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Primer set, probe, kit and detection method for detecting schistosoma japonicum intermediate host oncomelania

An intermediate host and detection method technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of long detection time, cumbersome operation, false positive results, etc., and achieve low cost, The effect of avoiding missed detection

Active Publication Date: 2020-11-20
JIANGSU INST OF PARASITIC DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Based on this, it is necessary to address the technical problems of long detection time, cumbersome operation and false positive results of the traditional laboratory nucleic acid detection method for schistosomiasis. The present invention proposes a primer set, probe and Needles, kits and detection methods

Method used

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  • Primer set, probe, kit and detection method for detecting schistosoma japonicum intermediate host oncomelania
  • Primer set, probe, kit and detection method for detecting schistosoma japonicum intermediate host oncomelania
  • Primer set, probe, kit and detection method for detecting schistosoma japonicum intermediate host oncomelania

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The kit composition of Example 1 is shown in Table 1.

[0068] The kit composition of table 1 embodiment 1

[0069]

[0070] Among them, lateral flow chromatography test paper, magnesium acetate solution, PBS buffer solution, and reaction unit tubes can also be provided separately outside the kit.

[0071] Prepare two reaction unit tubes, add a part of the reaction system to each reaction unit tube and mix well (do not use a vortex shaker to shake vigorously to mix well), and after a short centrifugation, collect the liquid of the reaction system to the bottom of the reaction unit tube.

[0072] Add 2.5 μL of magnesium acetate solution to each reaction unit tube, add 1 μL of sterile water and 1 μL of 10 4 Copies / μL of Schistosoma japonicum recombinant plasmid SjG28.

[0073] Use the B6100 constant temperature oscillating mixer equipped with Qitian Gene to oscillate and mix (incubating for 4 minutes), and place the reaction unit tube in a 39°C water bath for isother...

Embodiment 2

[0078] Example 2 The recombinant plasmid SjG28 of Schistosoma japonicum was used as a template to evaluate the sensitivity of the kit and detection method for detecting Schistosoma japonicum intermediate host snails of the present invention.

[0079] The composition of the kit of Example 2 is shown in Table 2.

[0080] The kit composition of table 2 embodiment 2

[0081]

[0082]

[0083] Among them, lateral flow chromatography test paper, magnesium acetate solution, PBS buffer solution, and reaction unit tubes can also be provided separately outside the kit.

[0084] Prepare four reaction unit tubes, add a portion of the reaction system to each reaction unit tube and mix well (do not use a vortex shaker to mix vigorously). After a short centrifugation, collect the liquid of the reaction system to the bottom of the reaction unit tube.

[0085] Add 2.5 μL of magnesium acetate solution to each reaction unit tube, add 1 μL of sterilized water, 1 μL of 10copies / μL of Schist...

Embodiment 3

[0093] Example 3 The DNA extract of Clonorchis sinensis, the DNA extract of Schistosoma mansoni adults, the DNA extract of Ancylostoma duodenale, the DNA extract of Schistosoma haematobium, and the DNA extract of Babesia gibsoni , Paragonimus westermani adult DNA extract as a template to evaluate the specificity of the kit and detection method for detecting Schistosoma japonicum intermediate host snails of the present invention.

[0094] The kit composition of Example 3 is shown in Table 3.

[0095] The kit composition of table 3 embodiment 3

[0096]

[0097]

[0098] Among them, lateral flow chromatography test paper, magnesium acetate solution, PBS buffer solution, and reaction unit tubes can also be provided separately outside the kit.

[0099] Prepare six reaction unit tubes, add a portion of the reaction system to each reaction unit tube and mix well (do not use a vortex shaker to shake vigorously to mix well), after a short centrifugation, collect the liquid of t...

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Abstract

The invention provides a primer set, a probe, a kit and a detection method for detecting schistosoma japonicum intermediate host oncomelania, wherein the primer set comprises an upstream primer and adownstream primer; the upstream primer has a sequence shown as SEQ ID No. 1; and the downstream primer has a sequence shown in SEQ ID No. 2. The primer set, the probe, the kit and the detection methodhave characteristics of rapidness, simplicity, convenience, visibility, no pollution and low cost, can be used for on-site detection and instant detection, and can avoid occurrence of missed detection.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection, in particular to a primer set, probe, kit and detection method for detecting Schistosoma japonicum intermediate host snails. Background technique [0002] Schistosomiasis is a zoonotic parasitic disease that seriously endangers human health and affects economic development. [0003] There are many kinds of final hosts of Schistosoma japonicum. It is reported that more than 40 kinds of mammals of 7 orders, 28 genera and 28 genera are naturally infected with Schistosoma japonicum, including cattle, pigs, goats, sheep, dogs, cats, hares, voles, and horses [6]. Oncomelania is the only intermediate host of Schistosoma japonicum, and plays a very important role in the transmission of schistosomiasis, and its infection status can reflect the epidemic situation of schistosomiasis endemic areas. The life history of schistosomiasis is complex, and the factors affecting the prevalence a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6804C12N15/11
CPCC12Q1/6888C12Q1/6804C12Q2521/507C12Q2522/101C12Q2563/131C12Q2565/625
Inventor 杨坤赵松张键锋叶钰滢
Owner JIANGSU INST OF PARASITIC DISEASES
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