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Salmonella and shigella joint detection kit and detection method thereof

A technology for Salmonella and Shigella, applied in biochemical equipment and methods, microbe measurement/inspection, and resistance to vector-borne diseases, etc., can solve the problems of long time-consuming, low missed detection rate, and high missed detection rate, and achieve The effect of high yield, high accuracy and high verification rate

Active Publication Date: 2012-09-19
GUANGZHOU HUAFENG BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] One of the technical problems to be solved by the present invention is to overcome the shortcomings of long time consumption and high missed detection rate in the prior art to provide a Salmonella and Shigella combined detection kit with more comprehensive detection effect, high specificity and low missed detection rate

Method used

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  • Salmonella and shigella joint detection kit and detection method thereof
  • Salmonella and shigella joint detection kit and detection method thereof
  • Salmonella and shigella joint detection kit and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0065] Kit preparation

[0066] (1) Synthesize oligodeoxynucleic acid primers by DNA synthesizer according to the following sequence:

[0067] Salmonella

[0068] AgfA-9-F3: AGTTTCTGGCAGTGCTGTG (SEQ ID NO 1)

[0069] AgfA-9-B3: AGTACTGTTATCCGCACCCT (SEQ ID NO 2)

[0070] AgfA-9-FIP: ATCCGGGCCGGAACTATTGCTTTTCTGGCGTCGTTCCACAAT (SEQ ID NO 3)

[0071] AgfA-9-BIP: CGCTTGCTCTGCAAAGCGATGTTTTACCATAACCGCTCTGGGT (SEQ ID NO 4)

[0072] Shigella

[0073] ipaH-69-F3: ACATGAAGAGCAYGCCAACA Y represents C or T (SEQ ID NO 17)

[0074] ipaH-69-B3: TCCTCCAGCTCTCAGTGG (SEQ ID NO 18)

[0075] ipaH-69-FIP: CGGAATCCGGAGGTATTGCGTGTTTTCCTTTTCCGCGTTCCTTGA (SEQ ID NO 19)

[0076] ipaH-69-BIP: GGTCGCTGCATGGCTGGAAATTTTGCAGCAACAGCGAAAGACT (SEQ ID NO 20)

[0077] (2) Purchase DNA polymerase: Bst DNA polymerase is placed in the container;

[0078] (3) Prepare the buffer: the buffer is 0.18mol / L Tris-HCl, 0.10mol / L KCl, 0.07mol / L (NH 4 ) 2 SO 4 , 15mmol / L MgSO 4 , prepared with 1 volume % TritonX...

Embodiment 2

[0093] Kit preparation

[0094](1) Synthesize oligodeoxynucleic acid primers by DNA synthesizer according to the following sequence:

[0095] Salmonella

[0096] AgfA-56-F3: CCCGGATTCCACGTTGAG (SEQ ID NO 5)

[0097] AgfA-56-B3: TCGGAGTTTTTAGCGTTCCA (SEQ ID NO 6)

[0098] AgfA-56-FIP: CCGCTCTGGGTAATGGTCGTTTTTTTCTAACGCTGCGCTTGCTC (SEQ ID NO 7)

[0099] AgfA-56-BIP: ATGTAGGCCAGGGTGCGGATATTTTGGTCGATGGTGGCATTG (SEQ ID NO 8)

[0100] Shigella

[0101] ipaH-69-F3: ACATGAAGAGCAYGCCAACA Y represents C or T (SEQ ID NO 17)

[0102] ipaH-69-B3: TCCTCCAGCTCTCAGTGG (SEQ ID NO 18)

[0103] ipaH-69-FIP: CGGAATCCGGAGGTATTGCGTGTTTTCCTTTTCCGCGTTCCTTGA (SEQ ID NO 19)

[0104] ipaH-69-BIP: GGTCGCTGCATGGCTGGAAATTTTGCAGCAACAGCGAAAGACT (SEQ ID NO 20)

[0105] (2) Purchase DNA polymerase: Bst DNA polymerase is placed in the container;

[0106] (3) Prepare the buffer: the buffer is 0.25mol / L Tris-HCl, 0.15mol / L KCl, 0.15mol / L (NH 4 ) 2 SO 4 , 30mmol / L MgSO 4 , prepared with 2 volume % Trit...

Embodiment 3

[0116] Kit preparation

[0117] (1) Synthesize oligodeoxynucleic acid primers by DNA synthesizer according to the following sequence:

[0118] Salmonella

[0119] AgfA-19-F3: AGTTTCTGGCAGTGCTGTG (SEQ ID NO 9)

[0120] AgfA-19-B3: AGTACTGTTATCCGCACCCT (SEQ ID NO 10)

[0121] AgfA-19-FIP: ATCCGGGCCGGAACTATTGCCTGGCGTCGTTCCACAAT (SEQ ID NO 11)

[0122] AgfA-19-BIP: CGCTTGCTCTGCAAAGCGATG ACCATAACCGCTCTGGGT (SEQ ID NO 12)

[0123] Shigella

[0124] ipaH-69-F3: ACATGAAGAGCAYGCCAACA Y represents C or T (SEQ ID NO 17)

[0125] ipaH-69-B3: TCCTCCAGCTCTCAGTGG (SEQ ID NO 18)

[0126] ipaH-69-FIP: CGGAATCCGGAGGTATTGCGTGTTTTCCTTTTCCGCGTTCCTTGA (SEQ ID NO 19)

[0127] ipaH-69-BIP: GGTCGCTGCATGGCTGGAAATTTTGCAGCAACAGCGAAAGACT (SEQ ID NO 20)

[0128] (2) Purchase DNA polymerase: Bst DNA polymerase is placed in the container;

[0129] (3) Prepare the buffer: the buffer is 0.2mol / L Tris-HCl, 0.1mol / L KCl, 0.1mol / L (NH 4 ) 2 SO 4 , 20mmol / L MgSO 4 , prepared with 1 volume % TritonX-10...

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Abstract

The invention relates to a salmonella and shigella joint detection kit and a detection method thereof. The kit of the invention comprises Bst DNA polymerase, buffer solution, dNTPs, betaine, magnesium sulfate, development solution, stabilizing solution and positive control; and the kit also comprises two pairs of primers, namely inner primers FIP / BIP and outer primers F3 / B3 which take salmonella AgfA genes and shigella ipaH genes as target genes and are designed based on loop-mediated thermostatic amplification technology. The salmonella and shigella detection kit has the advantages of more comprehensive detection effect, high specificity and low omission factor, and is suitable for quickly detecting salmonella and shigella in physical examination of food employees.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic reagents, in particular to a combined detection kit for Salmonella and Shigella and a detection method thereof. Background technique [0002] Salmonella, Enterobacteriaceae, Gram-negative enterobacteriaceae. Salmonellosis is one of the zoonotic diseases of great significance in public health and epidemiology, and is one of the main intestinal pathogenic bacteria, which can easily cause food poisoning. Humans and animals can be asymptomatic carriers after infection, and can also manifest as symptoms. A clinically lethal disease that may increase morbidity or mortality, or reduce the reproductive productivity of an animal. In my country, food poisoning caused by Salmonella occupies the first place in bacterial food poisoning. Salmonella usually enters the human body through ingestion of food contaminated with animal feces. The food that is generally susceptible to contamination is mai...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/10
CPCY02A50/30
Inventor 章琪陆晔田桢干方筠张晓航王健张琳王传现曹以诚陈洵杜正平谭慧媚
Owner GUANGZHOU HUAFENG BIOTECH
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