Influence of polymerase brand on microarray-based spoligotyping in low concentrations of mycobacterial DNA

Stefan Monecke; Ines Engelmann; Ralf Ehricht

doi:10.1016/j.mcp.2015.01.003

Influence of polymerase brand on microarray-based spoligotyping in low concentrations of mycobacterial DNA

Mol Cell Probes. 2015 Apr;29(2):126-8. doi: 10.1016/j.mcp.2015.01.003. Epub 2015 Feb 3.

Authors

Stefan Monecke¹, Ines Engelmann², Ralf Ehricht²

Affiliations

¹ Institute for Medical Microbiology and Hygiene, Technische Universität Dresden, Dresden, Germany; Alere Technologies GmbH, Jena, Germany; Infectognostics Research Campus Jena, Jena, Germany. Electronic address: monecke@rocketmail.com.
² Alere Technologies GmbH, Jena, Germany; Infectognostics Research Campus Jena, Jena, Germany.

PMID: 25656919
DOI: 10.1016/j.mcp.2015.01.003

Abstract

Spoligotyping is a widely used typing method for the Mycobacterium tuberculosis complex. Protocols and platforms can be adapted for direct use on patient samples. Serial dilutions of genomic DNA from Mycobacterium bovis BCG strain DSM45071 were spoligotyped by array hybridization using 32 different commercial PCR polymerase preparations. In samples with very low concentrations of mycobacterial DNA, commercially available PCR polymerases differed in their performance, and some yielded no, or false, identification. Direct spoligotyping from samples with very low concentrations of mycobacterial DNA thus requires careful selection of polymerase and strict standardization.

Keywords: BCG; Mycobacterium tuberculosis complex; Polymerase; Spoligotyping.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Typing Techniques
DNA, Bacterial / genetics*
DNA-Directed DNA Polymerase / classification*
Humans
Mycobacterium tuberculosis / classification*
Mycobacterium tuberculosis / genetics

Substances

DNA, Bacterial
DNA-Directed DNA Polymerase