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Primers, probe and method for detecting various respiratory viruses

A respiratory and probe technology, applied in the field of virus diagnosis, can solve the problems of cumbersome procedures and inability to diagnose coronavirus, etc., achieve broad application prospects, save resources, and reduce operating steps

Inactive Publication Date: 2012-05-02
SHANGHAI PUBLIC HEALTH CLINICAL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the invention has a large throughput and speeds up the detection to a certain extent, this method first needs to couple the nucleic acid probe to the microsphere, and finally needs to carry out the PCR amplification product of the sample with the nucleic acid probe microsphere group. The procedure of hybridization and labeling is relatively cumbersome. In addition, this method cannot diagnose viruses such as coronavirus OC43 and 229E / NL63. Therefore, there is an urgent need for a faster, economical and effective detection method that can detect multiple respiratory viruses at the same time.

Method used

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  • Primers, probe and method for detecting various respiratory viruses
  • Primers, probe and method for detecting various respiratory viruses
  • Primers, probe and method for detecting various respiratory viruses

Examples

Experimental program
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Effect test

Embodiment 1

[0053] Detection of multiple respiratory viruses using the primers and probes of the invention

[0054] 1 Materials and methods

[0055] 1.1 Sample source

[0056] From June 7, 2009 to June 5, 2010, 0-1-year-old children with sporadic upper respiratory tract infection symptoms such as fever, cough, runny nose and sore throat who visited the fever clinic of Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were collected through cooperative relations. 62 cases of children were tested, and 9 cases were selected, which were recorded as sample 1 ~ sample 9 respectively.

[0057] 1.2 Sample collection, extraction and reverse transcription of viral nucleic acid

[0058] Use a throat swab to collect mucus from the upper respiratory tract of the child. Throat swabs were placed in 4ml of transport solution (containing Hanks solution and 10% glycerol), transported to the laboratory at 4°C, and stored at -70°C for later use. The viral RNA / DNA extraction...

Embodiment 2

[0105] Embodiment 2 Kit 1 of the present invention

[0106] The kit contains the following reagents and materials:

[0107] 1. Primer solution (100 μmol / L): composed of primer pairs SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO. 9-10

[0108] 2. Probe solution (100μmol / L): composed of probe SEQ ID NO.31~35

[0109] 3. dNTPs (2.5mol / L)

[0110] 4. 10×PCR Buffer (Mg 2+ Plus)

[0111] 5. rTaq enzyme (5U / μl)

Embodiment 3

[0112] Embodiment 3 Kit 2 of the present invention

[0113] The kit contains the following reagents and materials:

[0114] 1. Primer solution (100 μmol / L): composed of primer pairs SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO. 19-20

[0115] 2. Probe solution (100μmol / L): composed of probe SEQ ID NO.36~40

[0116] 3. dNTPs (2.5mol / L)

[0117] 4. 10×PCR Buffer (Mg 2+ Plus)

[0118] 5. rTaq enzyme (5U / μl)

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Abstract

The invention provides a primer combination for detecting various respiratory viruses, a specific probe combination for detecting various respiratory viruses, a method using the primers and probe to detect various respiratory viruses and a kit comprising the primer combination and the probe combination. In the invention, the mixed solution of the specific primers and the probe with fluorescent mark is utilized to carry out PCR (Polymerase Chain Reaction) amplification on batch samples, and then microsatellite analysis is performed, so as to analyze a great quantity of samples at one time and detect various respiratory viruses simultaneously, thus significantly improving efficiency and realizing high sensitivity. The kit can be popularized and used clinically, and has wide application prospect.

Description

technical field [0001] The invention relates to the technical field of virus diagnosis, in particular to a primer, a probe and a method for detecting various respiratory viruses. Background technique [0002] Respiratory virus is a kind of virus that can invade the respiratory tract and cause local lesions of the respiratory tract or only use the respiratory tract as the entry point to cause lesions of tissues and organs outside the respiratory tract. According to statistics, more than 90% of acute respiratory infections are caused by viruses, which are highly contagious and pathogenic, and can be directly transmitted from person to person through saliva droplets. Therefore, the diagnosis of respiratory viruses must be strengthened to actively treat and prevent transmission. [0003] Respiratory viruses include influenza viruses in the family Orthomyxoviridae; parainfluenza viruses in the family Paramyxoviridae, respiratory syncytial virus, measles virus, mumps virus, and s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 周晓明胡芸文张璐王宏萍张国翠
Owner SHANGHAI PUBLIC HEALTH CLINICAL CENT
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