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Method for indirect immunofluorescence assay (IFA) detection of eperythrozoonosis by use of monoclonal antibody

A technology for immunofluorescence detection and eperythrozoonosis, which is used in fluorescence/phosphorescence, measurement devices, disease diagnosis, etc., to achieve the effects of strong sensitivity, high specificity, and high accuracy and reliability

Inactive Publication Date: 2014-06-18
QINGDAO ZHONGREN PHARMA
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  • Application Information

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Problems solved by technology

Molecular biological detection methods developed in recent years, such as PCR (polymerase chain reaction), nucleic acid hybridization, etc., have begun to be applied to the detection and diagnosis of this disease, but specific and sensitive detection methods need to be improved urgently, and the probability of false negative and false positive Very big

Method used

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Embodiment

[0022] The method for detecting Eperythrozoonosis by monoclonal antibody indirect immunofluorescence in this embodiment comprises the following steps:

[0023] (1) The establishment of the standard positive control and negative control, the pig blood with a natural infection rate of more than 90% and positive by Wright's, Giemsa and acridine orange staining was used as the standard positive antigen control; The pig blood in the area that was negative for compression, Wright's, Giemsa, and acridine orange staining and PCR was used as a negative control;

[0024] (2) For antigen coating, take standard positive and negative blood smears, dry them, and set a blank control;

[0025] (3) Dilute the fluorescent secondary antibody, dilute the FITC-labeled goat anti-mouse IgG antibody with 0.01% Evans blue and 0.01 mol / L PBS respectively;

[0026] (4) For IFA determination, fix the coated two sets of standard positive and negative control slides with a mixture of equal proportions of ...

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Abstract

The invention discloses a method for indirect immunofluorescence assay (IFA) detection of eperythrozoonosis by use of a monoclonal antibody. The method includes establishment of a standard positive control and a standard negative control, antigen coating, fluorescence second antibody dilution, IFA determination and other steps. An antibody used in the method is a monoclonal antibody against porcine eperythrozoon, and compared with general antibodies used in IHA (indirect hemagglutination assay), ELISA (enzyme-linked immuno sorbent assay) and other detection methods, the antibody used in the method has the advantages of high specificity, high sensitivity, high accuracy and reliability, can fast, simply, reliably and accurately detect eperythrozoon in a sample, and can also be used for epidemiological investigation and curative effect detection of the eperythrozoonosis.

Description

[0001] technical field [0002] The invention belongs to the technical field of bioengineering detection, and in particular relates to a method for detecting eperythrozoonosis by monoclonal antibody indirect immunofluorescence. [0003] Background technique [0004] Eperythrozoonsis (Eperythrozoonsis) is a zoonotic disease with anemia, jaundice and fever as the main clinical symptoms caused by Eperythrozoon (EH) parasitizing on the surface of human and animal red blood cells, plasma and bone marrow. . Belongs to Rickettsiales (Rickettsiales), Anaplasmata-ceae (Anaplasmata-ceae), Eperythrozoon (Eperythrozoon). Most of them are single or clustered on the surface of red blood cells, in the form of chains or scales, and a small amount of them are free. Porcine Eperythrozoon is generally ring-shaped, with a diameter of 0.8-2.5um, and there are also spherical and rod-shaped. It is sensitive to drying and chemical drugs. 0.5% carbolic acid can kill it after 3 hours at 37 ° C. G...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/64
CPCG01N33/577G01N33/54373G01N2800/22
Inventor 张述智夏伟朱绍辉张浩王晓丽徐权汗李之详许团辉
Owner QINGDAO ZHONGREN PHARMA
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