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Kit and method for simultaneously detecting herpes simplex virus, Carbosarcoma associated herpes virus, JC virus and EB virus

A herpes simplex virus, herpes virus technology, applied in the field of real-time fluorescent PCR detection kits, can solve the problem of inability to detect herpes simplex virus Kaposi's sarcoma-related herpes virus, JC virus and EB virus at the same time

Active Publication Date: 2021-01-08
浙江安维珞诊断技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem that herpes simplex virus, Kaposi's sarcoma-associated herpesvirus, JC virus and Epstein-Barr virus cannot be detected simultaneously in the prior art, the present invention provides herpes simplex virus, Kaposi's sarcoma-associated herpesvirus, Detection primers and probes for JC virus and EB virus nucleic acids to achieve rapid, efficient and accurate detection of these four viruses

Method used

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  • Kit and method for simultaneously detecting herpes simplex virus, Carbosarcoma associated herpes virus, JC virus and EB virus
  • Kit and method for simultaneously detecting herpes simplex virus, Carbosarcoma associated herpes virus, JC virus and EB virus
  • Kit and method for simultaneously detecting herpes simplex virus, Carbosarcoma associated herpes virus, JC virus and EB virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] [Example 1] A kit for detecting herpes simplex virus, Kaposi's sarcoma-associated herpes virus, JC virus and Epstein-Barr virus in the same reaction is provided.

[0069] The kit includes PCR reaction solution, nucleic acid composition for simultaneous detection of herpes simplex virus, Kaposi's sarcoma-associated herpes virus, JC virus and Epstein-Barr virus, hot start Taq enzyme, positive control and negative control. Wherein, the PCR reaction solution includes 250mM Tris-base, 0.25% TritonX-100, 25mmol / L MgCl 2 .

[0070] Wherein the nucleic acid composition comprises:

[0071] A pair of herpes simplex virus amplification primers whose sequences are shown in SEQ ID No.1 and SEQ ID No.2;

[0072] Kaposi's sarcoma-associated herpesvirus amplification primer pair whose sequences are shown in SEQ ID No.4 and SEQ ID No.5;

[0073] The JC virus amplification primer pair whose sequences are shown in SEQ ID No.7 and SEQ ID No.8;

[0074] The Epstein-Barr virus amplificat...

Embodiment 2

[0082] [Example 2] Determination of the sensitivity of the kit in Example 1 to detect four viruses in the same reaction

[0083] (1) Herpes simplex virus positive control substance, Kaposi's sarcoma-associated herpesvirus positive control substance, JC virus positive control substance, and EB virus positive control substance in the kit of Example 1 are mixed and made into a mixed solution to obtain positive to be treated Test product (the concentration of each virus positive control substance is 10 6 copies / mL).

[0084] (2) Perform a 10-fold serial dilution of the positive sample to be tested (i.e. 10copies / mL~10 5 copies / mL), using the kit of Example 1 to perform multiple fluorescent quantitative PCR detection on the positive test items under each gradient, the system of multiple fluorescent quantitative PCR reactions is shown in Table 1. The reaction conditions of the multiplex fluorescent quantitative PCR reaction were: pre-denaturation at 95°C for 3 min; denaturation at...

Embodiment 3

[0090] [Example 3] The specificity comparison between the kit of Example 1 and known commercialized kits

[0091] (1) Herpes simplex virus positive control substance, Kaposi's sarcoma-associated herpesvirus positive control substance, JC virus positive control substance, EB virus positive control substance in the test kit of embodiment 1 (the concentration of each virus positive control substance is equal to for 10 6 copies / mL) were mixed and made into a mixed solution to obtain a positive test article; physiological saline not containing the above four virus positive control substances was a negative test article.

[0092] (2) The experiment is divided into an experimental group, a control group 1 and a control group 2, and the kit of the experimental group is the kit of Example 1. The test kit of matched group 1 is roughly the same as the kit of embodiment 1, and the kit of matched group 2 is roughly the same as the kit of embodiment 1 (the kit components of matched group 1...

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Abstract

The invention discloses a kit and method for simultaneously detecting herpes simplex virus, Carbosarcoma-associated herpes virus, JC virus and EB virus. According to the kit and the method, a real-time fluorescent quantitative PCR technology is applied, highly specific primers and probes for herpes simplex virus, Carbosarcoma-associated herpes virus, JC virus and EB virus are adopted, and whetherthe four viruses exist in a sample to be detected or not can be judged through one PCR reaction, so that the method is more convenient and quicker than a single fluorescent quantitative PCR method, and the cost is saved. The detection sensitivity of the kit to four viruses reaches 10 copies / mL, and the kit has good specificity and does not have cross reaction with other viruses; and meanwhile, thekit is good in amplification repeatability, and the precision is smaller than or equal to 2%.

Description

technical field [0001] The invention belongs to the technical field of virus detection kits, and relates to a real-time fluorescent PCR detection kit, in particular to a kit for simultaneously detecting herpes simplex virus, Kaposi's sarcoma-associated herpes virus, JC virus and Epstein-Barr virus and a detection method thereof. Background technique [0002] Herpes simplex virus (HSV) can be divided into type I and type II (HSV-I and HSV-II). Humans are the only natural host of HSV. Type I herpes simplex disease is mainly transmitted through close contact with the respiratory tract, skin and mucous membranes, and infects the skin and mucous membranes and organs above the waist, such as causing inflammation and herpes in the mucous membranes of the lips, nasal vestibule, conjunctiva, and throat. 99% of herpes in and around the mouth are caused by herpes type I infection. Clinically, it can be divided into primary and recurrent herpes simplex. Common cases such as skin herp...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/705C12Q1/708C12Q1/686C12Q2600/16C12Q2537/143C12Q2563/107
Inventor 古柏燕彭忠
Owner 浙江安维珞诊断技术有限公司
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