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Nanogold for detecting Aphelenchoides besseyi and application thereof

A technology of angiostrongylus and nano-gold, which is applied in the field of nano-gold for detecting angiostrongylus cantonensis, can solve problems such as inability to achieve early detection, achieve good early detection and sensitivity, good detection result accuracy, and detection efficiency high effect

Pending Publication Date: 2019-05-21
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

IgM appears about 1 week after infection of stage III larvae, and IgG appears later, this method cannot achieve early detection

Method used

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  • Nanogold for detecting Aphelenchoides besseyi and application thereof
  • Nanogold for detecting Aphelenchoides besseyi and application thereof
  • Nanogold for detecting Aphelenchoides besseyi and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Preparation of Canton round line antigen.

[0088] Animal model establishment

[0089] Infection of intermediate hosts: Feed stage I larvae isolated from feces to apple snails fasted for 24 hours for continuous infection for 3 days, each time feeding for 11 hours (10-12 hours), and eating after 12 hours after infection. For details, please refer to the attached figure 1 .

[0090] Stage III larvae collection: 12 hours after stage I larvae were infected with apple snails, pepsin digestion was used to separate and collect stage III infection stage larvae, crush and remove the reared apple snails, and use a high-speed tissue mixer to fully crush them and weigh them. Add 10g of net weight snail meat with 250mL of digestive juice (3g of pepsin, 7mL of concentrated hydrochloric acid, and 1L of deionized water), put it in a thermostat at 37°C for 10h (8-10h), filter it with a 200-mesh sieve, and filter the obtained Leave the filtrate at room temperature for 15 minutes to pre...

Embodiment 2

[0113] A method for preparing gold nanorods for detecting Angiostrongylus cantonensis.

[0114] Preparation of nano gold rods (seed crystal method)

[0115] Prepare gold crystal seed solution: mix 140 μL HAuCl 4 (0.1M) and 6.86mL of pure water to prepare HAuCl 4 (2mM) solution; freshly prepare NaBH 4 (0.01M) Store at 4°C for 8 minutes; mix 625μL HAuCl 4 (2mM), 1.88mL CTAB (0.2mM, Hexadecyl trimethyl ammonium Bromide, hexadecyltrimethylammonium bromide) and 1.37mL pure water, mix well and stir for 1 minute, then add freshly prepared 450μL NaBH 4 Obtain the golden crystal seed liquid, shake vigorously for 2 minutes, and the color of the golden crystal seed liquid changes from yellow to light brown. Place in a 27°C incubator and incubate for 2 hours to obtain a gold crystal seed solution.

[0116] Prepare growth medium: mix CTAB (0.2M, 11.875mL), 7.71mL pure water, HAuCl 4 (2M, 5mL) shake well, then add AgNO 3 (10mM, 150μL), AA (100mM, 160μL, ascorbic acid) were oscillated...

Embodiment 3

[0133] The optimal coating concentration of the antigen provided in Example 2 was verified.

[0134] Ultraviolet spectrophotometer was used to detect the shift of the red peak of the longitudinal plasmon resonance band before and after the antigen was labeled with gold nanorods, and the optimal coating concentration of the antigen was determined according to the shift of the red peak.

[0135] Experimental results

[0136] V stage larval crude antigen

[0137] The scanning spectrum of ultraviolet spectrophotometer (UV-Vis) is attached Figure 9 As shown, the changes of the longitudinal plasmon resonance band coating and detection absorption peak (LSPR) are as attached Figure 10 shown.

[0138] attached by Figure 10 It can be seen that according to the shift of the red peak, the optimal dilution concentration of stage V larval antigen is: 30 μg / mL>50 μg / mL>10 μg / mL=20 μg / mL=40 μg / mL>60 μg / mL. Therefore, when the maximum red peak shift of V-stage larvae before and after c...

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Abstract

The invention discloses nanogold for detecting Aphelenchoides besseyi and application thereof, and relates to the technical field of Aphelenchoides besseyi detection. Specifically, the nanogold is prepared by coating a nanogold matrix with Aphelenchoides besseyi antigen, and the nanogold can effectively detect the Aphelenchoides besseyi, has good detection early-stage property and sensitivity, andis high in detection efficiency and good in detection result precision.

Description

technical field [0001] The invention relates to the technical field of detection of Angiostrongylus cantonensis, in particular to a nano-gold for detecting Angiostrongylus cantonensis and its application. Background technique [0002] Angiostrongylus cantonensis (Angiostrongylus cantonensis) is a food-borne zoonotic parasitic disease first discovered by Professor Chen Xintao in the lungs of Rattus norvegicus and black mice in Guangzhou. It is also known as eosinophilic meninges inflammation or meningoencephalitis. In terms of classification, it belongs to the phylum Nematozoa, the class Caudosensilla, the order Protostrata, the family Angiostrongylaceae, and the genus Angiostrongylus. The ultimate host of the worm is rodents, mainly Rattus norvegicus and house rat. [0003] The primary larvae were isolated from the feces of rodents infected with the final host of Angiostrongylus cantonensis, and infect the intermediate host molluscs, mainly apple snails, brown cloud snails...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/68G01N33/532
Inventor 杨毅梅吕跃军巫秀美孔玉芳刘衡
Owner DALI UNIV
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