风张未来俞育德于军
Owner 宁波胤瑞生物医学仪器有限责任公司
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