Unlock instant, AI-driven research and patent intelligence for your innovation.

Primer, probe and kit for fluorescence quantitative PCR (polymerase chain reaction) detection and parting of salmonella

A fluorescence quantitative and Salmonella technology, which is applied in the determination/inspection of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve the problems of detection and differentiation without diagnostic methods, and achieve the goal of lowering requirements and extensive diagnosis Effect

Inactive Publication Date: 2014-09-24
YANGZHOU UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no single diagnostic method can easily detect and differentiate all strains of Salmonella (two species of Salmonella, six subspecies of Salmonella enterica)

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
  • Primer, probe and kit for fluorescence quantitative PCR (polymerase chain reaction) detection and parting of salmonella
  • Primer, probe and kit for fluorescence quantitative PCR (polymerase chain reaction) detection and parting of salmonella
  • Primer, probe and kit for fluorescence quantitative PCR (polymerase chain reaction) detection and parting of salmonella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: PCR amplification of Salmonella standard strain

[0048] Fifteen strains of preserved Salmonella enterica subspecies typhi serotype were revived with blood plates and cultured. Bacterial colonies were placed in 1XPBS buffer, placed in a water bath at 100°C for 5 minutes, centrifuged at 10,000 rpm / min at 4°C for 10 minutes, and 10 μl was taken as a PCR template. The technique of the present invention is used for PCR amplification and high-resolution melting curve analysis (the specific technical details are as described above). The results showed that PCR can specifically and efficiently amplify Salmonella enterica subspecies enterica typhi serotype, and a melting temperature of 69°C appeared.

Embodiment 2

[0049] Example 2: Detection of Salmonella in healthy dog ​​feces

[0050] The collected fresh feces were immediately frozen and stored in a -80°C refrigerator, and then 200 mg was taken out for nucleic acid extraction (use Kit method), take 10 μl as a PCR template. The technique of the present invention is used for PCR amplification and high-resolution melting curve analysis (specific technical details and parameters are as described above). Of the 127 stool samples tested, 10 were positive (10 / 127, 7.9%). Melting curve analysis showed and confirmed by gene sequencing that 7 cases were Salmonella enterica subsp. enterica (melting temperature 69°C), 2 cases were Salmonella enterica subsp. Salmonella subsp. Arizona (melting temperature 54.5°C).

[0051]

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 a primer, a probe and a kit for fluorescence quantitative PCR (polymerase chain reaction) detection and parting of salmonella. The primer sequence is as shown in SEQ ID NO.1 and 2; the probe is as shown in SEQ ID NO.3 and 4; the bacteria to be tested are subjected to PCR augmentation by the primer and the probe; a 210bp strip is augmented; a salmonella positive sample enhanced at 640nm of wavelength is detected by fluorescence; and then the salmonella can be parted according to the melting temperature. The primer is significant in technical advantages, and has high specificity and sensitivity; and the main pathogenic bacteria (intestinal salmonella subspecies) and other subspecies of Bongor salmonella and intestinal salmonella can be conveniently parted. Compared with the technology disclosed by the invention, the primer can be conveniently and widely used for diagnosing infection of human and animal salmonella and bacterial food poisoning.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR detection reagent and a detection method for Salmonella molecular diagnosis. This invention effectively amplifies all strains of Salmonella (two species of Salmonella and six subspecies of Salmonella enterica). Furthermore, the high-resolution melting curve technique conveniently classified Salmonella into 3 groups: i) the important pathogenic enteric subspecies (melting temperature 69.5°C), ii) Howden subsp. La subsp. (63.8°C), iii) Salmonella Bongor (54.8°C), Indiga subsp. (57°C), Salam subsp. (55.5°C), Arizona subsp. (54.5°C). Background technique [0002] Salmonella spp. can cause various clinical manifestations of salmonellosis in humans and animals, and is also one of the main causes of human food poisoning, which is very important to medicine, veterinary medicine and public health. The genus Salmonella is divided into S. enterica and S. bongori. Salmonella enterica is divided into 6 subs...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/10C12Q1/06C12N15/11
CPCY02A50/30
Inventor 王成明徐步王志强许传灵危蓝菁张继垒
Owner YANGZHOU UNIV