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Pyrosequencing as a method for grouping of Listeria monocytogenes strains on the basis of single-nucleotide polymorphisms in the inlB gene - PubMed

Pyrosequencing as a method for grouping of Listeria monocytogenes strains on the basis of single-nucleotide polymorphisms in the inlB gene

H Unnerstad et al. Appl Environ Microbiol. 2001 Nov.

Abstract

By using pyrosequencing (i.e., sequencing by synthesis) 106 strains of different serovars of Listeria monocytogenes were rapidly grouped into four categories based on nucleotide variations at positions 1575 and 1578 of the inlB gene. Strains of serovars 1/2a and 1/2c constituted one group, and strains of serovars 1/2b and 3b constituted another group, whereas serovar 4b strains were separated into two groups.

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Figures

FIG. 1
FIG. 1

Pyrograms obtained by pyrosequencing of L. monocytogenes strains belonging to different serovars, based on positions 1575 and 1578 of the inlB gene. Incorporated nucleotides are shown below each pyrogram. Relative light units are indicated on the ordinate axes. Additions of enzyme (E) and substrate (S) mixtures and nucleotides are indicated on the horizontal axes. (A) Serovar 1/2a strain SLU 1906 representing the serovar 1/2a-serovar 1/2c group as determined by T and A at the two variable positions. (B) Serovar 3b strain SLU 1182 representing the serovar 1/2b-serovar 3b group as determined by T and T at the two variable positions. (C) Serovar 4b strain SLU 249 representing the serovar 4b-I group as determined by C/T and G/T at the two variable positions. (D) Serovar 4b strain SLU 28 representing the serovar 4b-II group as determined by T and G/T at the two variable positions.

FIG. 2
FIG. 2

Southern blot hybridization with a 104-bp inlB probe of EcoRI-digested (A) and NcoI-digested (B) genomic DNAs from L. monocytogenes strains belonging to different serovars. Lane 1, serovar 1/2a strain SLU 83; lane 2, serovar 4b-I strain SLU 520; lane 3, serovar 1/2c strain SLU 524; lane 4, serovar 4b-II strain SLU 559; lane 5, serovar 3b strain SLU 594; lane 6, serovar 1/2b strain SLU 2324; lane M, λ DNA HindIII marker. Molecular sizes (in kilobases) are indicated on the left.

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