Detection gene chip and detection method of main pathogenic bacteria of dairy cow mastitis

Abstract

The invention provides a detection gene chip and a detection method for the main pathogenic bacteria of cow mastitis. The gene chip includes a solid phase carrier and a specific detection probe fixed on the solid phase carrier, and the specific detection probe includes 6 kinds of universal primer sequences designed with the Escherichia coli 16S rDNA conserved region as the base sequence. A specific oligonucleotide hybridization probe designed in the variable region of the main pathogenic bacteria of dairy cow mastitis; the six main pathogenic bacteria of dairy cow mastitis are Streptococcus agalactiae, Streptococcus dysgalactiae, Escherichia coli, pneumoniae Klebsiella, Proteus mirabilis, and Staphylococcus aureus. The invention precisely limits the annealing temperature of the PCR reaction, optimizes the hybridization conditions of the PCR-amplified nucleic acid fragment and the gene chip, obtains a good hybridization effect, and finally obtains accurate detection results. The invention has good specificity, high accuracy rate, reliable stability, low false positive rate of detection results, simple operation steps and easy popularization and use.

Description

technical field [0001] The invention belongs to the technical field of detection and relates to a rapid detection technology, in particular to a detection gene chip and a detection method for the main pathogenic bacteria of cow mastitis. Background technique [0002] Cow mastitis not only seriously affects the quality of milk, causes the productivity of dairy cows to drop seriously and even be eliminated, but also endangers human health. The epidemic of dairy cow mastitis has become a recognized public health, social and economic problem. There are many types of pathogenic bacteria that cause mastitis in dairy cows. At present, there are more than 220 known pathogenic bacteria, of which more than 20 are common, and 90% of the pathogenic bacteria of cow mastitis are mainly Escherichia coli, Staphylococcus aureus Coccus, Staphylococcus epidermidis, Klebsiella pneumoniae, Proteus mirabilis, Streptococcus dysgalactiae, Streptococcus agalactiae, Streptococcus uberis, Salmonella, ...

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Problems solved by technology

[0003] Chinese patent application 200710166530.3 discusses the detection gene chip of pathogenic bacteria in milk products, fluorescently labeled on the probe, the probe is obtained by screening between 16S-23S rDNA, but the interval is only more than 200 bases, It is very difficult to screen out 10 probes, and cannot be applied to the accurate detection of the main pathogenic bacteria of cow mastitis

However, no relevant technical reports and solutions have been seen so far

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