Internal-reference-containing HCMV fluorescence quantitative PCR detection kit

A detection kit and fluorescence quantitative technology, applied in the field of molecular biology, can solve problems such as difficult promotion, pollution, operation steps and long detection time, and achieve the effect of monitoring effectiveness and reducing false negative rate

Inactive Publication Date: 2011-07-06
SHANGHAI XINGYAO MED TECH DEV CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at the same time, the operation steps and detection time are long, and the experiment is easy to cause pollution, so it is difficult to carry out extensive clinical promotion

Method used

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  • Internal-reference-containing HCMV fluorescence quantitative PCR detection kit
  • Internal-reference-containing HCMV fluorescence quantitative PCR detection kit

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Experimental program
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Embodiment Construction

[0046] Design and prepare primers and probe sequences (for HCMV standard strain-Human herpesvirus 5 strain AD169 gene Sequence design):

[0047] Primer 1: 5`-GACTTCTTCACCCTGTTCTTCC-3` (Seq No.1)

[0048] Primer 2: 5`-GAGCCCGACTTTACCATCCA-3` (Seq No. 2)

[0049] Probe 1: 5`-CTATCAGAGATCACGATACAGCCGG-3` (Seq No.3)

[0050] Probe 2: 5`-CACTCAACCTACACGTCAACGCTA-3` (Seq No.4)

[0051] The above-mentioned primers and probes were synthesized in Bao Bioengineering (Dalian) Co., Ltd.

[0052] Internal reference male model: 5`-GACTTCTTCA CCCTGTTCTT CCTCGCACTC AACCTACACG TCAACGCTAC GGTATCGATA

[0053] ATCTTGTTGC GGTACTGGAT GGTAAAGTCG GGCTC-3`(Seq No.5)

[0054] Calibrator: 5`-GACTTCTTCA CCCTGTTCTT CCTCGCTATC AGAGATCACG ATACAGCCGG CGGTATCGAT

[0055] AATCTTGTTG CGGTACTGGA TGGTAAAGTC GGGCTC-3` (Seq No. 6)

[0056] The composition of the HCMV fluorescent quantitative PCR detection kit with internal reference is (20 people):

[0057] Composition name Volume

[0058] Nucleic Acid Extraction Solution 1ml

[00...

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Abstract

The invention relates to an internal-reference-containing human cytomegalovirus (HCMV) fluorescence quantitative polymerase chain reaction (PCR) detection kit. A detection primer and a detection probe are designed for a UL122 gene of an HCMV genome, a competitive internal reference amplification system is introduced by designing an internal reference probe and an internal reference male die and exists in the same reaction tube with an HCMV deoxyribonucleic acid (DNA) detection system, and the probability of false negative caused by the failure of the kit or experiment misoperation and the like can be effectively reduced by introducing the internal reference amplification system. The kit can perform qualitative and quantitative detection on HCMV DNA quickly and simply, and has high specificity and sensitivity.

Description

Technical field [0001] The invention relates to a HCMV fluorescent quantitative PCR detection kit containing internal reference, which detects both internal reference DNA and HCMV DNA, which can effectively prevent false negatives during the real-time fluorescent PCR detection of human cytomegalovirus, and belongs to molecular biology Field of study. Background technique [0002] Human cytomegalovirus (HCMV), also known as Humanherpesvirus 5 (Humanherpesvirus 5), belongs to the subfamily a herpesvirus. HCMV is one of the most common pathogens of human ancestral virus infection, which can infect the central nervous system of children System involvement, multiple organ damage, mental decline, deafness, etc.; it can cause mononucleosis in normal healthy people; it is like organ transplant recipients in immunodeficiency patients, acquired immunodeficiency syndrome patients, and cancer patients It can cause serious infections and even death. Together with toxoplasma, rubella virus, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 何剑军夏懿吴大治韩倩沈维祥
Owner SHANGHAI XINGYAO MED TECH DEV CO LTD
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