Multilocus Sequence Typing of Salmonella

Genomic DNA was isolated using the InstaGene Matrix (Bio-Rad, Hercules, CA, USA) according to the manufacturer’s protocol. All primer sequences for amplification and sequencing were obtained from the MLST Databases of the University of Warwick (www.mlst.warwick.ac.uk/mlst/dbs/Senterica, accessed on 21 June 2023). The PCR cycling conditions were as indicated in instructions posted on the website. The PCR products were purified using Sephadex G-50 fine resin (GE Healthcare Bio-Sciences AB, Uppsala, Sweden). Nucleotide cycle-sequencing was performed directly on purified PCR templates using automated Sanger dideoxy chain termination methods and the primers described on the MLST website. Sequences of seven housekeeping genes (aroC, dnaN, hemD, hisD, purE, sucA, and thrA) were compared with those in the MLST database (http://mlst.warwick.ac.uk/mlst/dbs/Senterica accessed on 21 June 2023) to obtain the allele number and sequence type (ST) number for each isolate. Sequence information for newly assigned alleles and STs was deposited in the MLST database [17 (link)].

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Gao Y., Chen K., Lin R., Xu X., Xu F., Lin Q., Hu Y., Zhang H., Zhang J., Liao M, & Qu X. (2023). High Levels of Antibiotic Resistance in MDR-Strong Biofilm-Forming Salmonella Typhimurium ST34 in Southern China. Microorganisms, 11(8), 2005.

Publication 2023

InstaGene Matrix Sephadex G-50 fine resin

Alleles Genomic Housekeeping genes Primers Resin Sephadex g 50 Suca Thra

Corresponding Organization : Shanghai Municipal Center For Disease Control Prevention

Other organizations : Chinese Academy of Sciences, University of Macau

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Variable analysis

independent variables

Primer sequences for amplification and sequencing

dependent variables

Nucleotide sequences of seven housekeeping genes (aroC, dnaN, hemD, hisD, purE, sucA, and thrA)

control variables

Genomic DNA isolation using InstaGene Matrix
PCR cycling conditions as per instructions
PCR product purification using Sephadex G-50 fine resin
Nucleotide cycle-sequencing using automated Sanger dideoxy chain termination methods

controls

Positive control: Comparison of sequences with those in the MLST database to obtain allele number and sequence type (ST) number for each isolate
Negative control: Not explicitly mentioned

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