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Toxoplasma detection kit

A detection kit and Toxoplasma gondii technology, applied in the biological field, can solve the problems of unsuitable on-site detection and long-term detection, and achieve the effects of easy promotion, high sensitivity and large expression

Active Publication Date: 2016-07-06
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods take a long time and require some equipment or professionally trained technicians, so they are not suitable for large-scale on-site detection.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment one kit preparation

[0027] The kit of the present invention comprises: test strips, standard products, reference products, and sample diluent; wherein the standard product is Toxoplasma gondii positive serum, the reference product is Toxoplasma gondii negative serum, and the sample diluent is 10mM, pH7.5 Phosphate buffered saline (PB), consisting of NaH 2 PO 4 ·H 2 O1.38g, Na 2 HPO 4 12H 2 O3.58g, add double distilled water to 100ml, adjust the pH to 7.5.

[0028] The kit of the present invention is prepared through the following steps:

[0029] (1) Preparation of anti-SAG3 monoclonal antibody: 8-10 weeks old BALB / c mice were taken, and each mouse was intraperitoneally injected with 1 mL of paraffin oil; 7 days later, each mouse was intraperitoneally injected with 1 x 10 6 A monoclonal hybridoma cell line 4D5 (preserved in the China Center for Type Culture Collection, preservation number CCTCCNO: C201624, preservation date: 2016.2.1), ascites was co...

Embodiment 2

[0037] Purification of Example Secondary Anti-Toxoplasma gondii SAG3 Monoclonal Antibody

[0038] 1. Materials

[0039] Monoclonal antibody ascites was preserved by our laboratory; reagents such as ammonium sulfate and n-octanoic acid were of domestic analytical grade; the freeze dryer and microplate reader were purchased from Thermo Company; the centrifuge was purchased from HITACHI.

[0040] 2. Monoclonal Antibody Purification

[0041] Monoclonal antibody was purified by octanoic acid-ammonium sulfate precipitation method. First, take out ascites from the refrigerator, put it on ice to melt, then centrifuge at 12000rpm for 30min, take the supernatant, and discard the precipitated impurities; mix the ascites and acetate buffer according to 1 : 2, mixed at room temperature according to 33μl / ml ascites, added n-octanoic acid drop by drop, stirred at room temperature for 30min, and overnight at 4°C to make it fully precipitated; under the condition of 4°C, centrifuged at 12000r...

Embodiment 3

[0046] Example 3 Performance Evaluation of Colloidal Gold Immunochromatography Rapid Detection Test Strip

[0047] 1. Materials

[0048] Toxoplasma gondii positive serum, Cryptosporidium pig positive serum, Streptococcus suis positive serum, Salmonella pig positive serum, Cysticercus pig positive serum, Coccidia pig positive serum, Eperythrozoon pig positive serum, Trichinella pig positive serum Toxoplasma gondii Natural whole protein, Toxoplasma gondii negative serum is preserved by our laboratory.

[0049] 2. Specificity test

[0050] The colloidal gold immunochromatographic rapid detection test strip detection method established in this study was used to detect porcine Toxoplasma gondii positive serum, Porcine Cryptosporidium positive serum, Streptococcus suis positive serum, Porcine Salmonella positive serum, Cysticercus porcine positive serum, porcine Coccidia positive serum, porcine Eperythrozoon positive serum, porcine Trichinella positive serum, in order to determine...

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Abstract

The invention provides a monoclonal hybridoma cell strain 4D5.The collection number is CCTCC NO: C201624, the classification name is hybridoma cell strain No.4D5, and the collection date is February 1st, 2016.The invention further provides a toxoplasma detection kit.The toxoplasma detection kit is formed by a test strip, a standard sample, a reference sample and a sample diluent.An anti-toxoplasma SAG3 monoclonal monomer embedded in the test strip is secreted by the monoclonal hybridoma cell strain 4D5 with the collection number CCTCC NO: C201624.According to the toxoplasma detection kit, the immunochromatographic strip technology is adopted, and the technology is short in detection time, low in cost, easy to operate and suitable for field detection and epidemiological surveying; toxoplasma SAG3 serves as a diagnostic antigen; compared with SAG1 and SAG2, the expression quantity of the SAG3 is large, the toxoplasma SAG3 can be expressed in all infection stages of toxoplasma and more suitable for serving as the diagnostic antigen; the clinical application result shows that the toxoplasma detection kit is high in sensibility and specificity, good in stability and easy to popularize to the primary level.

Description

technical field [0001] The invention belongs to biotechnology, and relates to a detection kit based on colloidal gold immunochromatography rapid detection technology of toxoplasma surface protein 3 (SAG3), which can rapidly detect toxoplasma infection in humans, pigs, cats, dogs and the like. Background technique [0002] Toxoplasmosis is a worldwide zoonotic parasitic disease caused by the opportunistic intracellular parasite Toxoplasma gondii. This parasite can invade all warm-blooded animals, including humans. The body with normal immunity will not show obvious symptoms, but it will cause hydrocephalus, retinitis, choroiditis, encephalitis in people with low immunity, and even death in people with impaired immune system. Toxoplasmosis also has a major impact on the livestock industry, causing abortions, stillbirths and deaths of newborns, especially in sheep and goats. Therefore, the status of toxoplasmosis in public health and safe production of livestock and poultry i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/54306G01N33/558
Inventor 杜爱芳罗嘉庆卓洵辉孙洪超杨怡陈学秋
Owner ZHEJIANG UNIV
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