Barcode identification primer, identification method and kit for different species in aeromonas

An Aeromonas, identification method technology, applied in the field of biology, can solve problems such as inability to accurately identify species, and achieve the effect of simple operation, high precision and accuracy

Active Publication Date: 2020-11-13
UNIV OF ELECTRONICS SCI & TECH OF CHINA ZHONGSHAN INST +1
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The difference between the 16SrRNA genes of different species of Aeromonas is low, only 0.06% to 0.66%. According to the commonly used standard, if the sequence similarity reaches more than 97%, it is regarded as the same level. However, using 16SrRNA sequence for analysis may cause The similarity with two or more species is greater than 97%, which leads to the inability to accurately identify the species

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Barcode identification primer, identification method and kit for different species in aeromonas
  • Barcode identification primer, identification method and kit for different species in aeromonas
  • Barcode identification primer, identification method and kit for different species in aeromonas

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1.1 Design and synthesis of primers

[0023] Using the cpn60 gene sequence of Aeromonas victorii in Genbank (accession number EU306841.1) as the target gene, the primers were designed using DNAStar-Lasergene software. The primers were synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd. Table 1.

[0024] Table 1 The specific primer sequence of cpn60 gene

[0025]

[0026] 1.2 Activation of strains and extraction of genome

[0027] 1.2.1 Method

[0028] (1) Activation of the strain: Take out the preserved glycerol bacteria from the ultra-low temperature refrigerator, inoculate 50 μl of the bacteria solution with a pipette gun in a sterile environment at a ratio of 1:200, and inoculate it into 10 ml of LB liquid medium, at 37°C , 140rpm shaking culture overnight.

[0029] (2) Bacteria collection: Take 1ml of the culture in a 1.5ml Ep tube, centrifuge at room temperature at 8000rpm for 5min, discard the supernatant, refill the culture into the Ep tube, repeat t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses primers, a method and a kit for identifying barcodes of different species in aeromonas. The sequences comprise F: 5 '-GGAATTCGCAACCGTTCTGGCTCAGGCTATCGT-3' and R: 5 '-TGTCGACGACGGCAGCCACTTTCAC-3'. The 16S rRNA sequence analysis and identification method generally considers that when the sequence similarity reaches 97% or above, it is regarded as the same level, and in practical application, multiple different species of aeromonas often meet the sequence similarity requirement. In this case, 16S rRNA sequence analysis can only identify the genus level but cannot accurately identify the species. According to the method, the problems can be solved, after BLAST comparison is conducted on the cpn60 sequence of the species to be detected, the comparison result with the similarity larger than 98.8% and the coverage larger than 98% is highly specific, the cpn60 sequence is a specific species under aeromonas, and different species do not exist in the comparison result.

Description

Technical field: [0001] The invention belongs to the technical field of biology, and in particular relates to a barcode identification primer, an identification method and a kit for different species in the genus Aeromonas. Background technique: [0002] Aeromonas spp belongs to Vibrioceae and is facultatively anaerobic. It is a Gram-negative bacterium widely distributed in aquatic habitats such as rivers, lakes, ponds, seawater, and estuaries. Aeromonas can infect a variety of animals, causing disease and even death, which poses a huge challenge to the aquaculture industry. In addition, pathogenic Aeromonas can also cause acute diarrhea, bacteremia, meningitis, endocarditis and other diseases in humans, bringing hidden dangers to public health safety and affecting human health. Therefore, how to quickly and accurately identify pathogenic bacteria, so as to control the disease in a targeted manner, is of great significance in production practice. [0003] At present, the d...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/689C12Q1/6869C12Q1/04C12N15/11
CPCC12Q1/689C12Q1/6869C12Q2531/113C12Q2565/125C12Q2537/165
Inventor李梅许承佳李丹林森馨陈发明李天宇杨呈勇刘贻尧吴国锋潘育方
OwnerUNIV OF ELECTRONICS SCI & TECH OF CHINA ZHONGSHAN INST