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Detection kit, detection method and application for shigella

A detection kit, Shigella technology, applied in the biological field, can solve the problems of expensive instruments, complex detection procedures, and complicated primer design, and achieve the effects of short reaction time and easy operation.

Active Publication Date: 2020-04-14
INSPECTION & QUARANTINE TESTING CENT OF HEBEI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the existing problems such as complex detection procedure of Shigella, tedious design of primers, expensive instruments, etc., the present invention provides a detection kit for Shigella

Method used

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  • Detection kit, detection method and application for shigella
  • Detection kit, detection method and application for shigella
  • Detection kit, detection method and application for shigella

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Experimental program
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Embodiment 1

[0045] Embodiment 1 is used for the detection method of Shigella-establishment of real-time fluorescence RPA method

[0046] 1. Design and preparation of primers and probe sequences

[0047] In this study, the Shigella invasion plasmid antigen H gene (ipaH) was used as the target gene, specific primers and exo probes were designed according to its conserved sequence, and a real-time RPA for the rapid detection of Shigella in food was established. method.

[0048] The inventors of the present invention analyzed the Shigella ipaH gene sequence in GeneBank (accession number: AE005674), selected the conserved region, and designed real-time RPA primers and exo probes. The target fragment size was 247bp.

[0049] The present invention designed and synthesized a pair of primers and a probe, as shown in Table 1.

[0050] Table 1 Shigella real-time RPA primers and probes designed by the present invention

[0051]

[0052] 2. Extraction of Shigella strains and bacterial genomic DN...

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Abstract

The invention relates to the field of biotechnology, and specifically discloses a detection kit, a detection method and an application for Shigella. The detection kit is used for real-time fluorescence RPA (recombinase polymerase amplification) detection of Shigella; and the detection method uses a Shigella ipaH gene as a target gene to establish a real-time fluorescence RPA method for effectively detecting Shigella. The method is strong in specificity and high in sensitivity, has a detection limit of 3.5*10<-3>ng / muL, only needs 20min to complete the detection at a constant temperature of 39 DEG C, and has a detection time of only 7-12min, so that the Shigella in a food sample can be effectively detected, the method can be applied to field detection, and the novel method is provided for detecting the Shigella in food and food-borne public health events.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit, detection method and application for Shigella. Background technique [0002] Food-borne diseases are currently an important public health issue of concern to the whole world. No matter in developed or developing countries, tens of millions of people are infected with food-borne pathogens every year, and severe cases lead to death. As one of the main foodborne pathogens, Shigella ranks first among the pathogens causing infectious diarrhea in my country. The genus Shigella consists of four Gram-negative, non-motile, non-spore-forming rod-shaped bacteria, Shigella baumannii, Shigella dysenteriae, Shigella flexneri, and Sonnei Shigella spp. The more virulent species of Shigella can cause bacillary dysentery in humans, which can present with mild dysentery, fever, abdominal cramps, and severe fluid loss. [0003] The current molecular biology detection methods can effe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6844C12Q1/689C12Q1/10C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2521/507C12Q2561/113C12Q2563/107
Inventor 王建昌刘立兵孙晓霞娄巧哲南汇珠
Owner INSPECTION & QUARANTINE TESTING CENT OF HEBEI ENTRY EXIT INSPECTION & QUARANTINE BUREAU