Nested PCR detection method or/and identification method of brucella
A Brucella, to-be-detected technology, applied in the field of molecular biology, can solve the problems of complex operation process, high false positive rate and false negative rate, difficult detection, etc., and achieve high sensitivity, good specificity, and simple test operation. Effect
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[0048] In the embodiment of the present invention, the nested PCR simultaneously detects and identifies Brucella species and biotypes. First, according to the transposase orfA and transposase orfB gene sequences registered in GenBank, the Primer 5.0 software is used to design the nested PCR first and second respectively. Secondary amplification primers.
[0049] Primers for the first PCR amplification:
[0050] Forward primer: F1: 5'-AACGTAACCAGATCATAGCGCATG-3', as shown in SEQ ID NO: 1;
[0051] Reverse primer: R1: 5'-ACAGATGAGCAATGGAACCGGAT-3', as shown in SEQ ID NO: 2;
[0052] Primers for the second PCR amplification:
[0053] Forward primer: F2: 5'-GAATGCGGTCAATGTTTTCTCGC-3', as shown in SEQ ID NO: 3;
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