To date, HPV is difficult to detect by traditional viral culture and serological techniques, the main experimental diagnostic techniques are nucleic acid hybridization.
1, histopathological examination: visible keratinized cells and concave hollow cells. Accuracy 80-90%.
2, anti-HPV coat antigen immunohistochemistry: with HPV immunization of animals produced by polyclonal antibodies to check tissue HPV antigen. The peroxidase-antiperoxidase method (i.e., PAP) is commonly used to show the viral proteins within the warts to prove that there are viral antigens in the wart damages.When the HPV proteins are positive, there can be a weak reddish-positive reaction within the superficial epithelial cells of the warts.
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The HPV antigen is the most common antigen in the world.) There are also PAAP and ABC methods. Seeing brown granular deposits in the nucleus of vacuolated cells is positive; the detection rate is low, the sensitivity is not high, and it cannot be typed, with a positive rate of 40-60%.
3, genetic diagnosis:
3.1, poly and enzyme chain reaction: the development of the PCR method has the advantages of specificity, sensitivity, simplicity, rapidity and so on, opening up a new way for HPV detection. Detection of HPV by PCR is superior to nucleic acid hybridization. It is highly sensitive, and if the results are analyzed directly by gel electrophoresis in the GP-PCR method, 200 copies of HPVDNA can be detected in the specimen, and if the PCR product is detected by nucleic acid hybridization, the sensitivity is improved and 10 copies of HPVDNA can be detected.
Scraping boards or saline moistened cotton swabs are used to collect secretions and cells from the vagina and the external osphere of the uterine cervix. While performing cytology, the specimens were placed in 5 ml of PBS containing 0.05% thimerosal, washed twice by centrifugation (3000 g, 10 min) with PBS, and the deposited cells were resuspended in 1 ml of PBS, and 0.5 ml of the cell suspension was taken to extract the DNA.
PCR assay showed a specific HPV-DNA amplification zone. In view of the high sensitivity of PCR technology, the use of genital tract exfoliated cells as the test material is sufficient to meet the test requirements, avoiding the biopsy of the material, grinding tissue cumbersome operation. In general, the PCR amplification product is subjected to gel electrophoresis, and the diagnosis can be made directly by observing the resulting DNA. Therefore, PCR technology to detect HPV experimental cycle is short, simple and rapid. The disadvantage is that it can not be localized, do not know whether the virus is alive or dead. False positives can easily occur. Positivity can persist for a considerable period of time after healing. Specificity also decreases gradually.
3.2, DNA hybridization: HPVDNA molecular hybridization techniques. Such as PCK method. EGFR immunostaining method. CP?14 (Leu?M3) immunostaining method (fluorescence microscope 400 times can see more than 3 fluorescent dots as positive). Nucleic acid hybridization can detect HPV-DNA-related sequences, and specific HPV-DNA amplification bands can be detected by PCR. However, it is cumbersome and expensive. Requires certain equipment and conditions.
4, histochemical examination: take a small amount of lesions made of smear, with specific anti-Human Papilloma Virus antibody for staining. If there is a viral antigen in the lesion, the antigen and antibody bind. In the peroxidase-antiperoxidase (PAP) method, the nuclei are stained red. This method is specific and rapid and is helpful in diagnosis.
5, cytology test: with - warts tissue smear for Pap staining, visible two kinds of cells, namely vacuolated cells and keratinized cells. It can be diagnosed.
6, local experimental method: toluidine blue test: 1% toluidine blue coated warts or normal skin around the drying with 1% acetic acid decolorization, there is still staining for positive. See tumor or acromegaly patients.
7, transmission electron microscopy test: keratinocyte positive rate bottom. 25-45%. Can be combined with colposcopy, magnification examination.
8, colposcopy: must be assisted by acetic acid white test. As long as the regular medical institutions testing, generally more accurate.