Kit for human papilloma virus typing detection

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...

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.

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