Human herpesvirus I/II/III/V type nucleic acid typing detection kit and detection method

A detection kit and herpes virus technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of low detection sensitivity, long detection time, cumbersome operation, etc., and achieve high sensitivity Good, high specificity, effect before widespread use

Pending Publication Date: 2019-10-11
SHANGHAI BIOGERM MEDICAL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] (3) 12 Encephalitis Virus Nucleic Acid Multiplex PCR Detection Kits and Their Application (CN201510886216.7), including the detection of human herpesvirus I/II/III/V, the detection method is capillary electrophoresis, and the operation is relatively cumbersome and difficult to detect The time is long, the detection sensitivity is lower than that of fluorescent quantitative PCR method, and it is easy to produce false positives
[0010] Howe...

Method used

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  • Human herpesvirus I/II/III/V type nucleic acid typing detection kit and detection method
  • Human herpesvirus I/II/III/V type nucleic acid typing detection kit and detection method
  • Human herpesvirus I/II/III/V type nucleic acid typing detection kit and detection method

Examples

Experimental program
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Embodiment 1

[0079] Example 1——A human herpesvirus I / II / III / V type nucleic acid typing detection kit

[0080] A kit for detecting human herpesvirus type I / II / III / V nucleic acid typing by real-time fluorescent quantitative PCR, including: virus nucleic acid rapid extraction reagent, PCR amplification reagent, human herpesvirus type I / II / III / V Nucleic acid detection reagents, positive control substances, negative control substances. in:

[0081] (1) The viral nucleic acid rapid extraction reagent includes a lysis reagent, specifically including: Tris-HCl (pH10.0), Tween20, guanidine isothiocyanate, KCl, 4mM EDTA (pH8.0), and the reagent preparation method is shown in Table 4 Shown:

[0082] Table 4: Reagent Recipe for Rapid Viral Nucleic Acid Extraction

[0083]

[0084]

[0085] (2) The PCR amplification reagents include PCRbuffer, MgCl2, Tris-HCl (pH10.0), dATP, dCTP, dGTP, dTTP, DNA polymerase, BSA, and the reagent preparation method is as shown in Table 5:

[0086] Table 5: PCR...

Embodiment 2

[0110] Embodiment 2——A kind of detection method of influenza A virus H8N7

[0111] A real-time fluorescent PCR multiple detection method for human herpesvirus type I / II / III / V comprises the following experimental steps:

[0112] (1) Main reagents and instruments: the kit reagents in Example 1 were used; the fluorescent quantitative PCR instrument was ABI7500.

[0113] (2) Specimen preparation: Positive samples are inactivated virus after titration of human herpesvirus type Ⅰ, inactivated virus of human herpesvirus type II after titration, inactivated virus of human herpesvirus type III after titration, human herpesvirus type V After titration, the virus was inactivated, and then diluted with DEPC water in different multiples. The negative control specimen was a saliva swab of a healthy person.

[0114] (3) RNA extraction: Mix equal volumes of the viral nucleic acid rapid extraction reagent and the sample to be tested, let stand at room temperature for 5-10 minutes, and then di...

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Abstract

The invention discloses a human herpesvirus I/II/III/V type nucleic acid typing detection kit. The kit comprises a rapid virus nucleic acid extraction reagent, a PCR amplification reagent, a human herpesvirus I/II/III/V type nucleic acid detection reagent, a positive reference substance and a negative reference substance. According to the kit, multiple channel primer probes with human herpesvirusI/II/III/V type specificity are introduced so that the workload can be decreased, the problem is also avoided that other detection methods are not high in specificity and easily cause missed diagnosisand misdiagnosis, and it can be achieved that the same system can multiply and simultaneously detect four pathogens without mutual interference of the primer probes of all channels. The invention notonly discloses a real-time fluorescence quantitative PCR method aiming at human herpesvirus I/II/III/V type typing detection for the first time, but also has the advantages of good sensitivity, highspecificity, accuracy, reliability, rapidness, convenience and the like, thereby realizing rapid diagnosis of the infection situation of a herpesvirus I/II/III/V type. The invention also provides a human herpesvirus I/II/III/V type nucleic acid typing detection method.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnostic reagents, in particular to a human herpesvirus type I / II / III / V nucleic acid typing detection kit and detection method. Background technique [0002] Herpesviruses are a class of enveloped DNA viruses that primarily invade tissues of ectodermal origin, including skin, mucous membranes, and nervous tissue. The site of infection and the diseases it causes are various, and there is a tendency of latent infection, which seriously threatens human health. Currently known human herpes viruses include eight types, namely, herpes simplex virus type 1 (human herpes virus type I), herpes simplex virus type 2 (human herpes virus type II), varicella-zoster virus (human herpes virus type III) Type), Epstein-Barr virus (human herpesvirus type IV), cytomegalovirus (human herpesvirus type V), human herpesvirus type VI, human herpesvirus type VII, human herpesvirus type VIII. [0003] Herpes simplex (h...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/705C12Q1/686C12Q2563/107C12Q2545/114
Inventor 蒋小琴朱兆奎赵百慧
Owner SHANGHAI BIOGERM MEDICAL TECH CO LTD
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