Primer composition for simultaneously detecting seven respiratory tract related viruses, and application of primer composition

A primer composition and respiratory technology, applied in the field of biomedicine, can solve the problems of human respiratory syncytial virus typing and human parainfluenza virus typing, so as to avoid false positive results, short detection cycle and strong detection specificity Effect

Pending Publication Date: 2021-12-21
NINGBO HEALTH GENE TECHNOLOGIES CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For example, the Chinese invention patent application with publication number CN111808997A discloses a composition, kit, use and method for detecting and typing 10 respiratory-related viruses. Influenza virus, respiratory adenovirus, human parainfluenza virus type 1, human parainfluenza virus type 2, human parainfluenza virus type 3, human parainfluenza virus type 4, influenza A virus, human respiratory syncytial virus and metapneumovirus These ten respiratory-associated viruses; however, cannot type both types A and B of human respiratory syncytial virus, and cannot type types 4a and 4b of human parainfluenza virus

Method used

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  • Primer composition for simultaneously detecting seven respiratory tract related viruses, and application of primer composition
  • Primer composition for simultaneously detecting seven respiratory tract related viruses, and application of primer composition
  • Primer composition for simultaneously detecting seven respiratory tract related viruses, and application of primer composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] For the human sub-fluvophilic virus 1 L gene, human sub-flu virus type 2 L gene, human sub-flu virus 3 N gene, human subfluenza virus 4A L-gene, human beefire virus 4B type L gene, human respiratory traction Cytovirus A type L gene, human respiratory syncytial viral B-type L gene, human RNA internal parametrin B2M gene and human RNA interference IC gene design specific primer, and enriminating Shenggong Biological Engineering (Shanghai) Co., Ltd. Synthesis; 9 The nucleotide sequence information of the group is shown in Table 1.

[0048] Table 1 Nucleotide sequence information of each primer pair

[0049]

[0050]

[0051] The respiratory viral type detection kit of the present invention was prepared using the primer composition shown in Table 1, which contains an RT-PCR reaction liquid, an primer composition, an RT-PCR enzyme, a positive control, and a negative control. Among them, the composition of the primer composition is shown in Table 2, and the reagent compositio...

Embodiment 2

[0069] Clinical collection of other common respiratory viral infections other than the seven viruses shown in Example 1 (see Table 7), all samples have been used in Ningbo Haierch Gene Technology Co., Ltd. More respiratory pathogens The case (Machinery Temporation 20183400518) was diagnosed, and the sample S8-S17 was detected (the detection method was identical to Example 1) using the respiratory associated viral classification kit of the present invention to verify the respiratory-related viral type of the present invention. Detect the specificity of the kit.

[0070] Table 7 Clinical respiratory virus infection positive sample information

[0071] Sample number Virus infection type S8 甲 甲 流 virus H1N1 S9 甲 甲 流 virus H3N2 S10 Balline flu virus S11 Adenovirus S12 Nasal virus S13 Meticulosis S14 Ordinary coronary virus S15 Boda virus S16 Pneumoniae S17 Mycoplasma pneumoniae

[0072] The detection result shows that the ...

Embodiment 3

[0074] The measurement concentration and purity include a human sub-flu virus 1 L gene, a human sub-flu virus type 1 L gene, human sub-flu virus 3 type N gene, human sub-flu virus 4A type L gene, human sub-flu virus 4B type L gene, human respiratory syncytial viral A-type L gene, the recombinant plasmid of human respiratory syncytial viral B genotype 2 times gradient dilution, obtained from 1 × 10 1 ~ 6.4 × 10 2 COPIES / μL total 7 dilution positive particles were used as standard templates; and then the above-mentioned yang-based RT-PCR reaction was performed using the same method in Example 1 using the same method in Example 1. Positive recombinant plasmid of each concentration Fluorescence signal value (see Table 8).

[0075] Table 8 Different concentration positive recombinant plasmid fluorescent signal values ​​(RFU) summary table

[0076]

[0077] Note: Due to the detection platform, when the fluorescent signal value is greater than 30000, the detection platform will not b...

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Abstract

The invention discloses a primer composition for simultaneously detecting seven respiratory tract related viruses, and application of the primer composition. The primer composition can simultaneously realize detection and typing of human parainfluenza viruses type 1, type 2, type 3, type 4a and type 4b, and the type A and the type B of human respiratory syncytial viruses, and provides a powerful basis for targeted treatment after virus infection; each primer pair has strong detection specificity for corresponding viruses, does not have specific amplification on other common respiratory tract related viruses (such as influenza A virus, influenza B virus, adenovirus, rhinovirus, metapneumovirus, coronavirus, bocavirus, chlamydia pneumoniae and mycoplasma pneumoniae), and does not have a false positive result; and through detection, the lowest detection limit of the primer composition disclosed by the invention to the respiratory tract related viruses can reach 1.0 * 10 < 1 > copies/mu L, and sensitivity is high.

Description

Technical field [0001] The present invention belongs to the field of biomedical technology, and more particularly to primer compositions of seven respiratory related viruses and their applications. Background technique [0002] The human respiratory syncytial virus (HRSV) and human sub-flu virus (HPIVS) are the main pathogens that cause children and high-risk adult underwent diseases, and there is currently a lack of effective treatment of drugs and virus vaccines. [0003] Among them, the subfluenza virus is a class of condensing single-stranded RNA viruses belonging to the sub-adhesive velocity, mainly causes serious lower respiratory infections in infants and children. According to genetic and antigenicity, the subfluid virus can be divided into type 1, type 2, 3 and 4, 1-3 sub-flu virus is a more common pathogens that cause infants and young children's acute respiratory infections, and only 4 Infant infant respiratory infections in symptoms, and 4 types can be further divided...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2600/166C12Q2521/107C12Q2537/143C12Q2521/531C12Q2563/107C12Q2545/113Y02A50/30
Inventor 张迪骏赵林清罗勇陈冬梅徐智孙宇梁宜朱汝南曾县平钱渊余丁吴勇
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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