The estimation selection in 7 housekeeping gene using the non-synonymous to synonymous amino acid substitution ratio (dN/dS) in this study.

<p>^† d_N/d_S: The ratio of mean non-synonymous substitutions per non-synonymous site and mean synonymous substitutions per synonymous site.</p><p>The estimation selection in 7 housekeeping gene using the non-synonymous to synonymous amino acid substitution ratio (dN/dS) in this study.</p

Population genetic structure of <i>O</i>. <i>tsutsugamushi</i> in Laos and Thailand.

<p>(A) <i>O</i>. <i>tsutsugamushi</i> population in Laos (n = 74). (B) The comparison of <i>O</i>. <i>tsutsugamushi</i> population in Laos and those in Thailand (n = 89) using comparative e-BURST in which blue dot were founders; pink letter were STs found in both sites; green letter were the STs found only in Laos and black demonstrated the ST found only in Thailand.</p

The geographic map of 3 study sites in Laos (Vientiane, Luang Namtha and Salavan).

<p>The geographic map of 3 study sites in Laos (Vientiane, Luang Namtha and Salavan).</p

The phylogenetic tree of concatenated sequence of <i>O</i>. <i>tsutsugamushi</i> isolates from 74 Laos and 89 Thailand using neighbor-joining method with Bootstrap value of 3000.

<p>The color triangle demonstrated isolates from different sites: black represented Vientiane; blue represented Luang Namtha; pink represented Salavan; grey were from Thailand and open triangle were reference isolates.</p

List of 7 housekeeping gene and primer sequences used in this study.

<p>List of 7 housekeeping gene and primer sequences used in this study.</p

Population differentiation (F_ST) based on concatenated sequences.

<p>Populations were defined according to four strain sources; three from Laos (Vientiane, Luang Namtha and Salavan) characterised in the current study, and one population from Thailand (Udon Thani) characterised previously. The F_ST values are depicted as a dendrogram using the neighbour-joining algorithm.</p

Sequence type of 74 <i>O</i>. <i>tsutsugamushi</i> isolates from Laos.

<p>The strain name prefix TM indicates samples from Vientiane, LNT those from Luang Namtha, and SV those from Salavan.</p

<i>B</i>. <i>pseudomallei</i> and other <i>Burkholderia</i> species used in this study.

<p><i>B</i>. <i>pseudomallei</i> and other <i>Burkholderia</i> species used in this study.</p

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the identification of <i>Burkholderia pseudomallei</i> from Asia and Australia and differentiation between <i>Burkholderia</i> species

<div><p>Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used for rapid bacterial identification. Studies of <i>Burkholderia pseudomallei</i> identification have involved small isolate numbers drawn from a restricted geographic region. There is a need to expand the reference database and evaluate <i>B</i>. <i>pseudomallei</i> from a wider geographic distribution that more fully captures the extensive genetic diversity of this species. Here, we describe the evaluation of over 650 isolates. Main spectral profiles (MSP) for 26 isolates of <i>B</i>. <i>pseudomallei</i> (N = 5) and other <i>Burkholderia</i> species (N = 21) were added to the Biotyper database. MALDI-TOF MS was then performed on 581 <i>B</i>. <i>pseudomallei</i>, 19 <i>B</i>. <i>mallei</i>, 6 <i>B</i>. <i>thailandensis</i> and 23 isolates representing a range of other bacterial species. <i>B</i>. <i>pseudomallei</i> originated from northeast and east Thailand (N = 524), Laos (N = 12), Cambodia (N = 14), Hong Kong (N = 4) and Australia (N = 27). All 581 <i>B</i>. <i>pseudomallei</i> were correctly identified, with 100% sensitivity and specificity. Accurate identification required a minimum inoculum of 5 x 10⁷ CFU/ml, and identification could be performed on spiked blood cultures after 24 hours of incubation. Comparison between a dendrogram constructed from MALDI-TOF MS main spectrum profiles and a phylogenetic tree based on <i>recA</i> gene sequencing demonstrated that MALDI-TOF MS distinguished between <i>B</i>. <i>pseudomallei</i> and <i>B</i>. <i>mallei</i>, while the <i>recA</i> tree did not. MALDI-TOF MS is an accurate method for the identification of <i>B</i>. <i>pseudomallei</i>, and discriminates between this and other related <i>Burkholderia</i> species.</p></div

Dissociation curves of the EV-A71 SYBR Green RT-PCR performed on dilutions of plasmid B1.

<p>Dissociation curves of the EV-A71 SYBR Green RT-PCR performed on dilutions of plasmid B1.</p