Method for early detection of grouper intestinal tract microsporidia and application thereof
A technology for early detection of microsporidia, applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, can solve the problem of losing the best time for treatment, delaying treatment, and loss of grouper aquaculture and other issues, to achieve the effects of programming and standardization, detection specificity and high sensitivity
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[0024] The early detection method of embodiment 1 grouper intestinal microsporidia
[0025] (1) Select about 1 cm of grouper foregut, scrape the intestinal contents, store in 90% absolute alcohol or directly use for the extraction of genomic DNA;
[0026] (2) Use conventional phenol chloroform or commercial fecal sample genome extraction kit to extract genomic DNA from intestinal contents;
[0027] (3) Using the extracted genomic DNA as a template, design and synthesize specific primers for PCR amplification;
[0028] EntF:5'-GACGTGACTAAAAGAGCGT-3';
[0029] EntR: 5'-AAGTTTCAGTCTTGCGGCT-3'.
[0030] PCR reaction system (25ul):
[0031]
[0032] PCR reaction program:
[0033] 95℃*4min,
[0034]
[0035] 72℃*10min
[0036] After the reaction, turn on the PCR instrument, take out the amplified sample, and store it at 4°C.
[0037] (4) Then take an appropriate amount of the PCR amplification product of step 3) and separate it by agarose electrophoresis, observe it und...
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