CAMP primer group for amplifying salmonella, test kit and application

A Salmonella and primer set technology is applied in the field of CAMP primer sets for amplifying Salmonella, and can solve the problems of high cost, complex detection method and the like

Pending Publication Date: 2020-07-14
ZHONGKE XINRAY SUZHOU BIOLOGICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It aims to solve the technical problems of complex detection methods and high cost in the prior art

Method used

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  • CAMP primer group for amplifying salmonella, test kit and application
  • CAMP primer group for amplifying salmonella, test kit and application
  • CAMP primer group for amplifying salmonella, test kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Application of Eva Green to Verify the Amplification Reaction to Salmonella 1-Sa

[0048] Similar to SYBR Green I, Eva Green is a dye with a green excitation wavelength that binds to the minor groove region of all dsDNA double helices, and its inhibition on nucleic acid amplification reactions such as PCR is much smaller than that of the latter. In the free state, EvaGreen emits weak fluorescence, but once bound to double-stranded DNA, the fluorescence is greatly enhanced. Therefore, the fluorescence signal intensity of Eva Green is related to the amount of double-stranded DNA, and the amount of double-stranded DNA present in the nucleic acid amplification system can be detected based on the fluorescence signal.

[0049] Reaction solution mix (25 μL)

[0050] 20mM Tris-HCl pH8.8

[0051] 10mM KCl

[0052] 10mM (NH 4 ) 2 SO 4

[0053] 14mM MgSO 4

[0054] 0.1% Triton X-100

[0055] 1M betaine

[0056] 1.25mM dNTPs

[0057] 8U Bst DNA polymerase (NEW...

Embodiment 2

[0069] Example 2 Application of Eva Green to Verify the Amplification Reaction to Salmonella 2-Sa

[0070] Similar to SYBR Green I, Eva Green is a dye with a green excitation wavelength that binds to the minor groove region of all dsDNA double helices, and its inhibition on nucleic acid amplification reactions such as PCR is much smaller than that of the latter. In the free state, EvaGreen emits weak fluorescence, but once bound to double-stranded DNA, the fluorescence is greatly enhanced. Therefore, the fluorescence signal intensity of EvaGreen is related to the quantity of double-stranded DNA, and the quantity of double-stranded DNA existing in the nucleic acid amplification system can be detected according to the fluorescent signal.

[0071] Reaction solution mix (25 μL)

[0072] 20mM Tris-HCl pH8.8

[0073] 10mM KCl

[0074] 10mM (NH 4 ) 2 SO 4

[0075] 14mM MgSO 4

[0076] 0.1% Triton X-100

[0077] 1M betaine

[0078] 1.25mM dNTPs

[0079] 8U Bst DNA polymera...

Embodiment 3

[0091] Example 3 Application of Eva Green to Verify the Amplification Reaction to Salmonella 3-Sa

[0092] Similar to SYBR Green I, EvaGreen is a dye with a green excitation wavelength that binds to the minor groove region of all dsDNA double helices, and its inhibition on PCR and other nucleic acid amplification reactions is much smaller than that of the latter. In the free state, EvaGreen emits weak fluorescence, but once bound to double-stranded DNA, the fluorescence is greatly enhanced. Therefore, the fluorescence signal intensity of EvaGreen is related to the quantity of double-stranded DNA, and the quantity of double-stranded DNA existing in the nucleic acid amplification system can be detected according to the fluorescent signal.

[0093] Reaction solution mix (25 μL)

[0094] 20mM Tris-HCl pH8.8

[0095] 10mM KCl

[0096] 10mM (NH 4 ) 2 SO 4

[0097] 14mM MgSO 4

[0098] 0.1% Triton X-100

[0099] 1M betaine

[0100] 1.25mM dNTPs

[0101] 8U Bst DNA polymer...

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Abstract

The invention discloses a CAMP primer group for amplifying salmonella and a test kit. The CAMP primer group comprises one or more than two selected from the group consisting of the following three primer groups: a primer group for amplifying a CFSA231 complete genome of the salmonella, a primer group for amplifying a CFSA664 complete genome of the salmonella, and a primer group for amplifying a CFSA1096 complete genome of the salmonella. The CAMP primer group is obtained by designing a CFSA231 genome, a CFSA664 genome and a CFSA1096 genome of three specific conserved regions of the salmonella,and the genome of each conserved region is composed of six primers, and comprises a group of sequence main primers (NF / NR), a group of outer primers (F2 / R2) and a group of inner primers (LF / LR). Theprimer group provided by the invention is high in sensitivity and strong in specificity; and the test kit prepared from the primer group can rapidly and accurately detect the salmonella contained in ato-be-detected sample. The visual test kit provided by the invention provides great convenience for on-site detection, and can realize rapid and accurate detection of the salmonella by being preparedinto the test kit.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a CAMP primer set, kit and application for amplifying Salmonella. Background technique [0002] Salmonellosis (Salmonellosis) is a public health disease caused by Salmonella (Salmonella) that is rampant worldwide. Its main symptoms are diarrhea, fever, headache, abdominal cramps and vomiting. Salmonella is the most common foodborne pathogen in my country. Worldwide, almost all ready-to-eat foods and most process standards list Salmonella as a limit indicator. Among the food poisoning caused by bacteria in my country, 70%-80% are caused by Salmonella. It is mainly transmitted by meat and egg products, and can be preserved for 5 months to 2 years in the environment such as feces, soil and water. [0003] The current treatment of Salmonella is mainly through antibiotics. According to literature reports, many Salmonella strains resistant to various antib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/10C12N15/11
CPCC12Q1/689C12Q1/6844C12Q2531/119C12Q2537/1376C12Q2563/107Y02A50/30
Inventor 杜昱光毛瑞梁组源刘雪连刘滢孙明
Owner ZHONGKE XINRAY SUZHOU BIOLOGICAL TECH CO LTD
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