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
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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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