Primer and probe for detecting vibrio vulnificus and detection method using the same

A technology of Vibrio vulnificus and probe, applied in the field of identification, detection, or quantification of Vibrio vulnificus, can solve the problems of sufficient specificity, no hemolysin gene comparison, and specificity has been fully proved.

Inactive Publication Date: 2005-10-05
NICHIREI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, based on the primers designed, no comparison was made to these hemolysin genes
Therefore, it cannot be said that specificity has been sufficiently demonstrated
[0007] As mentioned above, it cannot be said that the specificity is sufficient with regard to the existing methods for detecting Vibrio vulnificus using genes

Method used

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  • Primer and probe for detecting vibrio vulnificus and detection method using the same
  • Primer and probe for detecting vibrio vulnificus and detection method using the same
  • Primer and probe for detecting vibrio vulnificus and detection method using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] An example of gene amplification using primers designed and obtained using a region specific to Vibrio vulnificus according to the present invention is shown.

[0032] target gene

Primer

sequence

length

site a)

direction

gyrB

VF1

VR1

5′-gatgcaccgcttgctatcatc-3′

5′-ttgtctgccatgaaggattcc-3′

21

21

121-141

740-720

justice

antonym

wxya

VrF2

VrR2

5′-gaactgatgctcgatgtgttt-3′

5′-ttgatgttgytyactgaaagc-3′

21

21

442-462

770-750

justice

antonym

recA

VVrecF2

VVrecR2

5′-cctgtgtatgcgaagaarctt-3′

5′-tcaaccgcmcctgagcgagca-3′

21

21

133-153

248-228

justice

antonym

[0033] a): Indicates the position from the 5' end of the nucleotide sequence represented by SEQ ID NOS: 1-3.

[0034] In addition, the primers described in clai...

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PUM

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Abstract

To produce a high-performance specific gene amplification primer for detecting, quantifying, or identifying Vibrio vulnificus, having low risk of misidentification and practically sufficient amplification efficiency and amplification specificity. We have determined partial nucleotide sequences of gyrB, rpoD, and recA genes of Vibrio vulnificus and the closely related species, revealed their phylogenetic relationship, and then identified nucleotides characteristic to Vibrio vulnificus. Thus, we have made it possible to design a probe having high specificity and a gene amplification primer having high specificity and excellent amplification efficiency, both of which contain the characteristic nucleotides.

Description

technical field [0001] The invention relates to a method for detecting, identifying, or quantifying Vibrio vulnificus in food detection, epidemiological environment detection, and clinical inspection. Background technique [0002] Vibrio vulnificus is a halophilic Gram-negative bacterium found in seawater and brackish water areas. Although rarely infecting healthy humans, Vibrio vulnificus infects people with serious primary diseases such as liver disease. cirrhosis of patients and can cause severe infectious disease. As an infectious bacterium associated with fatal cases, it has attracted attention in public health. The first case reports of infectious diseases produced by this bacterium include by Roldan in 1970 ( New Engl.J.Med., 282,1306,1970) isolated bacteria from lower gangrene. This case was initially considered to be a case of extraintestinal infection of Vibrio rahaemolyticus. Since then, it has been revealed that this bacterium degrades lactose more than large int...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N9/90C12N15/31C12Q1/68
CPCC12Q1/689C12N9/90C12Y599/01003Y02A50/30
Inventor 小泉雄史西山叶子山本敏福山正文吉畑胜则大仲贤二
Owner NICHIREI CORP
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