Research Of Molecular Diagnostic To Sulfate-Reducing Bacteria Based On Nucleic Acid Amplification

Metallic material corrosion is influenced by microbial corrosion,sulfate-reducing bacteria(SRB)is a kind of microorganisms which could promote metal corrosion.Based on the establishment of anaerobic culture system for sulfate-reducing bacteria,two kinds of nucleic acid amplification technique,polymerase chain reaction(PCR)and recombinase polymerase amplification(RPA)were used to establish one or two methods for rapid detection of SRB strains under the condition of successful cultivation of SRB.Desulfovibrio fructosivorans(Df,strain No.3468)and Desulfovibrio vulgaris(Dv),Desulfovibrio caledoniensis(Dc)and 7 SRB strains were obtained from the Chinese Academy of Sciences,Institute of Marine Research,Chinese Academy of Sciences.11 pairs of primers were designed based on the sequences of 16 SrDNA,dissimilatory sulfate reductase(DSR),adenosine-5’-phosphosulfate reductase(APS)from SRB genome.First,the efficacy of the primers was verified by polymerase chain reaction(PCR)with standard strain Desulfovibrio caledoniensis(Dc)genome,and the specificity of the primers was verified by multiple SRB strains and Escherichia coli DH5α.Two pairs of primers are capable of specifically amplifying part of SRB strains.On the basis of two pairs of specific primers verified by PCR,the method of detecting SRB by recombinase polymerase amplification(RPA)was explored.Four pairs of primers designed for RPA were used fot the detection of SRB.After identification,a pair of primers based on RPA could be efficacy and specific under PCR reaction system,which provided an important reference for the future optimization of RPA reaction system and primer design.