PFGE and MLST of K. pneumoniae

Post-extraction DNA was digested with 45U Xbal (Takara Biotech) for 2 hours at 37°C. We used CHEFDRIII apparatus (Bio-Rad Laboratories, Hercules, CA, USA) to perform PFGE for K. pneumoniae isolates as previously described.23 (link) PCR assay was performed to amplify internal portions of the seven housekeeping genes of K. pneumoniae (gapA, infB, mdh, pgi, phoE, rpoB, and tonB) with specific primers.24 (link) Amplified products were sequenced from private enterprise (Sangon Biotech-Shanghai, China

http://www.pasteur.fr/recherche/genopole/PF8/mlst/Kpneumoniae

) and were referred to assign sequence types (STs).

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Patil S., Chen H., Guo C., Zhang X., Ren P.G., Francisco N.M, & Wen F. (2021). Emergence of Klebsiella pneumoniae ST307 Co-Producing CTX-M with SHV and KPC from Paediatric Patients at Shenzhen Children’s Hospital, China. Infection and Drug Resistance, 14, 3581-3588.

Publication 2021

45U Xbal CHEFDRIII apparatus

Assay Housekeeping genes K pneumoniae Pfge Primers

Corresponding Organization : Shenzhen Children's Hospital

Other organizations : Shenzhen University, University Town of Shenzhen

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Variable analysis

independent variables

Xbal digestion (45U, 2 hours at 37°C)

dependent variables

PFGE patterns of K. pneumoniae isolates
Amplified products of internal portions of seven housekeeping genes (gapA, infB, mdh, pgi, phoE, rpoB, and tonB) in K. pneumoniae
Sequence types (STs) of K. pneumoniae isolates

control variables

CHEFDRIII apparatus (Bio-Rad Laboratories, Hercules, CA, USA) for PFGE
PCR assay with specific primers to amplify internal portions of the seven housekeeping genes
Sequencing of amplified products by private enterprise (Sangon Biotech-Shanghai, China)

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