GB/T 34750-2017 in English

1. Background of Standard Formulation and Analysis of Technological Evolution

Haemophilus parasuis is a pathogen that seriously affects the pig farming industry and can cause a variety of diseases in pigs, such as sepsis, arthritis and meningitis. In order to standardize its detection methods, the Ministry of Agriculture of the People's Republic of China organized the drafting of the GB/T 34750—2017 standard, which combines isolation and culture, PCR amplification and real-time fluorescence quantitative PCR technology to provide a standardized process for laboratory diagnosis.

2. Comparative analysis of standard frameworks

3. Laboratory testing implementation recommendations

Sample collection and processing: According to the standard requirements, the storage conditions (2-8℃) of the samples after collection must be strictly controlled and sent to the laboratory within 24 hours. For tissue samples, it is recommended to use sterile scissors and grinders for processing, and add phosphate buffered saline (PBS) to mix.

Precautions for nucleic acid extraction: The phenol-chloroform-isoamyl alcohol mixture is toxic, and protective equipment must be worn during operation. During DNA extraction, care should be taken to avoid RNA contamination. It is recommended to use commercial kits or standardized processes.

PCR detection optimization: Nested PCR and real-time fluorescence PCR both require strict control of reaction conditions (such as annealing temperature, number of cycles, etc.). It is recommended that laboratories establish a quality control system for negative and positive controls and calibrate instruments and equipment regularly.