Detection marker, primer probe pair, kit and detection method for herpesvirus hominis I type and II type

A kit and virus technology, applied in the field of herpes simplex virus type I and/or type II detection markers, can solve the problems of cumbersome operation, difficult to meet early diagnosis, and few applications, so as to improve detection sensitivity and nucleic acid similarity Low, less cross-reactive effects

Active Publication Date: 2019-03-19
中生方政生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus isolation and culture takes a long period of time, cumbersome operations, professional operators and harsh laboratory conditions are required, so it is rarely used in clinical rapid diagnosis
[0008] 2) There is still lag in immunological diagnosis, which cannot accurately reflect whether the current infection or pathogen is carried, and it is difficult to meet the requirements of early diagnosis
The high similarity between HSV-1 and HSV-2 makes it difficult to detect HSV types

Method used

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  • Detection marker, primer probe pair, kit and detection method for herpesvirus hominis I type and II type
  • Detection marker, primer probe pair, kit and detection method for herpesvirus hominis I type and II type
  • Detection marker, primer probe pair, kit and detection method for herpesvirus hominis I type and II type

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1: be used for the design of the primer probe pair of rapid detection HSV-1 / HSV-2

[0082] According to the HSV-1RL2 gene, HSV2RS1 gene, and HBB gene sequences in NCBI, primer pairs and probes were designed. The sequences are as follows:

[0083] Table 1 Primer and Probe Sequences

[0084]

Embodiment 2

[0085] Embodiment 2: be used for the real-time fluorescent quantitative PCR kit of rapid detection HSV-1

[0086] Real-time fluorescent quantitative PCR kit for rapid detection of human HSV-1, including HSV-1 primers, HSV-1 probes, HBB primers, HBB probes, enzyme mixture, reaction buffer, sample extract, positive control , negative control substance, instructions and box body.

[0087] The sequences of primers HSV-1-F and HSV-1-R are as shown in SEQ ID NO.1-2 in sequence, and the concentration is 500nM; the sequence of probe HSV1-P is as shown in SEQ ID NO:3, and the fluorescent group is Cy5. The quenching group was BHQ3 at a concentration of 200 nM.

[0088] The sequences of the primers HBB-F and HBB-R are shown in SEQ ID NO.7-8 in turn, and the concentration is 500nM; the sequence of the probe HBB-P is shown in SEQ ID NO:3, and the fluorescent group of the probe is ROX, which is quenched The group is BHQ1 and the concentration is 200 nM.

[0089] Among them, Anstart qPCR ...

Embodiment 3

[0097] Embodiment 3: The real-time fluorescence quantitative PCR kit for rapid detection of HSV-2

[0098] Real-time fluorescent quantitative PCR kit for rapid detection of human HSV-2, including HSV-2 primers, HSV-2 probes, HBB primers, HBB probes, enzyme mixture, reaction buffer, sample extract, positive control , negative control substance, instructions and box body.

[0099] Wherein the sequences of primers HSV-2-F and HSV-2-R are shown in SEQ ID NO.4-5 in turn, and the concentration is 500nM; the sequence of probe HSV-2-P is shown in SEQ ID NO:6, and the fluorophore is FOM, the quenching group is BHQ1, the concentration is 200nM.

[0100] The sequences of the primers HBB-F and HBB-R are shown in SEQ ID NO.7-8 in turn, and the concentration is 500nM; the sequence of the probe HBB-P is shown in SEQ ID NO:3, and the fluorescent group of the probe is ROX, which is quenched The group is BHQ1 and the concentration is 200 nM.

[0101] Among them, Anstart qPCR Master Mix Enzym...

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Abstract

The invention relates to the technical field of detection of pathogenic microorganisms, in particular to a detection marker, primer probe pair, kit and detection method for herpesvirus hominis I typeand II type. By taking reverse repeated sequences RS1 and RS2 in an HSV genome as a target area, the detecting sensitivity can be further improved as two copies are available in the genome on the onehand and cross reactions are unlikely to be caused as the inter-nucleic acid similarity is low on the other hand.

Description

technical field [0001] The invention relates to the technical field of detection of pathogenic microorganisms, in particular to a detection marker of herpes simplex virus type I and / or type II, a pair of primers and probes, a kit and a detection method. Background technique [0002] Herpes simplex is a viral skin disease caused by herpes simplex virus, which can cause various human diseases, such as gingivostomatitis, keratoconjunctivitis, encephalitis, reproductive system infection and neonatal child's infection. [0003] Herpes simplex virus (HSV) belongs to the subfamily Avirinae of the family Herpesviridae, and the size of the virus plasmid is about 180 nanometers. According to the difference in antigenicity, the virus is currently divided into type I (HSV-1) and type II (HSV-2). HSV-1 is mainly obtained from lip lesions, and can also be isolated from genital lesions, and HSV-2 can be isolated from genital lesions. Infection is due to human-to-human contact, and it is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/705C12Q2561/113C12Q2531/113C12Q2563/107Y02A50/30
Inventor 魏颖颖蔺皓邹国宝刘欣欣宋高尚沈江卫吴茜
Owner 中生方政生物技术股份有限公司
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