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Method, kit and probe assembly for virus genotyping detection

A virus gene and detection probe technology, applied in the field of virus genotyping detection, can solve the problems of high price, cumbersome method operation, false positives, etc., and achieve high specificity, low false positive rate, and accurate and reliable detection results. Effect

Active Publication Date: 2017-02-15
GUANGZHOU EZLIFE BIO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the HC2 detection method is expensive, and the method is cumbersome to operate, and it is easy to cross-contaminate and cause false positives during the operation.

Method used

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  • Method, kit and probe assembly for virus genotyping detection
  • Method, kit and probe assembly for virus genotyping detection
  • Method, kit and probe assembly for virus genotyping detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] This embodiment takes the detection of HPV genotype as an example to describe the kit for virus genotyping detection provided in this embodiment. In this embodiment, the HPV subtypes are detected from cells at the cervix of two different patients with cervical cancer (i.e., samples to be tested), and the samples to be tested from two different patients are named as sample 1 and sample 2, respectively. Sample 1 and sample 2 were tested at the same time, and samples 1 and 2 were taken and stored at -20°C.

[0070] The kit for virus genotyping detection provided by this embodiment includes a probe combination, which includes 16 probe pairs for HPV subtype detection, and the capture probe and detection probe of each probe pair are as follows: The solution forms exist independently, and the final concentration of the probes is 1 μM. The 16 probe pairs are respectively: the first probe pair for detecting HPV 16 subtypes, the base sequence of its capture probe is shown in SEQ...

Embodiment 2

[0129] The kit used for virus genotyping detection provided by this embodiment includes the same probe combination (same as Example 1), immobilizer, streptavidin-labeled horseradish peroxidase (existing in solution form) and Its substrate and immobilizer are conductive polymer and ionic compound, wherein the conductive polymer is pyrrole and the ionic compound is potassium chloride. The substrate is a solution containing TMB. All the other are with embodiment 1.

[0130] In this embodiment, the detection group is set to 1, named as 18 subtype simultaneous detection groups, and the detection group includes a positive control well, a negative control well (repeated 4 times), a detection well of sample 1 and a sample 2 detection holes. 16 kinds of probe pairs described in Example 1 were added to each well, and 18 kinds of HPV subtypes were detected simultaneously). All the other are with embodiment 1. If there is a positive detection result, the detection result can indicate ...

Embodiment 3

[0133] The kit used for virus genotyping detection provided by this embodiment includes the same probe combination (same as Example 1), immobilizer, streptavidin-labeled horseradish peroxidase (existing in solution form) and Its substrate, cleaning solution.

[0134] The immobilizer is a conductive polymer and an ionic compound, wherein the conductive polymer is pyrrole and the ionic compound is potassium chloride. The substrate is a solution containing TMB. The washing solution includes washing solution A and washing solution B, wherein washing solution A is SSC buffer containing SDS, and washing solution B is PBS buffer containing Tween20. All the other are with embodiment 1.

[0135] In this embodiment, the detection group is set to 16 groups, which are 16 subtype groups, 18 subtype groups, 26 subtype groups, 31 subtype groups, 35 subtype groups, 39 subtype groups, and 45 subtype groups , 51 subtype group, 52 subtype group, 53 subtype group, 56 subtype group, 59 subtype ...

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Abstract

The invention relates to the technical field of gene detection and discloses a probe assembly for virus genotyping detection. The probe assembly comprises at least one probe pair, each probe pair comprises a capture probe and a detection probe, each capture probe is used for combining with a target sequence, and the detection probes correspond to the capture probes and can be combined with the target sequences. An affinant for combination with catalyzing enzymes is marked at a 3'-end or a 5'-end of each detection probe. The probe assembly can be used for genotyping detection of viruses in to-be-detected samples and is high in specificity and sensitivity and low in false positive rate, and detection results are accurate and reliable. The invention further discloses a virus genotyping detection kit with the probe assembly.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a probe combination, kit and method for virus genotyping detection. Background technique [0002] Cervical cancer is one of the most common malignant tumors of the female reproductive tract. Studies have shown that persistent infection and multiple infections of high-risk human papillomavirus (HPV) are one of the important causes of cervical cancer. Global research The results showed that the presence of high-risk HPV DNA was detected in 99.7% of cervical cancer patients. There are more than 200 types of HPV that have been discovered so far, mainly including low-risk types and high-risk types. The correlation between different types and different diseases is also different. Low-risk HPV infection may cause genital warts, while high-risk HPV infection is related to cervical cancer and vaginal cancer. At present, there is still a lack of effective treatments for HPV, so ea...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6834C12Q1/708C12Q2563/131C12Q2565/50C12Q2563/125
Inventor 廖玮莫亚勤林晓燕张晨光
Owner GUANGZHOU EZLIFE BIO CO LTD