Genetic Relatedness of K. pneumoniae Isolates

We performed PFGE to check whether there is the presence of any clonal transmission within the hospital and MLST was applied to assess the genetic relatedness of the identified isolates. Post extraction, DNA was digested with 45U Xbal (Takara Biotech) for 2 h at 37 °C. We used CHEFDRIII apparatus (Bio-Rad Laboratories, Hercules, CA, USA) to perform PFGE for K. pneumoniae isolates as per earlier described [13 (link)]. Amplified fragments of seven housekeeping genes (gapA, infB, mdh, pgi, phoE, rpoB, and tonB) were sequenced to MLST analysis and MLST website MLST website-http://www.pasteur.fr/recherche/genopole/PF8/mlst/Kpneumoniae was referred to assign the STs.

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Patil S., Chen X, & Wen F. (2019). Exploring the phenotype and genotype of multi-drug resistant Klebsiella pneumoniae harbouring blaCTX-M group extended-spectrum β-lactamases recovered from paediatric clinical cases in Shenzhen, China. Annals of Clinical Microbiology and Antimicrobials, 18, 32.

Publication 2019

45U Xbal CHEFDRIII apparatus

Clonal Housekeeping genes K pneumoniae Pfge Sequenced analysis Transmission

Corresponding Organization :

Other organizations : Shenzhen Children's Hospital

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Variable analysis

independent variables

Xbal enzyme digestion (45U, 2 h at 37 °C)

dependent variables

Presence of clonal transmission within the hospital (assessed by PFGE)
Genetic relatedness of the identified isolates (assessed by MLST)

control variables

PFGE protocol (as per earlier described [13])
MLST protocol (amplification and sequencing of seven housekeeping genes)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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