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Gene chip for simultaneous detection of human herpesvirus and enterovirus

A technology of human herpes virus and enterovirus, applied in the field of molecular biology, can solve the problems of unsuitable virus extraction, cumbersome operation, virus loss, etc., and achieve the effect of fast extraction, high amplification efficiency and small sample size

Inactive Publication Date: 2016-04-06
山东大学齐鲁儿童医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Although the above literature discloses a method for simultaneously extracting DNA and RNA, it contains many reagents, complex components, and complicated operations. More importantly, this method is only aimed at RNA virus and DNA virus nucleic acid in animal serum and double swab samples. Because these samples usually contain high concentrations of RNA virus and DNA virus nucleic acid, it is easy to extract and separate, but if it is applied to cerebrospinal fluid samples with low virus content, it may cause the low concentration of virus in the cerebrospinal fluid itself to be lost and the result will be false negative, so it is not suitable Virus extraction from cerebrospinal fluid

Method used

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  • Gene chip for simultaneous detection of human herpesvirus and enterovirus
  • Gene chip for simultaneous detection of human herpesvirus and enterovirus
  • Gene chip for simultaneous detection of human herpesvirus and enterovirus

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Embodiment 1

[0074] A gene chip for simultaneously detecting human herpes virus and enterovirus in cerebrospinal fluid, including a nucleotide extraction kit, a DNA and RNA simultaneous reverse transcription kit, a DNA and RNA simultaneous PCR amplification kit, and a specific probe connected gene chip;

[0075] The specific reverse transcription primer sequences in the DNA and RNA simultaneous reverse transcription kit are as follows:

[0076] EV-r: 5'-TGACTCATCGACCTGATCTACA-3';

[0077] The primers in the described DNA and RNA simultaneous PCR amplification kit are composed of Biotin-HHV1,2,4,5-f, HHV1,2,5-r, HHV4-r, Biotin-EV-f and EV-r5 The primers are mixed in a molar ratio of 4.5:0.6:0.2:4.5:0.6, and the nucleotide sequences are as follows:

[0078] HHV1,2,4,5-f:5′-TCATCTACGGGGACACGGAC-3′

[0079] HHV1,2,5-r:5′-CGCACCAGATCCACGCCCTT-3′

[0080] HHV4-r:5′-GAGCTCCACCCCCTTCATC-3′

[0081] EV-f:5′-TTAAAACAGCCTGTGGGTTG-3′

[0082] EV-r:5′-TGACTCATCGACCTGATCTACA-3

[0083] Wherein HH...

Embodiment 2

[0121] A gene chip for simultaneously detecting human herpes virus and enterovirus in cerebrospinal fluid, including a nucleotide extraction kit, a DNA and RNA simultaneous reverse transcription kit, a DNA and RNA simultaneous PCR amplification kit, and a specific probe connected Gene chip; It is characterized in that:

[0122] The specific reverse transcription primer sequences in the DNA and RNA simultaneous reverse transcription kit are as follows:

[0123] EV-r: 5'-TGACTCATCGACCTGATCTACA-3';

[0124] The primers in the described DNA and RNA simultaneous PCR amplification kit are composed of Biotin-HHV1,2,4,5-f, HHV1,2,5-r, HHV4-r, Biotin-EV-f and EV-r5 The primers are mixed in a molar ratio of 4.5:0.6:0.2:4.5:0.6, and the nucleotide sequences are as follows:

[0125] HHV1,2,4,5-f:5′-TCATCTACGGGGACACGGAC-3′

[0126] HHV1,2,5-r:5′-CGCACCAGATCCACGCCCTT-3′

[0127] HHV4-r:5′-GAGCTCCACCCCCTTCATC-3′

[0128] EV-f:5′-TTAAAACAGCCTGTGGGTTG-3′

[0129] EV-r:5′-TGACTCATCGACCTGA...

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Abstract

The invention relates to a gene chip capable of simultaneously detecting human herpes virus and enterovirus. The gene chip comprises a nucleotide extraction kit, a simultaneous DNA (Deoxyribonucleic Acid) and RNA (Ribonucleic Acid) reverse transcription kit, a simultaneous DNA and RNA PCR amplification kit and a gene chip linked with a specific probe. According to the invention, the simultaneous extraction, amplification and detection of nucleic acid of a plurality of herpes viruses and intestinal viruses can be realized by only 100 mu l of cerebrospinal fluid. The gene chip has the advantages of less required sample amount, fast extraction speed, high amplification efficiency, reliable results and strong specificity. The early diagnosis and treatment of clinical viral encephalitis and meningitis induced by the viruses can be achieved and the current bottleneck problem of the detection of viruses in clinical cerebrospinal fluid is resolved.

Description

technical field [0001] The invention relates to a gene chip for simultaneously detecting human herpes virus and enterovirus, and belongs to the technical field of molecular biology. Background technique [0002] Viral encephalitis and meningitis are infectious diseases caused by viral infection and mainly damage the central nervous system (CNS). Common viruses mainly include herpes simplex virus type 1 and type 2 (HSV1 and HSV2), Varicella-zoster virus (VZV), Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), human herpesvirus types 6 (HHV6) and 7 (HHV7), and enteroviruses (EVs) that cause HFMD Such as Enterovirus 71 (EV71), Coxsackievirus A16 (CoxA16) and Coxsackie Group B 5 (CoxB5). Herpes simplex virus encephalitis caused by herpes simplex virus is the most common pathogen of sporadic encephalitis and meningitis. If acyclovir is not used for antiviral treatment in time, the mortality rate can be as high as 70%. In recent years, the scale of hand, foot and mouth dis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6837C12Q1/686C12Q1/70C12Q2531/113C12Q2565/501
Inventor 刘毅段春红马衍辉
Owner 山东大学齐鲁儿童医院
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