Kit capable of detecting 10 pathogenic vibrios simultaneously and detection method

A technology of pathogenic Vibrio and kits, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc., can solve the problem of small number of pathogenic Vibrio, low efficiency, long cycle, etc. problem, to achieve the effect of shortening the detection cycle and improving detection efficiency

Active Publication Date: 2016-03-16
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, provide a kind of test kit that can detect 10 kinds of pathogenic Vibrio simultaneously and utilize fluorescent probe melting curve method to detect 10 kinds of pathogenic Vibrio simultaneously using the kit of the present invention Bacteria method, aiming to solve the technical problems of small number, low efficiency and long period of detection of pathogenic Vibrio while existing methods for detecting pathogenic Vibrio

Method used

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  • Kit capable of detecting 10 pathogenic vibrios simultaneously and detection method
  • Kit capable of detecting 10 pathogenic vibrios simultaneously and detection method
  • Kit capable of detecting 10 pathogenic vibrios simultaneously and detection method

Examples

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Embodiment 1

[0082] A kit for the simultaneous detection of 10 pathogenic Vibrio species based on the fluorescent probe melting curve method. The kit includes:

[0083] 1. Universal primers, such as QIDNO: 1 and SEQIDNO: 2 in Table 1;

[0084] 2. Fluorescent probes, such as fluorescent probe FAM, fluorescent probe ROX and fluorescent probe Cy5 in Table 1;

[0085] 3. According to the hybrid connection probe of 10 kinds of pathogenic vibrio specific gene design, wherein, 10 kinds of pathogenic vibrio, each vibrio specific gene and each vibrio hybridization connection probe sequence are as in table 2 respectively shown.

Embodiment 2

[0087] A kit for the simultaneous detection of 10 pathogenic Vibrio species based on the fluorescent probe melting curve method. The kit includes:

[0088] 1. Universal primers, such as QIDNO: 1 and SEQIDNO: 2 in Table 1;

[0089] 2. Fluorescent probes, such as fluorescent probe FAM, fluorescent probe ROX and fluorescent probe Cy5 in Table 1;

[0090] 3. According to the hybrid connection probe of 10 kinds of pathogenic vibrio specific gene design, wherein, 10 kinds of pathogenic vibrio, each vibrio specific gene and each vibrio hybridization connection probe sequence are as in table 2 respectively shown.

[0091] 4. The reagent system for the hybridization ligation reaction, as shown in Table 3 above;

[0092] 5. Fluorescent probe melting curve detection reagent system, as shown in Table 4 above.

Embodiment 3

[0094] A kit for the simultaneous detection of 10 pathogenic Vibrio species based on the fluorescent probe melting curve method. The kit includes:

[0095] 1. Universal primers, such as QIDNO: 1 and SEQIDNO: 2 in Table 1;

[0096] 2. Fluorescent probes, such as fluorescent probe FAM, fluorescent probe ROX and fluorescent probe Cy5 in Table 1;

[0097] 3. according to the hybrid connection probe of 10 kinds of pathogenic vibrio specific gene designs, wherein, 10 kinds of pathogenic vibrio, each vibrio specific gene and each vibrio hybrid connection probe sequence are as in table 5 respectively shown.

[0098] 4. The reagent system for the hybridization ligation reaction, as shown in Table 3 above;

[0099] 5. Fluorescent probe melting curve detection reagent system, as shown in Table 4 above.

[0100] Table 510 kinds of pathogenic vibrio hybridization junction probe sequences

[0101]

[0102]

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Abstract

The invention provides a kit capable of detecting 10 pathogenic vibrios simultaneously based on a fluorescence probe melting curve method and a method capable of detecting the 10 pathogenic vibrios simultaneously. The kit comprises a universal primer, a fluorescence probe and hybridization connecting probes designed according to specific genes of the 10 pathogenic vibrios. The method capable of detecting the 10 pathogenic vibrios simultaneously includes the steps that the DNA of dubious bacteria extracted by a sample to be detected is used as a template, and hybridization connection reacting, fluorescence PCR amplification treatment and melting curve analysis treatment are carried out in sequence through the kit by means of the fluorescence probe melting curve method. According to the kit and the detection method, based on the multiple-connecting probe amplification technology, the 10 pathogenic vibrios can be detected simultaneously through the real-time fluorescence PCR melting curve method, hence, the detection period of the pathogenic vibrios is effectively shortened, and the detection efficiency of the pathogenic vibrios is improved.

Description

technical field [0001] The invention belongs to the technical field of sanitation and prevention virus detection, and specifically relates to a kit for simultaneously detecting 10 kinds of pathogenic Vibrio based on a fluorescent probe melting curve method and simultaneously detecting 10 kinds of pathogenic Vibrio based on a fluorescent probe melting curve method. Vibrio method. Background technique [0002] Vibrioceae (Vibrionaceae) is a group of oxidase-positive, short, straight or slightly curved Gram-negative bacilli with terminal flagella. Divided into five genera of Vibrio, Aeromonas, Pseudomonas, Photobacterium and Photobacterium. The genus Vibrio is widely distributed, mostly in water, and there are about 37 species. Among the many vibrios, the pathogenicity is quite different, and the vibrios that are closely related to human health and can cause diseases mainly include: Vibrio cholerae, Vibrio vulnificus, Vibrio river, Vibrio parahaemolyticus, Vibrio mimicus, Vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 扈庆华姜伊祥石晓路李迎慧江敏邱亚群林一曼陈琼成
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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