Primer probe composition and application thereof in preparation of parainfluenza virus typing detection kit

A technology for detection kits and primer probes, applied in the preparation of parainfluenza virus typing detection kits, in the field of primer probe compositions, can solve the problem of inability to effectively distinguish human parainfluenza virus types 1 and 4, specific issues of low sex

Pending Publication Date: 2021-09-14
首都儿科研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the primers and probes used in the above kits have excellent detection sensitivity for parainfluenza virus types 4a and 4b, their specificity is still low and cannot effectively distinguish human parainfluenza virus types 1 and 4

Method used

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  • Primer probe composition and application thereof in preparation of parainfluenza virus typing detection kit
  • Primer probe composition and application thereof in preparation of parainfluenza virus typing detection kit
  • Primer probe composition and application thereof in preparation of parainfluenza virus typing detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Specific primers and probes were designed for parainfluenza virus type 1 HN gene, parainfluenza virus type 2 L gene, parainfluenza virus type 3 L gene, parainfluenza virus type 4a L gene and parainfluenza virus type 4b N gene, and The synthesis was entrusted to Sangon Bioengineering (Shanghai) Co., Ltd.; the nucleotide sequence information of the four sets of primers and probes obtained is shown in Table 1.

[0034] Table 1 Primer and probe composition sequence information

[0035]

[0036]Among them, the length of the HN gene amplification product of parainfluenza virus type 1 is 90 bp, the length of the amplification product of parainfluenza virus type 2 L gene is 137 bp, the length of the amplification product of parainfluenza virus type 3 L gene is 128 bp, and the length of the amplification product of parainfluenza virus type 4a L gene is 128 bp. The length of the gene amplification product is 90bp, and the length of the parainfluenza virus type 4b N gene amplif...

Embodiment 2

[0058] After the determination of concentration and purity, each positive gene containing parainfluenza virus type 1 HN gene, parainfluenza virus type 2 L gene, parainfluenza virus type 3 L gene, parainfluenza virus type 4a L gene and parainfluenza virus type 4b N gene The plasmids were serially diluted 10 times to get from 2.4×10 1 ~2.4×10 6 A total of 6 dilutions of positive plasmids in copies / μL were used as standard templates. Prepare the PCR reaction system according to Table 5 in Example 1, carry out multiple RT-qPCR reactions according to the reaction program shown in Table 6, and obtain the fluorescence amplification curve (such as figure 1 , figure 2 , image 3 , Figure 4 and Figure 5 shown), draw a standard curve and determine the lowest detection limit. Depend on figure 1 , figure 2 , image 3 , Figure 4 and Figure 5 shown at a concentration of 1.2 x 10 2 ~1.2×10 8 Within the range of copies / μL, the kit of the present invention amplifies the fluor...

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Abstract

The invention discloses a primer probe composition and application thereof in preparation of a parainfluenza virus typing detection kit. The primer probe composition at least comprises a primer and a probe for detecting parainfluenza virus 4a, and a primer and a probe for detecting parainfluenza virus 4b. The primer probe composition can be used for specifically typing parainfluenza virus 4a and parainfluenza virus 4b, and has high detection sensitivity. The detection proves that the lowest detection limit of the primer probe composition for parainfluenza virus 4a is 4.8 * 10<1> copies/ microliter, and the lowest detection limit for parainfluenza virus 4b is 5.2 * 10<1> copies/ microliter, so the primer probe composition has high sensitivity. The primer probe composition can be used for specifically typing parainfluenza virus 4a and parainfluenza virus 4b, does not have cross reaction with parainfluenza virus 1, 2 or 3, has very high specificity, and can quickly and accurately perform typing detection on parainfluenza virus.

Description

technical field [0001] The invention belongs to the technical field of virus detection, in particular to a primer probe composition and its application in preparing a parainfluenza virus typing detection kit. Background technique [0002] Parainfluenza virus is a kind of enveloped single-stranded negative-strand RNA virus belonging to the genus Paramyxovirus. It is a common pathogen of respiratory tract infection. It mainly causes severe lower respiratory tract infection in infants and children. At present, there is no effective treatment. Drugs and virus vaccines. [0003] According to heredity and antigenicity, parainfluenza virus can be divided into type 1, type 2, type 3 and type 4. Type 1-3 parainfluenza virus is an important pathogen that causes acute respiratory infection in infants and young children, and type 4 only Causes respiratory tract infection in infants with mild symptoms, and type 4 can be further divided into type 4a and type 4b. The symptoms of infectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12Q1/6848C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q1/6848C12Q2600/16C12Q2531/113C12Q2521/107C12Q2563/107C12Q2537/143C12Q2521/531
Inventor 赵林清张迪骏陈冬梅
Owner 首都儿科研究所
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