GB/T 34740-2017 in English

Analysis of Standard Formulation Background and Technology Evolution

GB/T 34740—2017 standard was formulated based on the relevant guidelines of the World Organization for Animal Health (OIE) and in combination with China's actual situation. This standard mainly stipulates the specific operation process and judgment criteria of the direct immunofluorescence diagnostic method for animal rabies.

Technical Features

• Rapid Detection: Through fluorescence microscopy observation, the presence of viral antigens can be determined in a short time.
• High Sensitivity: Direct immunofluorescence (DFA) uses FITC-labeled antibodies to significantly improve the sensitivity and specificity of detection.

Comparative analysis of standard frameworks

Experimental operation process and result judgment

Sample collection and preparation

• Small animals: Open the cranial cavity and collect all brain tissues.
• Medium and large animals: Use the foramen magnum sampling method to collect samples from key parts such as the medulla oblongata.

Fluorescent antibody staining steps

1. Touch slide preparation: Take a rice-sized tissue imprint in a biosafety cabinet and dry it at room temperature for 10 minutes.
2. Fixation: Soak in 80% acetone solution for 30 minutes and dry it naturally for 10 minutes.
3. Staining and sealing: Add Evans blue to the fluorescent antibody working solution, incubate, wash and seal the slide.

Result judgment

Positive sample characteristics: bright apple green fluorescent particles.

Implementation recommendations and precautions

Laboratory operation recommendations

• Ensure that reagents and equipment meet standard requirements, especially the sensitivity of the fluorescence microscope.
• Strictly control the time interval from sample collection to testing to avoid corruption affecting the results.

Personnel protection measures

• Operators must complete rabies pre-exposure immunization and regularly test antibody levels.
• Wear double gloves and goggles during the experiment to avoid direct contact with samples.