Sequences of FTLSV with associated location of origin, collection dates, host and GenBank accession numbers.

<p>Sequences of FTLSV with associated location of origin, collection dates, host and GenBank accession numbers.</p

Bayesian coalescent analysis of FTLSV based on the S segment.

<p>Maximum clade credibility and Maximum likelihood tree is shown with posterior probability values and bootstrap value depicted at the nodes. Phylogenetic analysis was carried out using Mrbayes 3.2.1 and Mega software 5, respectively. Designations of the different FTLSV phylogroups (A,B,C,E,F clade) are as indicated, based on previously defined assignments. These sequences are described in more detail in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003237#pntd-0003237-t001" target="_blank">Table 1</a>. The sequences from Japan were included in this analysis.</p

Extent of geographic clustering, as indicated by AI and PS statistics.

<p>Extent of geographic clustering, as indicated by AI and PS statistics.</p

Maximum clade credibility tree based on L genes summarized for FTLSV were generated using the geospatial Bayesian analysis.

<p>Posterior clade probabilities for key nodes were shown. The colors of the branches correspond to their probable geographic location as calculated using the geospatial analysis.</p

Bayesian coalescent and Maximum likelihood (ML) analysis of FTLSV based on the M segment.

<p>Maximum clade credibility and Maximum likelihood tree is shown with posterior probability values and bootstrap value depicted at the nodes. Phylogenetic analysis was carried out using Mrbayes 3.2.1 and Mega software 5, respectively. Designations of the different FTLSV phylogroups (A,B,C,D,E,F clade) are as indicated, based on previously defined assignments. These sequences are described in more detail in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003237#pntd-0003237-t001" target="_blank">Table 1</a>. The sequences from Japan were included in this analysis.</p

Simultaneous Detection and Identification of Enteric Viruses by PCR-Mass Assay

<div><p>Simultaneous detection of enteric viruses that cause similar symptoms (e.g. hand, foot and mouth disease) is essential to the prevention of outbreaks and control of infections. In this study, a novel PCR-Mass assay combining multiplex polymerase chain reaction (PCR) with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was developed and used for simultaneous detection of eight distinct human enteric viruses. Enteric viral isolates and standard viral RNAs were examined to determine the sensitivity and specificity of the PCR-Mass assay. Clinical performance was evaluated with a total of 101 clinical specimens from patients suspected of having hand, foot and mouth disease (HFMD). The results were compared to those of previous analyses using real-time RT-PCR. The identification of specific viruses and clinical specimens shows that the PCR-Mass assay performed as well as or better than standard methods with respect to indicating the presence of multiplex pathogens in a single specimen.</p> </div

Simultaneous detection of clinical specimens.

<p>EV71-CMN was added as positive control. (A) Sample positive for EV71, ECHO, and HEV; (B) Sample positive for EV71, ECHO, and coxA16.</p

Mass spectra of different enteric viruses detected by PCR-Mass.

<p>(A) ASTRV; (B) coxA16; (C) ECHO; (D) EV71; (E) HEV; (F) NVG5; (G) PLV; (H) REV.</p

Comparison of the PCR-Mass assay with real-time RT-PCR and sequencing methods for detection of enterovirus 71 and coxsackievirus A16 in clinical specimens.

a<p>Sequencing validated the EV71 positive for two of the four samples but invalidated for other two samples.</p>b<p>One sample was confirmed as coxA16 negative by sequencing.</p>c<p>Three samples initially detected as EV71 negative by PCR-Mass were confirmed as negative by sequencing.</p>d<p>Six samples initially detected as coxA16 negative by PCR-Mass were confirmed as negative by sequencing.</p

Extension products of the PCR-Mass assay.

<p>Extension products of the PCR-Mass assay.</p