Composite amplification system for nine-valent HPV virus typing detection and kit for composite amplification system

A compound amplification system and HPV virus technology, which is applied in the field of compound amplification system and its kit, can solve the problem of HPVL1 gene deletion and other problems, and achieve the effect of stable signal intensity, strong system specificity and good stability

Inactive Publication Date: 2020-05-01
SUZHOU MICROREAD GENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the L1 region sequence is very conservative, which is the main basis for distinguishing different subtypes, but there is a certain probability that the HPV L1 gene will be deleted during the process of HPV viral DNA integration into the human genome

Method used

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  • Composite amplification system for nine-valent HPV virus typing detection and kit for composite amplification system
  • Composite amplification system for nine-valent HPV virus typing detection and kit for composite amplification system
  • Composite amplification system for nine-valent HPV virus typing detection and kit for composite amplification system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Detection of cervical smear samples

[0055] Collect cervical smear samples, use the kit method, phenol chloroform extraction method, Chelex-100 method or magnetic bead method to extract DNA, amplify the DNA according to the above system and amplification procedure, and then the amplified product is subjected to capillary electrophoresis and analyzed using ABI A series of genetic analyzers were used for testing.

[0056] The specific operation of the capillary electrophoresis detection method is as follows: 1 μL amplification product + 0.5 μL molecular weight internal standard QD550 + 8.5 μ deionized formamide (HIDI), briefly centrifuged after mixing, denatured at 95 ° C for 3 minutes, immediately ice bathed for 3 minutes, and used after centrifugation ABI series genetic analyzer electrophoresis detection.

[0057] Finally, the detection results were carried out on data analysis software such as Genemapper. Test results such as figure 1 , it can be seen fr...

Embodiment 2

[0058] Embodiment 2: detection of national standard product

[0059] The national reference material for human papillomavirus whole genome typing (the standard contains the β-globin detection site sequence), from China National Institutes for Food and Drug Control, batch number: 360003-201602. In addition to the 9 HPV types contained in the detection kit, HPV67, HPV81 and human genomic DNA F312 were randomly selected as templates for amplification detection. Test results such as figure 2 , image 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , it can be seen from the figure that the test results of the 9 HPV standard products contained in the kit are correct, and there is no non-specific occurrence, and the typing results of the L1 area and the E area are consistent. However, in the detection results of HPV67, HPV81 standard products and F312, only β-globin was detected, and no other types and non-specific peaks appeared.

Embodiment 3

[0060] Embodiment 3: typing standard

[0061] The 9 kinds of HPV typing standards contained in the equal ratio mixing kit are mixed and diluted according to the copy number provided by the standard, and the copy number of each HPV typing in the diluted mixture is below 100 copies / μL. According to the present invention, the kit amplification system and program amplification are provided, the template addition amount is 1 μL, and the detection results after capillary detection are as follows: Figure 8 . It can be seen from the figure that both the L1 and E regions of the nine HPV types were detected.

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Abstract

The invention discloses a composite amplification system for nine-valent HPV virus typing detection and a kit for the composite amplification system. The amplification system includes 19 pairs of detection primers with nucleotide sequences as shown as SEQ ID NO.1-38. By adopting the amplification system and method disclosed by the invention, nine HPV types can be detected, the selected type is a type corresponding to a nine-valent vaccine as well as is high in degree and more practical in detection significance. Each detectable type has two detection sites which are set in an L1 region and anE region of an HPV genome respectively, so that the kit can be ensured to detect the HPV types more efficiently, accurately and sensitively.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a composite amplification system and a reagent kit for non-type detection of nine-valent HPV viruses. Background technique [0002] Human papillomavirus (Human Papilloma Virus, HPV for short) is a papillomavirus A genus of the Papovaviridae family. It is a sexually transmitted disease caused by spherical DNA virus infection. The main infection areas are human epidermis and mucous membranes. Squamous epithelium, HPV is known to cause benign tumors and warts in humans. The virus is highly specific, and more than 130 types have been isolated so far. The main types are HPV1, 2, 6, 11, 16, 18, 31, 33, and 35, among which HPV16 and 18 may be associated with cervical cancer in women for a long time. related. It takes a long time from HPV infection to cervical cancer, so detecting HPV infection as early as possible is conducive to preventing the occurrence of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/686C12Q1/708C12Q2600/166C12Q2563/107
Inventor 于在亮郭丹高运通李思思
Owner SUZHOU MICROREAD GENETICS
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