Molecular Typing of Klebsiella Isolates

For PFGE, bacterial DNA was digested with XbaI (Fermentas, Milan, Italy) according to PulseNet protocol using conditions of pulse times from 5 to 40 sec over 24 hrs at 6.0V/cm and at 14°C.31 (link) Pulsotypes were analyzed through BioNumerics software (Applied Maths, Sint-Martens-Latem, Belgium), and dendrograms were generated using Dice coefficient and unweighted pair group method with arithmetic mean (UPGMA).9 (link) The similarity band patterns interpretation was performed according to Tenover criteria,24 (link),32 (link) setting 85% and 95% similarity cut-off for identifying similar restriction patterns and clusters, respectively. A validated MLST scheme was used,33 (link) and PCR products were sequenced by Sanger method (Eurofins Genomics) to identify allelic profiles and assign the Sequence Type (ST). The allelic combination was analysed on Pasteur platform (

http://bigsdb.pasteur.fr/perl/bigsdb/bigsdb.pl?db=pubmlst_klebsiella_seqdef_public

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Di Tella D., Tamburro M., Guerrizio G., Fanelli I., Sammarco M.L, & Ripabelli G. (2019). Molecular Epidemiological Insights into Colistin-Resistant and Carbapenemases-Producing Clinical Klebsiella pneumoniae Isolates. Infection and Drug Resistance, 12, 3783-3795.

Publication 2019

XbaI Sanger method

Allelic Bacterial dna Klebsiella Martens Pfge Pulse

Corresponding Organization : University of Molise

Top 5 similar protocols

Variable analysis

independent variables

Restriction enzyme used for digestion of bacterial DNA (XbaI)

dependent variables

Pulsotypes analyzed through BioNumerics software
Similarity band patterns interpreted according to Tenover criteria
Allelic profiles and Sequence Type (ST) identified through MLST scheme

control variables

Conditions of pulse times from 5 to 40 sec over 24 hrs at 6.0V/cm and at 14°C
Dice coefficient and unweighted pair group method with arithmetic mean (UPGMA) used for dendrogram generation

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!